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\u7d30\u80de\u89e3\u96e2\u8a66\u85ac\u306e\u9078\u629e\u30ac\u30a4\u30c9\r\n\r\n\r\n\u8a66\u85ac\r\n\u6982\u8981\r\n\u5099\u8003\r\n\r\n\r\nTrypsin\r\n\u4ee3\u8868\u7684\u306a\u30bf\u30f3\u30d1\u30af\u8cea\u5206\u89e3\u9175\u7d20\r\n\u4e3b\u306b\u30d6\u30bf\u3084\u30a6\u30b7\u306e\u81b5\u81d3\u304b\u3089\u62bd\u51fa\u3055\u308c\u308b\u3002\r\n\u4e0d\u7d14\u7269\u3068\u3057\u3066Trypsin\u4ee5\u5916\u306e\u9175\u7d20\u3082\u542b\u307e\u308c\u308b\u3002\r\n\r\n\r\nTrypsin-EDTA\r\nTrypsin\u3068EDTA\u306e\u6df7\u5408\u7269\r\n\u57f9\u990a\u7d30\u80de\u306e\u7d99\u4ee3\u3067\u6c4e\u7528\u3055\u308c\u3066\u3044\u308b\u3002\r\n\u7d30\u80de\u306e\u7a2e\u985e\u306b\u3088\u3063\u3066\u306f\u3001\u81d3\u5668\u30fb\u7d44\u7e54\u304b\u3089\u306e\r\n\u7d30\u80de\u5358\u96e2\u306b\u9069\u3055\u306a\u3044\u5834\u5408\u304c\u3042\u308b\u3002\r\n\r\n\r\nTrypLE\r\nTrypsin\u69d8\u7d44\u307f\u63db\u3048\u9175\u7d20\u3068EDTA\u306e\u6df7\u5408\u7269\r\nTrypsin-EDTA\u306e\u4ee3\u66ff\u3068\u3057\u3066\u4f7f\u7528\u3067\u304d\u308b\u3002\r\n\u9ad8\u7d14\u5ea6\u30fb\u9ad8\u5b89\u5b9a\u6027\u3067\u5ba4\u6e29\u4fdd\u7ba1\u3067\u304d\u308b\u3002\r\nAnimal Origin Free\u88fd\u54c1\r\n\r\n\r\nCollagenase\r\n\u7d30\u83cc\u7531\u6765\u306e\u30b3\u30e9\u30fc\u30b2\u30f3\u5206\u89e3\u9175\u7d20\r\n\u8907\u6570\u306e\u30d7\u30ed\u30c6\u30a2\u30fc\u30bc\u304c\u542b\u307e\u308c\u3066\u3044\u308b\u3002\r\n\u9175\u7d20\u6d3b\u6027\u306e\u30d0\u30e9\u30f3\u30b9\u306b\u3088\u3063\u3066Type\u5206\u985e\u3055\u308c\u308b\u3002\r\n\r\n\r\nDispase\r\n\u7d30\u83cc\u7531\u6765\u306e\u30bf\u30f3\u30d1\u30af\u8cea\u5206\u89e3\u9175\u7d20\r\n\u57fa\u5e95\u819c\u30bf\u30f3\u30d1\u30af\u8cea\u306e\u5206\u89e3\u306b\u512a\u308c\u3066\u3044\u308b\u3002\r\n\u4f5c\u7528\u304c\u7a4f\u3084\u304b\u3067\u9577\u6642\u9593\u51e6\u7406\u3059\u308b\u3053\u3068\u3082\u3067\u304d\u308b\u3002\r\nAnimal Origin Free\u88fd\u54c1\r\n\r\n\r\nEnzyme-free\r\nEDTA\u3092\u542b\u6709\u3059\u308b\u751f\u7406\u7684\u5869\u985e\u6eb6\u6db2\r\n\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u5207\u65ad\u3057\u306a\u3044\u305f\u3081\u89e3\u96e2\u4f5c\u7528\u306f\u5f31\u3044\u3002\r\nAnimal Origin Free\u88fd\u54c1\r\n\r\n\r\n\r\n\u25bc\u3053\u3093\u306a\u65b9\u306b\u304a\u3059\u3059\u3081\u3067\u3059\uff01\r\n\r\n \t\u30b7\u30f3\u30b0\u30eb\u30bb\u30eb\u89e3\u6790\u306e\u7d30\u80de\u5358\u96e2\u3092\u884c\u3046\u65b9\r\n \t\u521d\u4ee3\u57f9\u990a\u5b9f\u9a13\u306e\u7d30\u80de\u5358\u96e2\u3092\u884c\u3046\u65b9\r\n \t\u7d30\u80de\u57f9\u990a\u3092\u884c\u3046\u65b9\r\n\r\nTrypsin\r\nTrypsin\u306f100\u5e74\u4ee5\u4e0a\u524d\u304b\u3089\u7d30\u80de\u89e3\u96e2\u306b\u5229\u7528\u3055\u308c\u3066\u3044\u308b\u30bf\u30f3\u30d1\u30af\u8cea\u5206\u89e3\u9175\u7d20\u3067\u3059\uff08\u95a2\u9023\u8a18\u4e8b\u300c\u7d30\u80de\u306e\u57f9\u990a\u3068\u7d99\u4ee3\u306e\u30eb\u30fc\u30c4\u3092\u63a2\u3063\u3066\u307f\u305f\u4ef6\uff08\u524d\u7de8\uff09\u300d\uff09\u3002\u30a2\u30eb\u30ae\u30cb\u30f3\u3068\u30ea\u30b7\u30f3\u306e\u30ab\u30eb\u30dc\u30cb\u30eb\u5074\u306e\u30da\u30d7\u30c1\u30c9\u7d50\u5408\u3092\u9078\u629e\u7684\u306b\u5207\u65ad\u3059\u308b\u30a8\u30f3\u30c9\u30da\u30d7\u30c1\u30c0\u30fc\u30bc\u3067\u3001\u7d30\u80de\u9593\u3084\u7d30\u80de\u3068\u57fa\u8cea\u9593\u306e\u7d50\u5408\u3092\u62c5\u3046\u7d30\u80de\u8868\u9762\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u5207\u65ad\u3059\u308b\u3053\u3068\u306b\u3088\u3063\u3066\u7d30\u80de\u3092\u89e3\u96e2\u3055\u305b\u307e\u3059\u3002\u5f53\u793e\u306eGibco\u2122 Trypsin\u88fd\u54c1\u306b\u306f\u3001\u4efb\u610f\u306e\u6fc3\u5ea6\u306b\u6eb6\u89e3\u3057\u3066\u4f7f\u7528\u3059\u308b\u7c89\u672b\u30bf\u30a4\u30d7\u30680.25%\u307e\u305f\u306f2.5%\u306b\u8abf\u88fd\u3055\u308c\u305f\u6eb6\u6db2\u30bf\u30a4\u30d7\u304c\u3042\u308a\u307e\u3059\u30020.25%\u6eb6\u6db2\u306e\u6eb6\u5a92\u306fCa2+\/Mg2+\u30d5\u30ea\u30fc\u306eHBSS\u3067\u3001pH\u6307\u793a\u85ac\u306ephenol