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Organoids | September 2024 - Browse Articles

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<a href="https://doi.org/10.3390/organoids3030014">https://doi.org/10.3390/organoids3030014</a> - 4 Sep 2024 </div> Viewed by 1619 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Recent advancements in vascular organoid (VO) and vessel-on-chip (VoC) technologies have revolutionized our approach to studying human diseases, offering unprecedented insights through more physiologically relevant models. VOs generated from human pluripotent stem cells exhibit remarkable self-organization capabilities, forming complex three-dimensional structures that closely <a href="#" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/14/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Recent advancements in vascular organoid (VO) and vessel-on-chip (VoC) technologies have revolutionized our approach to studying human diseases, offering unprecedented insights through more physiologically relevant models. VOs generated from human pluripotent stem cells exhibit remarkable self-organization capabilities, forming complex three-dimensional structures that closely mimic human blood vessel architecture and function, while VoCs are engineered with microfluidic systems that meticulously recreate the physical and functional attributes of blood vessels. These innovative constructs serve as powerful tools for investigating vascular development, disease progression, and therapeutic efficacy. By enabling the creation of patient-specific VOs and VoCs, they pave the way for personalized medicine approaches, allowing researchers to delve into genetic variations, intricate cellular interactions, and dynamic processes with exceptional resolution. The synergy between VOs and VoCs with newly developed cutting-edge technologies has further amplified their potential, unveiling novel mechanisms underlying human pathologies and identifying promising therapeutic targets. Herein, we summarize different types of VOs and VoCs and present an extensive overview on the generation and applications of VOs and VoCs. We will also highlight clinical and translational challenges and future perspectives around VOs and VoCs. <a href="/2674-1172/3/3/14">Full article</a> </div> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/14/show" ><span >&#9658;</span><span style=" display: none;">&#9660;</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1471095"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1471095"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1471095" data-cycle-prev="#prev1471095" data-cycle-progressive="#images1471095" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1471095-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g001-550.jpg?1725440574" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1471095" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1471095-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g002-550.jpg?1725440577'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1471095-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g003-550.jpg?1725440579'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1471095-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g004-550.jpg?1725440580'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1471095-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g005-550.jpg?1725440582'><p>Figure 5</p></div></script></div></div><div id="article-1471095-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g001-550.jpg?1725440574" title=" <strong>Figure 1</strong><br/> &lt;p&gt;&lt;b&gt;Diagram showing the main methods to generate vascular organoids (VOs), Created with &lt;a href=&quot;http://BioRender.com&quot; target=&quot;_blank&quot;&gt;BioRender.com&lt;/a&gt;&lt;/b&gt;. The basic protocol for VO generation from hPSCs to hiPSCs is based on a stepwise differentiation of hPSC (or hiPSC) aggregates with sequential changing of culture media and further differentiation into vascular networks in a 3D matrix, with a variety of modifications as reported by Wimmer et al. [&lt;a href=&quot;#B15-organoids-03-00014&quot; class=&quot;html-bibr&quot;&gt;15&lt;/a&gt;], Schmidt et al. [&lt;a href=&quot;#B16-organoids-03-00014&quot; class=&quot;html-bibr&quot;&gt;16&lt;/a&gt;], and Dailamy et al. [&lt;a href=&quot;#B17-organoids-03-00014&quot; class=&quot;html-bibr&quot;&gt;17&lt;/a&gt;], respectively. It is worth noting that Penninger’s group [&lt;a href=&quot;#B15-organoids-03-00014&quot; class=&quot;html-bibr&quot;&gt;15&lt;/a&gt;,&lt;a href=&quot;#B18-organoids-03-00014&quot; class=&quot;html-bibr&quot;&gt;18&lt;/a&gt;,&lt;a href=&quot;#B19-organoids-03-00014&quot; class=&quot;html-bibr&quot;&gt;19&lt;/a&gt;] reported the first generation of VOs from hPSCs, which was quickly adapted by other laboratories such as Romeo et al. [&lt;a href=&quot;#B20-organoids-03-00014&quot; class=&quot;html-bibr&quot;&gt;20&lt;/a&gt;] and Nikolova et al. [&lt;a href=&quot;#B21-organoids-03-00014&quot; class=&quot;html-bibr&quot;&gt;21&lt;/a&gt;]. MIM, mesoderm induction medium (DMEM-F12, 2 mM, Ascorbic acid, 355 μM CHIR 99021, 10 μM BMP4); VGM, vascular growth medium (Neurobasal medium/DMEM-F12, N2B27, 2 mM Ascorbic acid, 100 ng/mL VEGF-A); OMM, organoid maintenance medium (Neurobasal medium/DMEM-F12, N2B27, 2 mM Ascorbic acid); VIM, vascular induction media (N2/B27 + VEGF + FSK); VMM, vascular maturation media (StemPro + 15%FBS + VEGF + bFGF).