{"id":2736,"date":"2021-01-08T00:00:00","date_gmt":"2021-01-07T16:00:00","guid":{"rendered":"https:\/\/www.cib.cc\/?p=2736"},"modified":"2021-01-08T00:00:00","modified_gmt":"2021-01-07T16:00:00","slug":"cib-cit-hku%e8%81%94%e5%90%88%e5%bb%ba%e7%ab%8b%e5%8f%8c%e6%8a%a5%e5%91%8a%e5%9f%ba%e5%9b%a0%e6%a0%87%e8%ae%b0%e5%bf%83%e8%82%8c%e7%bb%86%e8%83%9e%e8%b0%b1%e7%b3%bb%e7%89%b9%e5%bc%82%e6%80%a7%e5%9f%ba","status":"publish","type":"post","link":"https:\/\/www.cib.cc\/?p=2736","title":{"rendered":"CIB\/CIT\/HKU\u8054\u5408\u5efa\u7acb\u53cc\u62a5\u544a\u57fa\u56e0\u6807\u8bb0\u5fc3\u808c\u7ec6\u80de\u8c31\u7cfb\u7279\u5f02\u6027\u57fa\u56e0NKX2-5\u7684\u4ebaiPSCs"},"content":{"rendered":"

\u8c03\u67e5\u7814\u7a76\u8868\u660e\uff0c\u6211\u56fd\u6bcf\u5e74\u7531\u4e8e\u5fc3\u810f\u731d\u6b7b\u53d1\u75c5\u7684\u4eba\u5927\u6982\u8d85\u8fc754\u4e07\u4eba\u3002\u9ad8\u5f3a\u5ea6\u7684\u5de5\u4f5c\uff0c\u8fde\u7eed\u8f83\u957f\u7684\u65f6\u95f4\u5de5\u4f5c\u65f6\u957f\u548c\u7ecf\u5e38\u7684\u71ac\u591c\u5bfc\u81f4\u7684\u5fc3\u810f\u65b9\u9762\u7684\u95ee\u9898\u662f\u5f15\u8d77\u5fc3\u810f\u8870\u7aed\u7684\u91cd\u8981\u56e0\u7d20\u30021<\/p>\n

<\/p>\n

\u5fc3\u810f\u662f\u4eba\u4f53\u6700\u91cd\u8981\u7684\u7ef4\u6301\u5668\u5b98\uff0c\u652f\u6301\u5168\u8eab\u8840\u6db2\u5faa\u73af\u3002\u5fc3\u810f\u4e00\u65e6\u9aa4\u505c\uff0c\u5168\u8eab\u7684\u5404\u4e2a\u5668\u5b98\u53ca\u7ec4\u7ec7\u4f1a\u53d1\u751f\u4e0d\u540c\u7a0b\u5ea6\u7684\u7f3a\u8840\uff0c\u7ee7\u800c\u5f15\u8d77\u8eab\u4f53\u5404\u4e2a\u5668\u5b98\u548c\u7ec4\u7ec7\u529f\u80fd\u7684\u969c\u788d\uff0c\u53ca\u4e0d\u540c\u7a0b\u5ea6\u7684\u635f\u4f24\u4ee5\u53ca\u574f\u6b7b\u3002\u5fc3\u810f\u9aa4\u505c\u56db\u5206\u949f\u53ef\u80fd\u5bfc\u81f4\u4e0d\u53ef\u9006\u8f6c\u8111\u635f\u4f24\uff0c\u66f4\u957f\u65f6\u95f4\u5219\u4f1a\u5bfc\u81f4\u8111\u6b7b\u4ea1\u3002<\/p>\n