red\u304c\u542b\u307e\u308c\u3066\u3044\u307e\u3059\u304c\u30012.5%\u6eb6\u6db2\u306e\u6eb6\u5a92\u306f0.85%\u306eNaCl\u6c34\u6eb6\u6db2\uff08\u751f\u7406\u98df\u5869\u6c34\uff09\u3067\u3059\u3002\u3088\u304f\u77e5\u3089\u308c\u3066\u3044\u308b\u3088\u3046\u306b\u30bf\u30f3\u30d1\u30af\u8cea\u5206\u89e3\u9175\u7d20\u3067\u3042\u308bTrypsin\u81ea\u4f53\u304c\u30bf\u30f3\u30d1\u30af\u8cea\u306e\u305f\u3081\u3001\u4fdd\u7ba1\u72b6\u6cc1\u306b\u3088\u3063\u3066\u306f\u81ea\u5df1\u6d88\u5316\u3057\u3066\u5f90\u3005\u306b\u6d3b\u6027\u304c\u4f4e\u4e0b\u3057\u307e\u3059\u3002\u5b9f\u9a13\u6761\u4ef6\u304c\u56fa\u307e\u308a\u6b21\u7b2c\u30011\u56de\u4f7f\u7528\u5206\u306e\u6db2\u91cf\u3067\u5c0f\u5206\u3051\u3057\u3066\u51b7\u51cd\u3057\u3001\u51cd\u7d50\u878d\u89e3\u3092\u7e70\u308a\u8fd4\u3055\u306a\u3044\u3088\u3046\u306b\u7ba1\u7406\u3059\u308b\u3053\u3068\u304c\u63a8\u5968\u3055\u308c\u307e\u3059\u3002\u3053\u308c\u306fTrypsin\u306b\u9650\u3089\u305a\u3001\u51b7\u51cd\u4fdd\u5b58\u304c\u53ef\u80fd\u306a\u8a66\u85ac\u5168\u822c\u306b\u9069\u7528\u3067\u304d\u308b\u7ba1\u7406\u65b9\u6cd5\u3067\u3059\u3002\u306a\u304a\u3001\u5f53\u793e\u88fd\u54c1\u306eTrypsin\u306f\u30d6\u30bf\u81b5\u81d3\u7531\u6765\u3067\u3059\u3002\u88682\u306b\u5f53\u793e\u306eTrypsin\u88fd\u54c1\u3092\u307e\u3068\u3081\u307e\u3057\u305f\u3002\r\n\u88682. Trypsin\u88fd\u54c1\u306e\u9078\u629e\u30ac\u30a4\u30c9\r\n\r\n\r\n\u88fd\u54c1\u756a\u53f7\r\nVolume\r\nTrypsin\r\nPhenol\r\nRed\r\nBuffer\r\n\r\n\r\n27250018\r\n100 g\r\nPowder\r\nX\r\nX\r\n\r\n\r\n15050065\r\n100 mL\r\n0.25\uff05\r\n\u3007\r\nHBSS w\/o Ca2+ & Mg2+\r\n\r\n\r\n15050057\r\n500 mL\r\n\r\n\r\n15090046\r\n100 mL\r\n2.5\uff05\r\nX\r\n0.85% NaCl\r\n\r\n\r\n\r\nTrypsin-EDTA\r\nTrypsin-EDTA\u306f\u30bf\u30f3\u30d1\u30af\u8cea\u5206\u89e3\u9175\u7d20\u306eTrypsin\u3068Ca2+\u3084Mg2+\u306a\u3069\u306e2\u4fa1\u306e\u967d\u30a4\u30aa\u30f3\u3092\u30ad\u30ec\u30fc\u30c8\u3059\u308b\u30a8\u30c1\u30ec\u30f3\u30b8\u30a2\u30df\u30f3\u56db\u9162\u9178\uff08ethylenediaminetetraacetic acid\uff09\u306e\u6df7\u5408\u6eb6\u6db2\u3067\u3059\u3002Ca2+\u3068Mg2+\u306f\u7d30\u80de\u9593\u3084\u7d30\u80de-\u57fa\u8cea\u9593\u306e\u63a5\u7740\u306b\u5fc5\u8981\u3067\u3042\u308b\u3053\u3068\u304b\u3089\u3001\u3053\u308c\u3089\u306e\u30a4\u30aa\u30f3\u3092\u30ad\u30ec\u30fc\u30c8\u3059\u308b\u3053\u3068\u306b\u3088\u3063\u3066\u7d30\u80de\u89e3\u96e2\u3092\u4fc3\u9032\u3057\u307e\u3059\u3002Trypsin-EDTA\u306f1950\u5e74\u4ee3\u306b\u57f9\u990a\u7d30\u80de\u306e\u89e3\u96e2\u306b\u6709\u7528\u3067\u3042\u308b\u3053\u3068\u304c\u5831\u544a\u3055\u308c\u307e\u3057\u305f\uff08\u95a2\u9023\u8a18\u4e8b\u300c\u7d30\u80de\u306e\u57f9\u990a\u3068\u7d99\u4ee3\u306e\u30eb\u30fc\u30c4\u3092\u63a2\u3063\u3066\u307f\u305f\u4ef6\uff08\u5f8c\u7de8\uff09\u300d\uff09\u3002\u305d\u308c\u4ee5\u6765\u3001\u4e3b\u306b\u63a5\u7740\u57f9\u990a\u7d30\u80de\u306e\u7d99\u4ee3\u6642\u306b\u4f7f\u7528\u3055\u308c\u308b\u8a66\u85ac\u3067\u3059\u304c\u3001\u81d3\u5668\u30fb\u7d44\u7e54\u306e\u7d30\u80de\u5206\u6563\u306b\u4f7f\u7528\u3055\u308c\u308b\u3053\u3068\u3082\u3042\u308a\u307e\u3059\u3002\u305f\u3060\u3001\u7d30\u80de\u306e\u7a2e\u985e\u306b\u3088\u3063\u3066\u306fEDTA\u304c\u7d30\u80de\u306b\u30c0\u30e1\u30fc\u30b8\u3092\u4e0e\u3048\u308b\u30b1\u30fc\u30b9\u304c\u3042\u308b\u3088\u3046\u3067\u3059\u3002EDTA\u4e0d\u542b\u306eTrypsin\u6eb6\u6db2\u3067\u78ba\u7acb\u3055\u308c\u305f\u7d30\u80de\u89e3\u96e2\u64cd\u4f5c\u3067\u306fTrypsin-EDTA\u3067\u7f6e\u304d\u63db\u3048\u3067\u304d\u306a\u3044\u3053\u3068\u304c\u3042\u308b\u305f\u3081\u6ce8\u610f\u304c\u5fc5\u8981\u3067\u3059\u3002\u3053\u306e\u3053\u3068\u306f\u3001\u5f8c\u8ff0\u3059\u308bTrypLE\u306b\u3082\u5f53\u3066\u306f\u307e\u308a\u307e\u3059\u3002\u5f53\u793e\u306eGibco\u2122 Trypsin-EDTA\u88fd\u54c1\u3092\u88683\u306b\u307e\u3068\u3081\u307e\u3057\u305f\u3002\r\n\u88683. Trypsin-EDTA\u88fd\u54c1\u306e\u9078\u629e\u30ac\u30a4\u30c9\r\n\r\n\r\n\u88fd\u54c1\u756a\u53f7\r\nVolume\r\nTrypsin\r\nEDTA\r\nPhenol\r\nRed\r\nBuffer\r\n\r\n\r\n25300054\r\n100 mL\r\n0.05\uff05\r\n0.48 mM\r\n\u25cb\r\nHBSS w\/o \u2026","hashtag":"ThermoFisherJP","subscribe":"content","subscribe_url":"","activestatus":"1","singular":"1","twitter_popup":"1","refresh":"0","nonce":"ac99902330","postid":"16245","servertime":"1732982251","ajaxurl":"https:\/\/www.thermofisher.com\/blog\/wp-admin\/admin-ajax.php"}; /* ]]> */ </script> <script type="text/javascript" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/plugins/mashsharer/assets/js/mashsb.min.js?ver=4.0.47" id="mashsb-js"></script> <script type="text/javascript" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/plugins/mashshare-google-analytics/assets/js/mashga.min.js?ver=1.0.9" id="mashga-js"></script> <script type="text/javascript" id="mashnet-js-extra"> /* <![CDATA[ */ var mashnet = {"body":"Check out this article I just read:","subject":"Check out this article from Accelerating