&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/14'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g002-550.jpg?1725440577" title=" <strong>Figure 2</strong><br/> &lt;p&gt;&lt;b&gt;Schematic illustration of the key applications for vascular organoids (VOs)&lt;/b&gt;. Increasing numbers of studies demonstrate a variety of promising applications for hPSC-derived VOs, including infectious disease pathogenesis (such as SARS-CoV2 and pathogenic bacteria), in vitro vascular disease modeling, high-throughput drug screening and drug toxicity testing, creating human VO biobanks for regenerative medicine, multi-omics analysis to probe novel insights into signaling pathways underlying vascular development and disease etiology, exploring key developmental events in human vascular systems, genetic engineering and editing for inherited disorders, and personalized and precision medicine. Diagram is created with &lt;a href=&quot;http://BioRender.com&quot; target=&quot;_blank&quot;&gt;BioRender.com&lt;/a&gt;.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/14'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g003-550.jpg?1725440579" title=" <strong>Figure 3</strong><br/> &lt;p&gt;Schematic diagram illustrating the main strategies to generate vessel-on-chip (VoC) and potential applications of VoCs. Different functional vascular cells can be derived from human-induced pluripotent stem cells (hiPSCs), which are incorporated into various 3D microporous scaffolds to create functional VoCs using a variety of fabrication techniques and microfluidic strategies. These human VoCs offer a wide range of potential applications, encompassing various fields such as studying vascular physiology, pathology, and potential therapeutic interventions, exploring infectious disease pathogenesis, using for in vitro vascular disease modeling as well as high-throughput drug screening and drug toxicity testing, providing novel insights into vascular aging and angiogenesis, and creating human VoC biobanks for regenerative medicine. EC, endothelial cell; SMC, smooth muscle cells; SMART, substrate modification and replication by thermoforming. Diagram is created with &lt;a href=&quot;http://BioRender.com&quot; target=&quot;_blank&quot;&gt;BioRender.com&lt;/a&gt;.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/14'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g004-550.jpg?1725440580" title=" <strong>Figure 4</strong><br/> &lt;p&gt;&lt;b&gt;Schematic diagram illustrating the main strategies to generate vascularized organoids&lt;/b&gt;. Potentially, multiple methods could be applied to generate vascularized tissue organoids such as in vivo transplantation of tissues-specific organoids into mice which allows for host vasculature integration, co-culturing either hiPSCs/hPSCs with mature endothelial cells or hPSC-derived tissues-specific cells with hPSC-derived endothelial cells, genetic engineering vascular-inducing transcription factors (TFs, such as ETV2) into tissue-specific organoids, including vascular induction factors (such as VEGF-A, WNTs, BMPs) into tissue-specific organoids, and different fusion strategies (co-incubating tissue-specific spheroids/EBs (embryonic body) with vascular spheroids/EBs with our without ECM scaffolds, or co-culturing tissue-specific organoids with vascular organoids (Vos)). Diagram is created with &lt;a href=&quot;http://BioRender.com&quot; target=&quot;_blank&quot;&gt;BioRender.com&lt;/a&gt;.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/14'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00014/article_deploy/html/images/organoids-03-00014-g005-550.jpg?1725440582" title=" <strong>Figure 5</strong><br/> &lt;p&gt;&lt;b&gt;Schematic diagram illustrating human multiple organs-on-chip (MOoC)&lt;/b&gt;. HiPSCs/hPSCs-derived tissue-specific cells or organoids could be integrated into a human MOoC system by using sophisticated microfluidic devices or systems. These MOoC systems recapitulate the complex physiological environments and interactions of multiple organ systems and better mimic humans, allowing for the study of complex physiological system-system interactions and diseases in a high-throughput controlled manner. BOs, brain organoids; COs, cardiac organoids; IOs, intestinal organoids; KOs, kidney organoids; LOs, lung organoids; POs, pancreatic organoids; VOs, vascular organoids. Diagram is created with &lt;a href=&quot;http://BioRender.com&quot; target=&quot;_blank&quot;&gt;BioRender.com&lt;/a&gt;.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/14'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1461011" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <a data-dropdown="drop-supplementary-1461011" aria-controls="drop-supplementary-1461011" aria-expanded="false" title="Supplementary Material"> <i class="material-icons">attachment</i> </a> <div id="drop-supplementary-1461011" class="f-dropdown label__btn__dropdown label__btn__dropdown--wide" data-dropdown-content aria-hidden="true" tabindex="-1"> Supplementary material: <br/> <a href="/2674-1172/3/3/13/s1?version=1724248447"> Supplementary File 1 (ZIP, 503 KiB) </a><br/> </div> </div> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 9 pages, 5929 KiB &nbsp; </span> <a href="/2674-1172/3/3/13/pdf?version=1724248446" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="A Method to Study Migration and Invasion of Mouse Intestinal Organoids" data-journal="organoids"> <i class="material-icons custom-download"></i> </a> </div> <div class="article-icons"><span class="label openaccess" data-dropdown="drop-article-label-openaccess" aria-expanded="false">Open Access</span><span