Science","pinterest_select":"1","pinterest_image":"https:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/wp-content\/uploads\/sites\/13\/2024\/01\/cell-culture-cell-separation-bid-ts-mbr-24001-image.png","pinterest_desc":"\u8fd1\u5e74\u306e\u6025\u6fc0\u306a\u6280\u8853\u9032\u6b69\u306b\u3088\u308b\u6b21\u4e16\u4ee3\u30b7\u30fc\u30b1\u30f3\u31f2\u306e\u30c7\u30fc\u30bf\u53d6\u5f97\u30b3\u30b9\u30c8\u306e\u4f4e\u4e0b\u306b\u3088\u308a\u3001\u30b7\u30f3\u30b0\u30eb\u30bb\u30ebRNA\u30b7\u30fc\u30b1\u30f3\u30b7\u30f3\u30b0\uff08single-cell RNA-seq\u3001scRNA-seq\uff09\u306b\u4ee3\u8868\u3055\u308c\u308b\u30b7\u30f3\u30b0\u30eb\u30bb\u30eb\u89e3\u6790\u306f\u307e\u3059\u307e\u3059\u8eab\u8fd1\u306a\u5b9f\u9a13\u624b\u6cd5\u3068\u306a\u308a\u3064\u3064\u3042\u308a\u307e\u3059\u3002Google Scholar\u306b\u3088\u308b\u3068\u3001\u201dsingle-cell 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\u7d30\u80de\u89e3\u96e2\u8a66\u85ac\u306e\u9078\u629e\u30ac\u30a4\u30c9\r\n\r\n\r\n\u8a66\u85ac\r\n\u6982\u8981\r\n\u5099\u8003\r\n\r\n\r\nTrypsin\r\n\u4ee3\u8868\u7684\u306a\u30bf\u30f3\u30d1\u30af\u8cea\u5206\u89e3\u9175\u7d20\r\n\u4e3b\u306b\u30d6\u30bf\u3084\u30a6\u30b7\u306e\u81b5\u81d3\u304b\u3089\u62bd\u51fa\u3055\u308c\u308b\u3002\r\n\u4e0d\u7d14\u7269\u3068\u3057\u3066Trypsin\u4ee5\u5916\u306e\u9175\u7d20\u3082\u542b\u307e\u308c\u308b\u3002\r\n\r\n\r\nTrypsin-EDTA\r\nTrypsin\u3068EDTA\u306e\u6df7\u5408\u7269\r\n\u57f9\u990a\u7d30\u80de\u306e\u7d99\u4ee3\u3067\u6c4e\u7528\u3055\u308c\u3066\u3044\u308b\u3002\r\n\u7d30\u80de\u306e\u7a2e\u985e\u306b\u3088\u3063\u3066\u306f\u3001\u81d3\u5668\u30fb\u7d44\u7e54\u304b\u3089\u306e\r\n\u7d30\u80de\u5358\u96e2\u306b\u9069\u3055\u306a\u3044\u5834\u5408\u304c\u3042\u308b\u3002\r\n\r\n\r\nTrypLE\r\nTrypsin\u69d8\u7d44\u307f\u63db\u3048\u9175\u7d20\u3068EDTA\u306e\u6df7\u5408\u7269\r\nTrypsin-EDTA\u306e\u4ee3\u66ff\u3068\u3057\u3066\u4f7f\u7528\u3067\u304d\u308b\u3002\r\n\u9ad8\u7d14\u5ea6\u30fb\u9ad8\u5b89\u5b9a\u6027\u3067\u5ba4\u6e29\u4fdd\u7ba1\u3067\u304d\u308b\u3002\r\nAnimal 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href="https://www.thermofisher.com/blog/learning-at-the-bench/contact-us/"><span >お問い合わせ</span></a></li> </ul></nav></div></section> </div><div class="breadcrumb"><span class="breadcrumb-link-wrap"><a href="https://www.thermofisher.com/blog/?">Accelerating Science</a></span><span class="breadcrumb-link-wrap"><a class="breadcrumb-link" href="https://www.thermofisher.com/blog/learning-at-the-bench/"><span class="breadcrumb-link-text-wrap">Learning at the Bench</span></a><meta ></span> <span aria-label="breadcrumb separator">/</span> <span class="breadcrumb-link-wrap"><a class="breadcrumb-link" href="https://www.thermofisher.com/blog/learning-at-the-bench/cell/"><span class="breadcrumb-link-text-wrap">細胞培養・イメージング</span></a><meta ></span> <span aria-label="breadcrumb separator">/</span> <span class="breadcrumb-current-title">シングルセル解析や初代培養に使われる細胞解離試薬の選択ガイド</span></div></div></header><div class="site-inner"><div class="content-sidebar-wrap"><main class="content"><article class="post-16245 post type-post status-publish format-standard has-post-thumbnail category-cell tag-cell-separation tag-cell-culture entry" aria-label="シングルセル解析や初代培養に使われる細胞解離試薬の選択ガイド"><header class="entry-header"><h1 class="entry-title">シングルセル解析や初代培養に使われる細胞解離試薬の選択ガイド</h1> <span class="post-info-author"> 作成者 <a href="https://www.thermofisher.com/blog/author/lgbadmin/" title="LatB Staff の投稿" rel="author">LatB Staff</a> </span> <span class="post-info-date"> 01.10.2024 </span> </header><div class="entry-content"><p>近年の急激な技術進歩による次世代シーケンㇲのデータ取得コストの低下により、シングルセルRNAシーケンシング（single-cell RNA-seq、scRNA-seq）に代表されるシングルセル解析はますます身近な実験手法となりつつあります。Google Scholarによると、”single-cell RNA-seq”のキーワードでヒットする約3万件の文献のうち2万件は2022年以降に発表されました。同じく2022年以降に発表された文献の検索で、”real-time PCR”が3万7000件、”genome editing”が2万3000件、”western blot”が2万2000件のヒットであることから、scRNA-seqの普及がこれらの代表的な実験手法に迫る勢いであることが伺えます（2023年11月時点）。</p> <p>浮遊性の細胞や接着性の培養細胞を解析対象とする場合は、scRNA-seqのための単離細胞の取得は比較的容易です。一方、生体の臓器や組織を解析対象とする場合は、酵素処理などによって細胞を単離する必要があります。細胞へのストレスを最小限に抑えながら臓器・組織を分散させる実験条件は、対象の動物種、個体の成熟度（生育期間や発生段階）、細胞の種類などによって最適化する必要があります。そのため、幅広い細胞種に対応できる細胞単離キットの開発や機械による自動化は容易ではないと考えられ、現在でも多くの研究者がさまざまな試薬を駆使して手作業で単離細胞を調製しています。</p> <p>そこで当記事では、RNA-seqのための単離細胞調製への実績のある製品を中心に、当社の細胞解離試薬をまとめました。当記事で扱っている試薬の概要を表1に整理しました。各試薬ごとにご紹介していますので、特定の試薬にご興味のある方は、当記事の該当部分へお進みください。</p> <table width="633"> <caption>表1. 細胞解離試薬の選択ガイド</caption> <tbody> <tr bgcolor="#696969"> <td style="text-align: center" width="137"><span style="color: #ffffff">試薬</span></td> <td style="text-align: center" width="220"><span style="color: #ffffff">概要</span></td> <td style="text-align: center" width="303"><span style="color: #ffffff">備考</span></td> </tr> <tr> <td width="137"><a href="#Trypsin">Trypsin</a></td> <td width="220">代表的なタンパク質分解酵素</td> <td width="303">主にブタやウシの膵臓から抽出される。