class="label articletype">Study Protocol</span></div> <a class="title-link" href="/2674-1172/3/3/13">A Method to Study Migration and Invasion of Mouse Intestinal Organoids</a> <div class="authors"> by <span class="inlineblock "><strong>Valérie M. Wouters</strong>, </span><span class="inlineblock "><strong>Ciro Longobardi</strong> and </span><span class="inlineblock "><strong>Jan Paul Medema</strong></span> </div> <div class="color-grey-dark"> <em>Organoids</em> <b>2024</b>, <em>3</em>(3), 194-202; <a href="https://doi.org/10.3390/organoids3030013">https://doi.org/10.3390/organoids3030013</a> - 21 Aug 2024 </div> Viewed by 897 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Colorectal cancer (CRC) is the third most common cancer worldwide and it is the second leading cause of cancer death. In CRC, as in most cancers, the formation of metastasis through the migration and invasion of cancer cells to distant organs is associated <a href="#" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/13/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Colorectal cancer (CRC) is the third most common cancer worldwide and it is the second leading cause of cancer death. In CRC, as in most cancers, the formation of metastasis through the migration and invasion of cancer cells to distant organs is associated with a dismal prognosis. The study of the mechanisms associated with cancer, and, in particular, CRC, changed in the last decade due to the introduction of organoids. These represent a step forward in terms of complexity from cell lines and allowed the use of mouse models in cancer research to be limited. Although organoids faithfully model the cellular complexity of CRC, current protocols do not allow for the use of organoids in some crucial processes of metastasis, such as migration and invasion. In this study, a method to study migration and invasion using mouse intestinal organoids in vitro is presented. This protocol provides researchers with the opportunity to investigate the migratory behavior of organoid lines and study the impact of distinct mutations on the migratory and invasive capacity of cancer cells. <a href="/2674-1172/3/3/13">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Special Issue <a href=" /journal/organoids/special_issues/5JAANIW6XT ">Organoids and Cancer Models</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/13/show" ><span >&#9658;</span><span style=" display: none;">&#9660;</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1461011"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1461011"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1461011" data-cycle-prev="#prev1461011" data-cycle-progressive="#images1461011" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1461011-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/organoids/organoids-03-00013/article_deploy/html/images/organoids-03-00013-g001-550.jpg?1724248570" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1461011" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1461011-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00013/article_deploy/html/images/organoids-03-00013-g002-550.jpg?1724248571'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1461011-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00013/article_deploy/html/images/organoids-03-00013-g003-550.jpg?1724248573'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1461011-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00013/article_deploy/html/images/organoids-03-00013-g004-550.jpg?1724248575'><p>Figure 4</p></div></script></div></div><div id="article-1461011-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/organoids/organoids-03-00013/article_deploy/html/images/organoids-03-00013-g001-550.jpg?1724248570" title=" <strong>Figure 1</strong><br/> &lt;p&gt;Schematic representation of the organoid transwell migration assay.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/13'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00013/article_deploy/html/images/organoids-03-00013-g002-550.jpg?1724248571" title=" <strong>Figure 2</strong><br/> &lt;p&gt;Optimization of transwell migration assay with gradient dependent migration. (&lt;b&gt;A&lt;/b&gt;). Schematic representation of cell lines transwell migration assay. (&lt;b&gt;B&lt;/b&gt;) Schematic representation of organoid transwell migration assay using different chemoattractants placed in the lower chambers. (&lt;b&gt;C&lt;/b&gt;) Quantification of KPN organoid transwell migration assay. Bar plots display the percentage of area of the membrane covered by cells. ns = not significant, ** &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.01 in paired &lt;span class=&quot;html-italic&quot;&gt;t&lt;/span&gt;-test. (&lt;b&gt;D&lt;/b&gt;) Representative images of KPN organoid transwell migration assay using different chemoattractants placed in the lower chambers. Scale bar 330 µm.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/13'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00013/article_deploy/html/images/organoids-03-00013-g003-550.jpg?1724248573" title=" <strong>Figure 3</strong><br/> &lt;p&gt;Comparison of migration between different organoid models. (&lt;b&gt;A&lt;/b&gt;) Representative images of A, K, KP and KPN organoid transwell migration assay. Scale bar 330 µm. (&lt;b&gt;B&lt;/b&gt;) Quantification of A, K, KP and KPN organoid transwell migration assay. &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.01 ordinary one-way ANOVA test.