<br /> 不純物としてTrypsin以外の酵素も含まれる。</td> </tr> <tr> <td width="137"><a href="#Trypsin-EDTA">Trypsin-EDTA</a></td> <td width="220">TrypsinとEDTAの混合物</td> <td width="303">培養細胞の継代で汎用されている。<br /> 細胞の種類によっては、臓器・組織からの<br /> 細胞単離に適さない場合がある。</td> </tr> <tr> <td width="137"><a href="#TrypLE">TrypLE</a></td> <td width="220">Trypsin様組み換え酵素とEDTAの混合物</td> <td width="303">Trypsin-EDTAの代替として使用できる。<br /> 高純度・高安定性で室温保管できる。<br /> Animal Origin Free製品</td> </tr> <tr> <td width="137"><a href="#Collagenase">Collagenase</a></td> <td width="220">細菌由来のコラーゲン分解酵素</td> <td width="303">複数のプロテアーゼが含まれている。<br /> 酵素活性のバランスによってType分類される。</td> </tr> <tr> <td width="137"><a href="#Dispase">Dispase</a></td> <td width="220">細菌由来のタンパク質分解酵素</td> <td width="303">基底膜タンパク質の分解に優れている。<br /> 作用が穏やかで長時間処理することもできる。<br /> Animal Origin Free製品</td> </tr> <tr> <td width="137"><a href="#Enzyme-free">Enzyme-free</a></td> <td width="220">EDTAを含有する生理的塩類溶液</td> <td width="303">タンパク質を切断しないため解離作用は弱い。<br /> Animal Origin Free製品</td> </tr> </tbody> </table> <p>▼こんな方におすすめです！</p> <ul> <li>シングルセル解析の細胞単離を行う方</li> <li>初代培養実験の細胞単離を行う方</li> <li>細胞培養を行う方</li> </ul> <div id="toc_container" class="no_bullets"><p class="toc_title">▼もくじ</p><ul class="toc_list"><li><a href="#Trypsin">Trypsin</a></li><li><a href="#Trypsin-EDTA">Trypsin-EDTA</a></li><li><a href="#TrypLE">TrypLEシリーズ</a></li><li><a href="#Collagenase">Collagenase</a></li><li><a href="#Dispase">Dispase</a></li><li><a href="#i">酵素フリーの細胞解離試薬</a></li><li><a href="#Certificate_of_AnalysisCOA">（補足）分析証明書（Certificate of Analysis、COA）の入手方法</a></li><li><a href="#i-2">まとめ</a><ul><li><a href="#3D">【無料ダウンロード】3D細胞培養＆解析ハンドブック</a></li><li><a href="#Gibco">【無料ダウンロード】Gibco多能性幹細胞ハンドブック</a></li></ul></li></ul></div> <h2><span id="Trypsin">Trypsin</span></h2> <p>Trypsinは100年以上前から細胞解離に利用されているタンパク質分解酵素です（関連記事「<a href="https://www.thermofisher.com/blog/learning-at-the-bench/cell_culture_history_1_bid_ts_1/?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener">細胞の培養と継代のルーツを探ってみた件（前編）</a>」）。アルギニンとリシンのカルボニル側のペプチド結合を選択的に切断するエンドペプチダーゼで、細胞間や細胞と基質間の結合を担う細胞表面タンパク質を切断することによって細胞を解離させます。当社のGibco™ Trypsin製品には、任意の濃度に溶解して使用する粉末タイプと0.25%または2.5%に調製された溶液タイプがあります。0.25%溶液の溶媒はCa<sup>2+</sup>/Mg<sup>2+</sup>フリーのHBSSで、pH指示薬のphenol redが含まれていますが、2.5%溶液の溶媒は0.85%のNaCl水溶液（生理食塩水）です。よく知られているようにタンパク質分解酵素であるTrypsin自体がタンパク質のため、保管状況によっては自己消化して徐々に活性が低下します。実験条件が固まり次第、1回使用分の液量で小分けして冷凍し、凍結融解を繰り返さないように管理することが推奨されます。これはTrypsinに限らず、冷凍保存が可能な試薬全般に適用できる管理方法です。なお、当社製品のTrypsinはブタ膵臓由来です。表2に当社のTrypsin製品をまとめました。</p> <table width="537"> <caption>表2. Trypsin製品の選択ガイド</caption> <tbody> <tr bgcolor="#696969"> <td style="text-align: center" width="81"><span style="color: #ffffff">製品番号</span></td> <td style="text-align: center" width="96"><span style="color: #ffffff">Volume</span></td> <td style="text-align: center" width="87"><span style="color: #ffffff">Trypsin</span></td> <td style="text-align: center" width="83"><span style="color: #ffffff">Phenol</span><br /> <span style="color: #ffffff">Red</span></td> <td style="text-align: center" width="189"><span style="color: #ffffff">Buffer</span></td> </tr> <tr> <td style="text-align: center" width="81">27250018</td> <td style="text-align: center" width="96">100 g</td> <td style="text-align: center" width="87">Powder</td> <td style="text-align: center" width="83">X</td> <td style="text-align: center" width="189">X</td> </tr> <tr> <td style="text-align: center" width="81">15050065</td> <td style="text-align: center" width="96">100 mL</td> <td style="text-align: center" rowspan="2" width="87">0.25％</td> <td style="text-align: center" rowspan="2" width="83">〇</td> <td style="text-align: center" rowspan="2" width="189">HBSS w/o Ca<sup>2+</sup> & Mg<sup>2+</sup></td> </tr> <tr> <td style="text-align: center" width="81">15050057</td> <td style="text-align: center" width="96">500 mL</td> </tr> <tr> <td style="text-align: center" width="81">15090046</td> <td style="text-align: center" width="96">100 mL</td> <td style="text-align: center" width="87">2.5％</td> <td style="text-align: center" width="83">X</td> <td style="text-align: center" width="189">0.85% NaCl</td> </tr> </tbody> </table> <h2><span id="Trypsin-EDTA">Trypsin-EDTA</span></h2> <p>Trypsin-EDTAはタンパク質分解酵素のTrypsinとCa<sup>2+</sup>やMg<sup>2+</sup>などの2価の陽イオンをキレートするエチレンジアミン四酢酸（ethylenediaminetetraacetic acid）の混合溶液です。Ca<sup>2+</sup>とMg<sup>2+</sup>は細胞間や細胞-基質間の接着に必要であることから、これらのイオンをキレートすることによって細胞解離を促進します。