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/13'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00013/article_deploy/html/images/organoids-03-00013-g004-550.jpg?1724248575" title=" <strong>Figure 4</strong><br/> &lt;p&gt;Optimization of transwell invasion assay with Matrigel-coated membrane. (&lt;b&gt;A&lt;/b&gt;) Schematic representation of organoid transwell invasion assay. (&lt;b&gt;B&lt;/b&gt;) Representative images of KPN organoid transwell invasion assay 3 and 4 days after seeding. Scale bar 330 µm. (&lt;b&gt;C&lt;/b&gt;) Quantification of KPN organoid transwell invasion assay 0, 3 and 4 days after seeding. ** &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.01 in Welch’s &lt;span class=&quot;html-italic&quot;&gt;t&lt;/span&gt;-test.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/13'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1448404" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 20 pages, 1328 KiB &nbsp; </span> <a href="/2674-1172/3/3/12/pdf?version=1722580762" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Trophoblast Organoids: Capturing the Complexity of Early Placental Development In Vitro" data-journal="organoids"> <i class="material-icons custom-download"></i> </a> </div> <div class="article-icons"><span class="label openaccess" data-dropdown="drop-article-label-openaccess" aria-expanded="false">Open Access</span><span class="label articletype">Review</span></div> <a class="title-link" href="/2674-1172/3/3/12">Trophoblast Organoids: Capturing the Complexity of Early Placental Development In Vitro</a> <div class="authors"> by <span class="inlineblock "><strong>Brady M. Wessel</strong>, </span><span class="inlineblock "><strong>Jenna N. Castro</strong> and </span><span class="inlineblock "><strong>Victoria H. J. Roberts</strong></span> </div> <div class="color-grey-dark"> <em>Organoids</em> <b>2024</b>, <em>3</em>(3), 174-193; <a href="https://doi.org/10.3390/organoids3030012">https://doi.org/10.3390/organoids3030012</a> - 2 Aug 2024 </div> Viewed by 1282 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> First trimester placental development comprises some of the most critical yet understudied events that impact fetal development. Improper placentation leads to a host of health issues that not only impact the fetal period but also influence offspring throughout their lives. Thus, a paradigm <a href="#" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/12/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> First trimester placental development comprises some of the most critical yet understudied events that impact fetal development. Improper placentation leads to a host of health issues that not only impact the fetal period but also influence offspring throughout their lives. Thus, a paradigm to study early placental development is necessary, and this has spurred on the pursuit of new in vitro model systems that recapitulate specific aspects of placentation. One of the most complex and translationally valid models to arise are organoids, three-dimensional structures comprising multiple differentiated cell types that originate from a common progenitor population. Trophoblasts are the progenitor cells of the placenta, serving as the proliferative base for placental development. Recent advances have enabled the derivation of organoids from primary tissue, yet access to first trimester human samples is ethically constrained; derivation from established trophoblast stem cell lines is an alternative source. Organoids have already proven useful in generating insights into molecular events that underlie trophoblast differentiation, with the identification of new cell subtypes that are primed to differentiate down different paths. In this review, (1) we recap early pregnancy development events, (2) provide an overview of the cellular complexity of the placenta, (3) discuss the generation of organoids from tissue versus cellular sources, (4) highlight the value of translational animal models, and (5) focus on the complexities of the molecular regulation of trophoblast organoid development, differentiation, and function. <a href="/2674-1172/3/3/12">Full article</a> </div> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/12/show" ><span >&#9658;</span><span style=" display: none;">&#9660;</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1448404"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1448404"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1448404" data-cycle-prev="#prev1448404" data-cycle-progressive="#images1448404" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1448404-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/organoids/organoids-03-00012/article_deploy/html/images/organoids-03-00012-ag-550.jpg?1722835484" alt="" style="border: 0;"><p>Graphical abstract</p></div><script id="images1448404" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1448404-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00012/article_deploy/html/images/organoids-03-00012-g001-550.jpg?1722580912'><p>Figure 1</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1448404-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00012/article_deploy/html/images/organoids-03-00012-g002-550.jpg?1722580913'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1448404-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00012/article_deploy/html/images/organoids-03-00012-g003-550.jpg?1722580915'><p>Figure 3</p></div></script></div></div><div id="article-1448404-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/organoids/organoids-03-00012/article_deploy/html/images/organoids-03-00012-ag-550.jpg?1722835484" title=" <strong>Graphical abstract</strong><br/><strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/12'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00012/article_deploy/html/images/organoids-03-00012-g001-550.jpg?1722580912" title=" <strong>Figure 1</strong><br/> &lt;p&gt;First trimester trophoblast cell phenotypes. Schematic representation of the tip of an anchoring villous at the maternal/fetal interface. The fetal side is noted in white, while maternal decidua is noted in pink. Cellular anatomy is noted with the spatial distributions of all defined trophoblast cells shown.