Trypsin-EDTAは1950年代に培養細胞の解離に有用であることが報告されました（関連記事「<a href="https://www.thermofisher.com/blog/learning-at-the-bench/cell_culture_history_2_bid_ts_2/?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener">細胞の培養と継代のルーツを探ってみた件（後編）</a>」）。それ以来、主に接着培養細胞の継代時に使用される試薬ですが、臓器・組織の細胞分散に使用されることもあります。ただ、細胞の種類によってはEDTAが細胞にダメージを与えるケースがあるようです。EDTA不含のTrypsin溶液で確立された細胞解離操作ではTrypsin-EDTAで置き換えできないことがあるため注意が必要です。このことは、後述するTrypLEにも当てはまります。当社のGibco™ Trypsin-EDTA製品を表3にまとめました。</p> <table width="633"> <caption>表3. Trypsin-EDTA製品の選択ガイド</caption> <tbody> <tr bgcolor="#696969"> <td style="text-align: center" width="85"><span style="color: #ffffff">製品番号</span></td> <td style="text-align: center" width="123"><span style="color: #ffffff">Volume</span></td> <td style="text-align: center" width="85"><span style="color: #ffffff">Trypsin</span></td> <td style="text-align: center" width="79"><span style="color: #ffffff">EDTA</span></td> <td style="text-align: center" width="83"><span style="color: #ffffff">Phenol</span><br /> <span style="color: #ffffff">Red</span></td> <td style="text-align: center" width="179"><span style="color: #ffffff">Buffer</span></td> </tr> <tr> <td style="text-align: center" width="85">25300054</td> <td style="text-align: center" width="123">100 mL</td> <td style="text-align: center" rowspan="3" width="85">0.05％</td> <td style="text-align: center" rowspan="3" width="79">0.48 mM</td> <td style="text-align: center" rowspan="3" width="83">○</td> <td style="text-align: center" rowspan="3" width="179">HBSS w/o Ca<sup>2+</sup> & Mg<sup>2+</sup></td> </tr> <tr> <td style="text-align: center" width="85">25300062</td> <td style="text-align: center" width="123">500 mL</td> </tr> <tr> <td style="text-align: center" width="85">25300120</td> <td style="text-align: center" width="123">20 x 100 mL</td> </tr> <tr> <td style="text-align: center" width="85">25200056</td> <td style="text-align: center" width="123">100 mL</td> <td style="text-align: center" rowspan="3" width="85">0.25％</td> <td style="text-align: center" rowspan="3" width="79">0.91 mM</td> <td style="text-align: center" rowspan="3" width="83">○</td> <td style="text-align: center" rowspan="3" width="179">HBSS w/o Ca<sup>2+</sup> & Mg<sup>2+</sup></td> </tr> <tr> <td style="text-align: center" width="85">25200072</td> <td style="text-align: center" width="123">500 mL</td> </tr> <tr> <td style="text-align: center" width="85">25200114</td> <td style="text-align: center" width="123">20 x 100 mL</td> </tr> <tr> <td style="text-align: center" width="85">15400054</td> <td style="text-align: center" width="123">100 mL</td> <td style="text-align: center" width="85">0.5％</td> <td style="text-align: center" width="79">4.8 mM</td> <td style="text-align: center" width="83">X</td> <td style="text-align: center" width="179">0.85% NaCl</td> </tr> <tr> <td style="text-align: center" width="85">R001100</td> <td style="text-align: center" width="123">100 mL</td> <td style="text-align: center" width="85">0.025％</td> <td style="text-align: center" width="79">0.3 mM</td> <td style="text-align: center" width="83">○</td> <td style="text-align: center" width="179">※</td> </tr> </tbody> <tfoot> <tr> <td style="border-bottom: none" colspan="6" width="633">※組成は<a href="https://www.thermofisher.com/jp/ja/home/technical-resources/media-formulation.338.html?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener">こちら</a>をご確認ください。</td> </tr> </tfoot> </table> <h2><span id="TrypLE">TrypLEシリーズ</span></h2> <p>Gibco™ <a href="https://www.thermofisher.com/jp/ja/home/life-science/cell-culture/mammalian-cell-culture/reagents/trypsin/tryple-express.html?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener">TrypLE™シリーズ</a>はTrypsin-EDTA溶液の代替品として使用できる細胞解離酵素試薬です。微生物に産生させたTrypsin様の組換え酵素を高純度で精製して製造しているため、膵臓由来の不純物の酵素が残存しているTrypsinと比べて細胞にやさしいとされています。動物由来成分を含まない製品（Animal Origin Free、AOF）であり、TrypLE溶液と同量以上の、無血清培地（血清入り培地でも可）や生理的塩類溶液で希釈することで反応を止めることができるため、無血清培地を用いた細胞培養アプリケーションに使用できます。また、TrypLEは常温で保管できるため取扱いの面でも便利です（冷蔵・冷凍保管も可）。</p> <p>TrypLEシリーズには酵素濃度の違い（1Xまたは10X）やPhenol Redの有無に加えて「Express」「Select」「CTS Select」の3種類の分類があります。これらの3種類に含まれるTrypLEの酵素としての性能は同等ですが、目的によって使い分けがあります。</p> <ul> <li>Gibco™ TrypLE™ Express Enzyme：研究用（Research Use Only、RUO）の標準グレード</li> <li>Gibco™ TrypLE™ Select Enzyme：研究用・工業原料用のグレード</li> <li>Gibco™ CTS™ TrypLE Select Enzyme：再生医療研究・臨床研究などのためのグレード</li> </ul> <p>一般的な研究用途の場合はTrypLE Express Enzymeをお使いください。製造などを視野に入れている場合は、最先端の施設において専用の装置で製造されているTrypLE Select Enzymeが推奨します。CTS TrypLE Select Enzymeの「CTS」はCell Therapy Systemsの略語で、基礎研究から臨床研究までをサポートするための製品シリーズです。CTSシリーズの特徴や製品については<a href="https://assets.thermofisher.com/TFS-Assets/BID/brochures/gibco-cts-brochure-ja.pdf?