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/12'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00012/article_deploy/html/images/organoids-03-00012-g002-550.jpg?1722580913" title=" <strong>Figure 2</strong><br/> &lt;p&gt;Overview of trophoblast organoid sources. Routes of access to trophoblast organoids. The trophoblast stem cell route has three starting points, all of which converge on TSC culture in 2D. Primary cell isolations from the placenta can be obtained from first trimester (&lt;b&gt;middle&lt;/b&gt;) and term (&lt;b&gt;right&lt;/b&gt;) samples. These tissues are both digested in a similar fashion to yield proliferative cytotrophoblast cells. All three routes converge when proliferative cytotrophoblast cells are placed into a 3D paradigm, yielding trophoblast organoids.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/12'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00012/article_deploy/html/images/organoids-03-00012-g003-550.jpg?1722580915" title=" <strong>Figure 3</strong><br/> &lt;p&gt;Signaling pathways in trophoblast organoids. A broad overview of key signaling pathways that are known to govern trophoblast differentiation in TOs. Media-manipulated pathways are shown with solid lines, whereas indirect/autocrine-activated signaling is shown with dashed lines. The contributions of TO media EVT#1 and EVT#2 (differing growth factor compositions) are shown with arrows. Inhibition of Wnt drives EVT differentiation. TGF-β is continuously inhibited in this model but may not always be.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/12'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1447515" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 9 pages, 1065 KiB &nbsp; </span> <a href="/2674-1172/3/3/11/pdf?version=1722491457" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Organoids and 3D In Vitro Models as a Platform for Precision Medicine (PM): An Update" data-journal="organoids"> <i class="material-icons custom-download"></i> </a> </div> <div class="article-icons"><span class="label openaccess" data-dropdown="drop-article-label-openaccess" aria-expanded="false">Open Access</span><span class="label articletype">Opinion</span></div> <a class="title-link" href="/2674-1172/3/3/11">Organoids and 3D In Vitro Models as a Platform for Precision Medicine (PM): An Update</a> <div class="authors"> by <span class="inlineblock "><strong>Payal Ganguly</strong></span> </div> <div class="color-grey-dark"> <em>Organoids</em> <b>2024</b>, <em>3</em>(3), 165-173; <a href="https://doi.org/10.3390/organoids3030011">https://doi.org/10.3390/organoids3030011</a> - 1 Aug 2024 </div> Viewed by 1143 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> Globally, a number of diseases impact us and while treatment options exist, it is often found that similar treatments have variable effects on different patients with the same disease. Particularly in the case of conditions that are closely associated with genetics (like cancer), <a href="#" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/11/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> Globally, a number of diseases impact us and while treatment options exist, it is often found that similar treatments have variable effects on different patients with the same disease. Particularly in the case of conditions that are closely associated with genetics (like cancer), the intensity and results of a treatment vary between patients. Even for diseases like arthritis it is not uncommon for only a fraction of patients to achieve remission with the same therapeutic approach. With millions suffering from diseases like cancer and arthritis, precision medicine (PM) has been at the forefront of biomedical and pharmaceutical research since 2015. PM focusses on understanding the genetic and environmental factors affecting the patients and has several platforms. One of the platforms is the use of three-dimensional (3D) in vitro models, especially those derived from the patient themselves. These models, like organ-on-chip (OOC), organoid and spheroid models, 3D biomaterial scaffolds and others, have several advantages over traditional two-dimensional (2D) cell culture approaches. In this opinion paper, the author briefly discusses the different platforms used for PM. Then, the advantages that 3D in vitro models have over traditional 2D models and in vivo models are considered and an overview of their applications is provided. Finally, the author outlines the challenges and future directions and shares their opinion about using 3D in vitro models as a tool for PM towards enhanced patient outcomes. <a href="/2674-1172/3/3/11">Full article</a> </div> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/11/show" ><span >&#9658;</span><span style=" display: none;">&#9660;</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1447515"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1447515"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1447515" data-cycle-prev="#prev1447515" data-cycle-progressive="#images1447515" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1447515-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/organoids/organoids-03-00011/article_deploy/html/images/organoids-03-00011-g001-550.jpg?1722491531" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1447515" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1447515-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00011/article_deploy/html/images/organoids-03-00011-g002-550.jpg?1722491532'><p>Figure 2</p></div></script></div></div><div id="article-1447515-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/organoids/organoids-03-00011/article_deploy/html/images/organoids-03-00011-g001-550.jpg?1722491531" title=" <strong>Figure 1</strong><br/> &lt;p&gt;Platforms for precision medicine.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/11'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00011/article_deploy/html/images/organoids-03-00011-g002-550.jpg?1722491532" title=" <strong>Figure 2</strong><br/> &lt;p&gt;Applications of organoids and 3D models in PM.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/11'>Full article</a></strong> "></a></div> </div> </div> <div class="generic-item article-item"> <input class="article-list-checkbox export-element" type="checkbox" name="articles_ids[]" value="1429307" data-select-all-name="article-listing"> <div class="article-content"> <div class="label right label__btn"> <a data-dropdown="drop-supplementary-1429307" aria-controls="drop-supplementary-1429307" aria-expanded="false" title="Supplementary Material"> <i class="material-icons">attachment</i> </a> <div id="drop-supplementary-1429307" class="f-dropdown label__btn__dropdown label__btn__dropdown--wide" data-dropdown-content aria-hidden="true" tabindex="-1"> Supplementary material: <br/> <a href="/2674-1172/3/3/10/s1?version=1720162364"> Supplementary File 1 (ZIP, 163 KiB) </a><br/> </div> </div> <div class="label right label__btn"> <span style="font-size: 12px; color: #1a1a1a;"> 17 pages, 7889 KiB &nbsp; </span> <a href="/2674-1172/3/3/10/pdf?version=1720751385" class="UD_Listings_ArticlePDF" title="Article PDF" data-name="Heparin-Binding Epidermal-like Growth Factor (HB-EGF) Reduces Cell Death in an Organoid Model of Retinal Damage" data-journal="organoids"> <i class="material-icons custom-download"></i> </a> </div> <div class="article-icons"><span class="label openaccess" data-dropdown="drop-article-label-openaccess" aria-expanded="false">Open Access</span><span class="label articletype">Article</span></div> <a class="title-link" href="/2674-1172/3/3/10">Heparin-Binding Epidermal-like Growth Factor (HB-EGF) Reduces Cell Death in an Organoid Model of Retinal Damage</a> <div class="authors"> by <span class="inlineblock "><strong>Michelle N. H. Tang</strong>, </span><span class="inlineblock "><strong>Mariya Moosajee</strong>, </span><span class="inlineblock "><strong>Najam A. Sharif</strong>, </span><span class="inlineblock "><strong>G. Astrid Limb</strong> and </span><span class="inlineblock "><strong>Karen Eastlake</strong></span> </div> <div class="color-grey-dark"> <em>Organoids</em> <b>2024</b>, <em>3</em>(3), 148-164; <a href="https://doi.org/10.3390/organoids3030010">https://doi.org/10.3390/organoids3030010</a> - 5 Jul 2024 </div> Viewed by 3675 <div class="abstract-div"> <a href="#" onclick="$(this).next('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> <strong>Abstract </strong> </a> <div class="abstract-cropped inline"> In zebrafish and various mammalian species, HB-EGF has been shown to promote M&uuml;ller glia proliferation and activation of repair mechanisms that have not been fully investigated in human retina. In the current study, 70- to 90-day-old human retinal organoids were treated with 20 <a href="#" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/10/more" onclick="$(this).parents('.abstract-cropped').toggleClass('inline').next('.abstract-full').toggleClass('inline'); return false;"> [...] Read more.</a> </div> <div class="abstract-full "> In zebrafish and various mammalian species, HB-EGF has been shown to promote M&uuml;ller glia proliferation and activation of repair mechanisms that have not been fully investigated in human retina. In the current study, 70- to 90-day-old human retinal organoids were treated with 20 &mu;M 4-hydroxytamoxifen (4-OHT), and CRX, REC, NRL, PAX6, VIM, GFAP, and VSX2 gene and protein expression were assessed at various times points after treatment. Organoids with or without 4-OHT-induced damage were then cultured with HB-EGF for 7 days. We showed that 20 &mu;M 4-OHT caused a reduction in the number of recoverin-positive cells; an increase in the number of TUNEL-positive cells; and downregulation of the photoreceptor gene markers CRX, NRL, and REC. Culture of organoids with HB-EGF for 7 days after 4-OHT-induced damage caused a marked reduction in the number of TUNEL-positive cells and small increases in the number of Ki67-positive cells and PAX6 and NOTCH1 gene expression. The current results suggest that treatment of human ESC-derived retinal organoids with 4-OHT may be used as a model of retinal degeneration in vitro. Furthermore, HB-EGF treatment of human retinal organoids increases proliferating M&uuml;ller cells, but only after 4-OHT induced damage, and may be an indication of Muller reactivity in response to photoreceptor damage. Further studies will aim to identify factors that may induce M&uuml;ller cell-mediated regeneration of the human retina, aiding in the