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener">こちら</a>の資料をご参照ください。なおCTSシリーズの製品は「再生医療等製品材料適格性確認書」が取得されています（該当製品一覧と問い合わせフォームは<a href="https://www.thermofisher.com/jp/ja/home/global/forms/sbi-cert-inquiry.html?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener">こちら</a>）。表4でTrypLE製品を整理しています。</p> <table width="633"> <caption>表4. TrypLE製品の選択ガイド</caption> <tbody> <tr bgcolor="#696969"> <td style="text-align: center" width="85"><span style="color: #ffffff">製品番号</span></td> <td style="text-align: center" width="102"><span style="color: #ffffff">Volume</span></td> <td style="text-align: center" width="138"><span style="color: #ffffff">製品名</span></td> <td style="text-align: center" width="113"><span style="color: #ffffff">TrypLE<br /> Concentration</span></td> <td style="text-align: center" width="72"><span style="color: #ffffff">EDTA</span></td> <td style="text-align: center" width="74"><span style="color: #ffffff">Phenol<br /> Red</span></td> <td style="text-align: center" width="49"><span style="color: #ffffff">Buffer</span></td> </tr> <tr> <td style="text-align: center" width="85">12604013</td> <td style="text-align: center" width="102">100 mL</td> <td style="text-align: center" rowspan="4" width="138">TrypLE Express Enzyme</td> <td style="text-align: center" rowspan="4" width="113">1X</td> <td style="text-align: center" rowspan="4" width="72">1.1 mM</td> <td style="text-align: center" rowspan="4" width="74">X</td> <td style="text-align: center" rowspan="4" width="49">DPBS</td> </tr> <tr> <td style="text-align: center" width="85">12604021</td> <td style="text-align: center" width="102">500 mL</td> </tr> <tr> <td style="text-align: center" width="85">12604039</td> <td style="text-align: center" width="102">20 x 100 mL</td> </tr> <tr> <td style="text-align: center" width="85">12604054</td> <td style="text-align: center" width="102">5000 mL</td> </tr> <tr> <td style="text-align: center" width="85">12605010</td> <td style="text-align: center" width="102">100 mL</td> <td style="text-align: center" rowspan="4" width="138">TrypLE Express Enzyme</td> <td style="text-align: center" rowspan="4" width="113">1X</td> <td style="text-align: center" rowspan="4" width="72">1.1 mM</td> <td style="text-align: center" rowspan="4" width="74">○</td> <td style="text-align: center" rowspan="4" width="49">DPBS</td> </tr> <tr> <td style="text-align: center" width="85">12605028</td> <td style="text-align: center" width="102">500 mL</td> </tr> <tr> <td style="text-align: center" width="85">12605036</td> <td style="text-align: center" width="102">20 x 100 mL</td> </tr> <tr> <td style="text-align: center" width="85">12605093</td> <td style="text-align: center" width="102">5000</td> </tr> <tr> <td style="text-align: center" width="85">12563011</td> <td style="text-align: center" width="102">100</td> <td style="text-align: center" rowspan="2" width="138">TrypLE Select Enzyme</td> <td style="text-align: center" rowspan="2" width="113">1X</td> <td style="text-align: center" rowspan="2" width="72">1.1 mM</td> <td style="text-align: center" rowspan="2" width="74">X</td> <td style="text-align: center" rowspan="2" width="49">DPBS</td> </tr> <tr> <td style="text-align: center" width="85">12563029</td> <td style="text-align: center" width="102">500</td> </tr> <tr> <td style="text-align: center" width="85">A1217701</td> <td style="text-align: center" width="102">100</td> <td style="text-align: center" rowspan="3" width="138">TrypLE Select Enzyme</td> <td style="text-align: center" rowspan="3" width="113"><span style="color: #ff0000">10X</span></td> <td style="text-align: center" rowspan="3" width="72">1.1 mM</td> <td style="text-align: center" rowspan="3" width="74">X</td> <td style="text-align: center" rowspan="3" width="49">DPBS</td> </tr> <tr> <td style="text-align: center" width="85">A1217702</td> <td style="text-align: center" width="102">500</td> </tr> <tr> <td style="text-align: center" width="85">A1217703</td> <td style="text-align: center" width="102">20 x 100 mL</td> </tr> <tr> <td style="text-align: center" width="85">A1285901</td> <td style="text-align: center" width="102">100</td> <td style="text-align: center" rowspan="2" width="138"><span style="color: #ff0000">CTS</span> TrypLE Select Enzyme</td> <td style="text-align: center" rowspan="2" width="113">1X</td> <td style="text-align: center" rowspan="2" width="72">1.1 mM</td> <td style="text-align: center" rowspan="2" width="74">X</td> <td style="text-align: center" rowspan="2" width="49">DPBS</td> </tr> <tr> <td style="text-align: center" width="85">A4738001</td> <td style="text-align: center" width="102">1000</td> </tr> </tbody> </table> <p>“10X”のTrypLE Select Enzymeは接着が強固な細胞の分散にそのまま使用できるように、EDTAの濃度やDPBSの組成は”1X”と同等です。