development of therapies for retinal degeneration. <a href="/2674-1172/3/3/10">Full article</a> </div> </div> <div class="belongsTo" style="margin-bottom: 10px;"> (This article belongs to the Special Issue <a href=" /journal/organoids/special_issues/BHTOZ2892T ">The Current Applications and Potential of Stem Cell-Derived Organoids</a>)<br/> </div> <a href="#" class="abstract-figures-show" data-counterslink = "https://www.mdpi.com/2674-1172/3/3/10/show" ><span >&#9658;</span><span style=" display: none;">&#9660;</span> Show Figures </a><div class="abstract-image-preview "><div class="arrow left-arrow" id="prev1429307"><i class="fa fa-caret-left"></i></div><div class="arrow right-arrow" id="next1429307"><i class="fa fa-caret-right"></i></div><div class="absgraph cycle-slideshow manual" data-cycle-fx="scrollHorz" data-cycle-timeout="0" data-cycle-next="#next1429307" data-cycle-prev="#prev1429307" data-cycle-progressive="#images1429307" data-cycle-slides=">div" data-cycle-log="false"><div class='openpopupgallery cycle-slide' data-imgindex='0' data-target='article-1429307-popup'><span class="helper"></span><img src="data:image/gif;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g001-550.jpg?1720751458" alt="" style="border: 0;"><p>Figure 1</p></div><script id="images1429307" type="text/cycle" data-cycle-split="---"><div class='openpopupgallery' data-imgindex='1' data-target='article-1429307-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g002-550.jpg?1720751463'><p>Figure 2</p></div> --- <div class='openpopupgallery' data-imgindex='2' data-target='article-1429307-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g003-550.jpg?1720751466'><p>Figure 3</p></div> --- <div class='openpopupgallery' data-imgindex='3' data-target='article-1429307-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g004-550.jpg?1720751470'><p>Figure 4</p></div> --- <div class='openpopupgallery' data-imgindex='4' data-target='article-1429307-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g005-550.jpg?1720751473'><p>Figure 5</p></div> --- <div class='openpopupgallery' data-imgindex='5' data-target='article-1429307-popup'><span class="helper"></span><img src='https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g006-550.jpg?1720751474'><p>Figure 6</p></div></script></div></div><div id="article-1429307-popup" class="popupgallery" style="display: inline; line-height: 200%"><a href="https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g001-550.jpg?1720751458" title=" <strong>Figure 1</strong><br/> &lt;p&gt;Morphological changes induced by 4-OHT in retinal organoids. (&lt;b&gt;A&lt;/b&gt;) Phase contrast images of retinal organoids after 24 h treatment (black arrows) with or without the addition of different 4-OHT concentrations (10, 20, or 50 µM). Scale 200 μm. (blue arrows show bulging and loss of transparency) (&lt;b&gt;B&lt;/b&gt;) Representative confocal images of retinal organoid cross-sections showing TUNEL-positive nuclei (green) and counterstained with DAPI (blue) to indicate total cell nuclei. Scale 50 µm. ONL = outer nuclear layer; INL = inner nuclear layer. (&lt;b&gt;C&lt;/b&gt;) Box plot shows the average thickness (µm) of retinal layers in organoids after treatment with or without 4-OHT (10 or 20 µM) over a 7-day period (&lt;span class=&quot;html-italic&quot;&gt;n&lt;/span&gt; = 6). (&lt;b&gt;D&lt;/b&gt;) Box plot shows the percentage of TUNEL-positive cells in retinal organoids treated with 4-OHT (10 or 20 µM) as compared to those without treatment (control) over 7 days (&lt;span class=&quot;html-italic&quot;&gt;n&lt;/span&gt; = 3). * &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.05; ** &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.01; *** &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.001; Student’s &lt;span class=&quot;html-italic&quot;&gt;t&lt;/span&gt; test. One-way ANOVA. Error bars represent SEM.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/10'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g002-550.jpg?1720751463" title=" <strong>Figure 2</strong><br/> &lt;p&gt;Photoreceptor degeneration induced by 4-OHT. (&lt;b&gt;A&lt;/b&gt;) Representative confocal images of control retinal organoids and retinal organoids after 20 µM 4-OHT treatment for 24 h, 3 days, and 7 days. Sections show expression of recoverin (red) and cell death observed with TUNEL (green) staining. Cell nuclei were detected with DAPI (blue). (White arrows show rounded cells after damage) 40× magnification; scale 50 µm. ONL = outer nuclear layer; INL = inner nuclear layer. (&lt;b&gt;B&lt;/b&gt;) Scatter plot shows the number of recoverin-positive cells (per mm&lt;sup&gt;2&lt;/sup&gt;) in retinal organoids treated with 4-OHT (20 µM) as compared to those without treatment (control) at each time point (24 h, 3 days, 7 days). &lt;span class=&quot;html-italic&quot;&gt;n&lt;/span&gt; = 3; (each n represents 3 images). (&lt;b&gt;C&lt;/b&gt;) Graphs show the relative log2-fold change (FC) in expression of the photoreceptor-related genes CRX, NRL, and REC after 4-OHT treatment for 24 h, 3 days, or 7 days as compared to untreated control organoids. * &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.05; ** &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.01; &lt;span class=&quot;html-italic&quot;&gt;n&lt;/span&gt; = 4. Error bars represent SEM.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/10'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g003-550.jpg?1720751466" title=" <strong>Figure 3</strong><br/> &lt;p&gt;Müller cell responses to 4-OHT treatment. (&lt;b&gt;A&lt;/b&gt;,&lt;b&gt;E&lt;/b&gt;) Representative confocal images of retinal organoids after 20 µM 4-OHT treatment for 24 h, 3 days, and 7 days. Sections show expression of (&lt;b&gt;A&lt;/b&gt;) vimentin (green), Ki67 (red), and VSX2 (magenta) as well as (&lt;b&gt;E&lt;/b&gt;) vimentin (green), GFAP (red), and SOX2 (magenta). Cell nuclei were detected with DAPI (blue). 40× magnification; scale 50 µm. ONL = outer nuclear layer; INL = inner nuclear layer. (&lt;b&gt;B&lt;/b&gt;–&lt;b&gt;D&lt;/b&gt;) Dot plot shows the proportion of cells positive for Ki67 (&lt;b&gt;B&lt;/b&gt;) and VSX2 (Chx10) (&lt;b&gt;C&lt;/b&gt;) (per mm&lt;sup&gt;2&lt;/sup&gt;) and those that were double positive for Ki67 and VSX2 (per mm&lt;sup&gt;2&lt;/sup&gt;) (&lt;b&gt;D&lt;/b&gt;) in retinal organoids treated with 4-OHT (20 µM) as compared to those without treatment (control) at each time point (24 h, 3 days, and 7 days). (&lt;b&gt;F&lt;/b&gt;) Bar graphs show the measured intensity of GFAP staining from confocal images using ImageJ analysis and (&lt;b&gt;G&lt;/b&gt;) the relative log2-fold change (FC) in expression of the Müller cell gliosis related gene GFAP after 4-OHT treatment for 24 h, 3 days, or 7 days as compared to untreated control organoids. &lt;span class=&quot;html-italic&quot;&gt;n&lt;/span&gt; = 4; * &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.05. Student’s &lt;span class=&quot;html-italic&quot;&gt;t&lt;/span&gt; test; one-way ANOVA.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/10'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g004-550.jpg?1720751470" title=" <strong>Figure 4</strong><br/> &lt;p&gt;Examination of HB-EGF-treated organoids after 4-OHT-induced damage. Representative confocal images of retinal organoids treated with or without 4-OHT for 24 h followed by 7 days treatment with or without HB-EGF. (&lt;b&gt;A&lt;/b&gt;) Sections were stained for cell death and photoreceptor marker expression using recoverin (red) and TUNEL (green) staining. Cell nuclei were detected with DAPI (blue). 40× magnification; scale 50 µm. ONL = outer nuclear layer; INL = inner nuclear layer. (&lt;b&gt;B&lt;/b&gt;) Dot plots show the % of TUNEL-positive cells and proportion of recoverin-positive cells per mm&lt;sup&gt;2&lt;/sup&gt; in treated and untreated organoids. (&lt;b&gt;C&lt;/b&gt;) Box plot shows the average thickness (µm) of retinal layers in organoids 7 days post treatment. &lt;span class=&quot;html-italic&quot;&gt;n&lt;/span&gt; = 3. * &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.05. Student’s &lt;span class=&quot;html-italic&quot;&gt;t&lt;/span&gt; test. One-way ANOVA.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/10'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g005-550.jpg?1720751473" title=" <strong>Figure 5</strong><br/> &lt;p&gt;Examination of Müller glia by immunofluorescence in organoids treated with 4-OHT followed by HB-EGF. Representative confocal images of retinal organoids treated with or without 4-OHT for 24 h followed by 7 days treatment with or without HB-EGF. Sections were stained for (&lt;b&gt;A&lt;/b&gt;) vimentin (green), Ki67 (magenta), and VSX2 (red). (&lt;b&gt;C&lt;/b&gt;) Müller cell responses based on positive staining for vimentin (green), Nestin (red), and SOX2 (magenta). Cell nuclei were detected with DAPI (blue). 40× magnification; scale 50 µm. ONL = outer nuclear layer; INL = inner nuclear layer. (&lt;b&gt;B&lt;/b&gt;) Dot plots show the number of Ki67, VSX2, or double positive cells per mm&lt;sup&gt;2&lt;/sup&gt; in retinal organoids with or without 4-OHT (20 µM) and/or HB-EGF treatment as compared to those without treatment (control) after 7 days. (&lt;b&gt;D&lt;/b&gt;) Dot plot shows the number of SOX2-postive cells per mm&lt;sup&gt;2&lt;/sup&gt; after treatments, and the intensity of nestin or vimentin fluorescence staining was obtained from confocal images using ImageJ analysis. * &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.05, ** &lt;span class=&quot;html-italic&quot;&gt;p&lt;/span&gt; &amp;lt; 0.01; &lt;span class=&quot;html-italic&quot;&gt;n&lt;/span&gt; = 3. Student’s &lt;span class=&quot;html-italic&quot;&gt;t&lt;/span&gt; test; One-way ANOVA.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/10'>Full article</a></strong> "></a><a href="https://pub.mdpi-res.com/organoids/organoids-03-00010/article_deploy/html/images/organoids-03-00010-g006-550.jpg?1720751474" title=" <strong>Figure 6</strong><br/> &lt;p&gt;Gene expression analysis of organoids treated with 4-OHT followed by HB-EGF. Graphs show the relative log2-fold change (FC) in expression of the HB-EGF pathway-related genes PAX6 and NOTCH1 in organoids treated with HB-EGF alone, 4-OHT alone, or 4-OHT and HB-EGF after 7 days as compared to untreated control organoids.&lt;/p&gt; <strong style='display: block; margin-top: 10px; font-size: 18px;'><a style='color: #fff' href='/2674-1172/3/3/10'>Full article</a></strong> "></a></div> </div> </div> </div> <div class="row footer"> <div class="listing-select-options"> <div class="columns small-12"> <div class="select generic-item"> <a href="#" class="export-options-show export-element export-expanded"> Show export options <i class="material-icons">expand_more</i> </a> <a href="#" class="export-options-show export-element"> Show export options <i class="material-icons">expand_less</i> </a> </div> <div class="listing-export-options export-element"> <div class="export-element" style="margin-top: 10px; margin-bottom: 10px;"> <input type="checkbox" class="selector 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