したがって、任意の濃度に希釈して使用する場合は、適切な濃度のEDTAを含むDPBSを用いる必要があります。詳しい手順は<a href="https://tools.thermofisher.com/content/sfs/manuals/TrypLE_man_JP.pdf?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener">製品マニュアル</a>に記載されていますのでご使用の際はご確認ください。</p> <h2><span id="Collagenase">Collagenase</span></h2> <p>多細胞生物の組織はコラーゲン、フィブロネクチン、ラミニンなどの細胞外基質（ECM）を豊富に含んでいます。Collagenaseは組織中に最も多く存在するECMであるコラーゲンを分解するタンパク質分解酵素の総称です。当社が販売しているGibco™ Collagenaseはヒストリチクム菌（Clostridium histolyticum）から分離精製された細菌性コラゲナーゼの粉末製品です。このCollagenaseは、コラーゲン分子に繰り返し出現する「Proline-X（中性アミノ酸）-Glycine-Proline」の配列のXとGlycineの間を切断します。一般に三重らせん構造をもつ天然コラーゲン線維は生理的条件下においてTrypsinなどのタンパク質分解酵素に耐性を示します。そのため、コラーゲンを豊富に含む組織からの細胞の単離にCollagenaseが使用されることがあります。表5に当社のCollagenase製品をまとめました。</p> <table width="783"> <caption>表5. Collagenase製品の選択ガイド</caption> <tbody> <tr bgcolor="#696969"> <td style="text-align: center" width="81"><span style="color: #ffffff">製品番号</span></td> <td style="text-align: center" width="173"><span style="color: #ffffff">製品名</span></td> <td style="text-align: center" width="87"><span style="color: #ffffff">Size</span></td> <td style="text-align: center" width="181"><span style="color: #ffffff">Tissue/Cell type</span></td> <td style="text-align: center" width="261"><span style="color: #ffffff">Memo</span></td> </tr> <tr> <td style="text-align: center" width="81">17100017</td> <td style="text-align: center" width="173">Collagenase, Type I</td> <td style="text-align: center" width="87">1 g</td> <td style="text-align: center" rowspan="2" width="181">上皮、副腎、肺、<br /> 脂肪、肝臓</td> <td style="text-align: center" rowspan="2" width="261">collagenase、caseinase、<br /> clostripain、trypsinの<br /> 活性がバランスよく含まれる</td> </tr> <tr> <td style="text-align: center" width="81">17018029</td> <td style="text-align: center" width="173">Collagenase, Type I</td> <td style="text-align: center" width="87">500 mg</td> </tr> <tr> <td style="text-align: center" width="81">17101015</td> <td style="text-align: center" width="173">Collagenase, Type II</td> <td style="text-align: center" width="87">1 g</td> <td style="text-align: center" width="181">心臓、甲状腺、唾液腺、肝臓、骨、軟骨</td> <td style="text-align: center" width="261">clostripainの活性が比較的高い<br /> clostripainはArg残基のC末側を<br /> 切断するエンドペプチダーゼ</td> </tr> <tr> <td style="text-align: center" width="81">17104019</td> <td style="text-align: center" width="173">Collagenase, Type IV</td> <td style="text-align: center" width="87">1 g</td> <td style="text-align: center" width="181">膵島</td> <td style="text-align: center" width="261">trypsin活性が比較的低い</td> </tr> </tbody> </table> <p>表5のMemo欄にあるように、Collagenase製品にはcollagenase以外にcaseinase、clostripain、trypsinなどの活性が含まれています。これらの活性のバランスによってType I、II、IVに分類されており、それぞれに適した臓器・組織の種類が知られています。表中の記載は代表例ですので、記載以外のさまざまな組織への使用事例があります。Collagenase製品に含まれる複数の酵素活性は、効率的な細胞解離に寄与するとともに細胞へのダメージの原因にもなりえます。重量当たりの酵素活性はロットごとに異なるため、分析証明書に記載されている酵素活性を参考にユニット濃度（U/mL）を設定して溶解するとともに、ロット変更時にはロットチェックを行うことが推奨されます。なお、分析証明書の入手方法については追って触れます。</p> <h2><span id="Dispase">Dispase</span></h2> <p>Gibco™ <a href="https://www.thermofisher.com/order/catalog/product/17105041?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener">Dispase II</a>はグラム陽性細菌の1種であるPaenibacillus polymyxaが産生するアミノエンドペプチダーゼです。非極性アミノ酸残基のN末側のペプチド結合を切断します。酵素活性が比較的穏やかで細胞傷害が少ない、血清入り培地中で酵素活性を発揮する、自身を分解しないため長時間の反応が可能などの有用な特徴があります。基底膜を構成するECMタンパク質をよく分解するため、適切に処理すると上皮組織と間質組織を分離させることができます（動画）。Collagenaseと同様にロットごとに酵素活性が異なるため、分析証明書を確認して粉末を溶解してください。</p> <div style="width: 400px;" class="wp-video"> <video class="wp-video-shortcode" id="video-16245-1" width="400" height="400" poster="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/01/cell-culture-cell-separation-bid-ts-mbr-24001-image.png" loop="1" preload="metadata" controls="controls"><source type="video/mp4" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/01/cell-culture-cell-separation-bid-ts-mbr-24001-01.mp4?_=1" /><a href="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/01/cell-culture-cell-separation-bid-ts-mbr-24001-01.mp4">https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/01/cell-culture-cell-separation-bid-ts-mbr-24001-01.mp4</a></video></div> <h2 id="Enzyme-free"><span id="i">酵素フリーの細胞解離試薬</span></h2> <p>以上は、いずれもタンパク質分解酵素の活性によって細胞接着を切断して細胞を解離させる製品でした。最後に、EDTAによるキレート作用によって細胞を解離させる酵素フリーの製品をご案内します。これまでの製品と比べて細胞解離作用が弱いため、臓器・組織の解離や、接着の強い培養細胞の解離には適していません。逆に、タンパク質分解酵素を含まないため、細胞表面のタンパク質を傷つけずに解析する必要がある場合などに有用です。当社の製品を表6にまとめました。</p> <table width="723"> <caption>表6.酵素フリーの細胞解離試薬の選択ガイド</caption> <tbody> <tr bgcolor="#696969"> <td style="text-align: center" width="89"><span style="color: #ffffff">製品番号</span></td> <td style="text-align: center" width="297"><span style="color: #ffffff">製品名</span></td> <td style="text-align: center" width="87"><span style="color: #ffffff">Volume</span></td> <td style="text-align: center" width="91"><span style="color: #ffffff">EDTA</span></td> <td style="text-align: center" width="159"><span style="color: #ffffff">Buffer</span></td> </tr> <tr> <td style="text-align: center" width="89">15040066</td> <td style="text-align: center" width="297">Gibco™ Versene Solution</td> <td style="text-align: center" width="87">100 mL</td> <td style="text-align: center" width="91">0.48 mM</td> <td style="text-align: center" width="159">PBS</td> </tr> <tr> <td style="text-align: center" width="89">A4239101</td> <td style="text-align: center" width="297">Gibco™ CTS™ Versene Solution</td> <td style="text-align: center" width="87">100 mL</td> <td style="text-align: center" width="91">0.48 mM</td> <td style="text-align: center" width="159">PBS</td> </tr> <tr> <td style="text-align: center" width="89">13150016</td> <td style="text-align: center" width="297">Gibco™ Cell Dissociation Buffer, enzyme-free, Hanks’ Balanced Salt Solution</td> <td style="text-align: center" width="87">100 mL</td> <td style="text-align: center" width="91">非公開</td> <td style="text-align: center" width="159">HBSS w/o Ca<sup>2+</sup> & Mg<sup>2+</sup><br /> 詳細は非公開</td> </tr> <tr> <td style="text-align: center" width="89">13151014</td> <td style="text-align: center" width="297">Gibco™ Cell Dissociation Buffer,<br /> enzyme-free, PBS</td> <td style="text-align: center" width="87">100 mL</td> <td style="text-align: center" width="91">非公開</td> <td style="text-align: center" width="159">PBS w/o Ca<sup>2+</sup> & Mg<sup>2+</sup><br /> 詳細は非公開</td> </tr> </tbody> </table> <p>Versene solutionは0.48 mMのEDTAが含まれるPBSです。一方、HBSSベースとPBSベースのラインナップがあるCell Dissociation Bufferは、Bufferの組成とEDTAの濃度が細胞解離のために最適化されており、詳細は非公開となっています。</p> <h2><span id="Certificate_of_AnalysisCOA">（補足）分析証明書（Certificate of Analysis、COA）の入手方法</span></h2> <p>Collagenase, Type I, powder（製品番号17100017）のロット2789756の分析証明書を入手する場合を例に入手方法をご紹介します。</p> <p>①当社のWebサイト（https://www.thermofisher.com/jp/ja/home.html）にアクセス<br /> ②検索窓のプルダウンメニューを「分析証明書」に変更する<br /> ③検索窓にロット番号「2789756」を入力してEnterキーまたは検索ボタンを押す<br /> ④「Lot # 2789756」のリンクをクリックする<br /> ⑤分析証明書を確認し、必要に応じてPDFで保存する<br /> ※分析証明書が見つからない場合は<a href="https://www.thermofisher.com/jp/ja/home/global/forms/qa-inquiry-form-jp.html?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener">問合せフォーム</a>からご請求ください。<br /> <img decoding="async" class="aligncenter size-large wp-image-16249" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/01/cell-culture-cell-separation-bid-ts-mbr-24001-02-1024x968.png" alt="" width="760" height="718" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/01/cell-culture-cell-separation-bid-ts-mbr-24001-02-1024x968.png 1024w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/01/cell-culture-cell-separation-bid-ts-mbr-24001-02-300x284.png 300w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/01/cell-culture-cell-separation-bid-ts-mbr-24001-02-768x726.png 768w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2024/01/cell-culture-cell-separation-bid-ts-mbr-24001-02.png 1063w" sizes="(max-width: 760px) 100vw, 760px" /></p> <h2><span id="i-2">まとめ</span></h2> <ul> <li>さまざまな細胞解離試薬があります。臓器や組織の種類や実験目的に応じて使い分けてください。</li> <li>酵素系の試薬は溶解方法や保管条件に注意してください。</li> <li>酵素系の試薬は、製造ロットごとに発行される分析証明書に記載された酵素活性の試験結果を確認して溶解、使用してください。</li> </ul> <h3><span id="3D">【無料ダウンロード】3D細胞培養＆解析ハンドブック</span></h3> <p><a class="btn btn-red" href="https://www.thermofisher.com/jp/ja/home/global/forms/life-science/download-3d-cell-culture-analysis-handbook.html?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener noreferrer">ダウンロードはこちら</a></p> <h3><span id="Gibco">【無料ダウンロード】Gibco多能性幹細胞ハンドブック</span></h3> <p><a class="btn btn-red" href="https://www.thermofisher.com/jp/ja/home/global/forms/stemcell-handbook.html?CID=bid_clb_r04_jp_cp0000_pjt0000_bid00000_0so_blg_cell_culture_cell_separation_bid_ts_mbr_24001_Social_LAB" target="_blank" rel="noopener noreferrer">ダウンロードはこちら</a></p> <p> </p> <p>研究用にのみ使用できます。診断用には使用いただけません。</p> <aside class="mashsb-container mashsb-main "><div class="mashsb_above_buttons">Share this article </div><div class="mashsb-box"><div 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