<\/p>\n

\u5fc3\u808c\u7ec6\u80de\u662f\u5fc3\u810f\u80fd\u591f\u7ef4\u6301\u8282\u5f8b\u6027\u8df3\u52a8\u7684\u57fa\u7840\u5355\u5143\u3002\u5fc3\u808c\u7ec6\u80de\u5c5e\u4e8e\u6210\u4f53\u7ec6\u80de\uff0c\u5728\u4f53\u5916\u65e0\u6cd5\u589e\u6b96\u3002\u5728\u5fc3\u810f\u9886\u57df\u7814\u7a76\u4e2d\uff0c\u5fc3\u808c\u7ec6\u80de\u56e0\u5176\u83b7\u53d6\u9014\u5f84\u6709\u9650\uff0c\u7ec6\u80de\u6279\u6b21\u95f4\u5dee\u5f02\u5927\u800c\u8fdb\u5c55\u7f13\u6162\u3002\u5e38\u7528\u7684\u52a8\u7269\u6a21\u578b\u5982\u5c0f\u9f20\uff0c\u5927\u9f20\u7684\u5fc3\u810f\u6bcf\u5206\u949f\u640f\u52a8\u6b21\u6570\u4e0e\u6b63\u5e38\u4eba\u5dee\u522b\u8f83\u5927\uff0c\u5176\u5fc3\u808c\u7ec6\u80de\u4e0e\u4eba\u5fc3\u808c\u7ec6\u80de\u5728\u751f\u7406\u548c\u529f\u80fd\u4e0a\u4e5f\u6709\u5f88\u5927\u5dee\u522b\uff0c\u8fd9\u5728\u4e00\u5b9a\u7a0b\u5ea6\u4e0a\u9650\u5236\u4e86\u5fc3\u808c\u7ec6\u80de\u5728\u4f53\u5916\u75be\u75c5\u6a21\u578b\u53ca\u75be\u75c5\u673a\u5236\u7814\u7a76\u4e2d\u7684\u5e94\u7528\u3002\u8fd1\u5e74\u6765\u4eba\u591a\u80fd\u5e72\u7ec6\u80de\u901a\u8fc7\u5b9a\u5411\u5206\u5316\u83b7\u5f97\u7684\u5fc3\u808c\u7ec6\u80de\u4e3a\u5fc3\u810f\u75be\u75c5\u7684\u4f53\u5916\u6a21\u578b\uff0c\u75be\u75c5\u673a\u5236\u53ca\u5fc3\u8870\u60a3\u8005\u7684\u7ec6\u80de\u66ff\u4ee3\u7597\u6cd5\u63d0\u4f9b\u4e00\u79cd\u65e0\u9650\u6570\u91cf\u7684\u6765\u6e90\u3002<\/p>\n

<\/p>\n

\u4e3a\u4e86\u63d0\u9ad8\u591a\u80fd\u5e72\u7ec6\u80de\u5206\u5316\u800c\u6765\u7684\u5fc3\u808c\u7ec6\u80de\u7684\u7eaf\u5ea6\uff0c\u5df2\u6709\u591a\u79cd\u65b9\u6cd5\u7528\u4e8e\u5bcc\u96c6\u5fc3\u808c\u7ec6\u80de\u3002\u6bd4\u5982\uff0c\u5229\u7528\u5fc3\u808c\u7ec6\u80de\u7279\u6709\u7684\u4ee3\u8c22\u7279\u70b9\uff0c\u4f7f\u7528Glucose-Depleted and Lactate-Enriched CultureConditions\uff0c\u6216\u662f\u5229\u7528GFP\u6807\u8bb0\u5fc3\u808c\u7ec6\u80de\u524d\u4f53\u7279\u5f02\u6027\u57fa\u56e0NKX2-5\uff0c\u7136\u540e\u901a\u8fc7\u5206\u9009\u7684\u65b9\u5f0f\u83b7\u5f97\u9ad8\u7eaf\u5ea6\u7684\u5fc3\u808c\u7ec6\u80de\u30022<\/p>\n

<\/p>\n

\u4e3a\u4e86\u4f7f\u5fc3\u808c\u7ec6\u80de\u66f4\u5bb9\u6613\u7eaf\u5316\uff0c\u907f\u514d\u6d41\u5f0f\u5206\u9009\u6240\u9700\u8981\u7684\u6d88\u5316\u8fc7\u7a0b\u53ca\u8fc7\u673a\u65b9\u9762\u7684\u538b\u529b\uff0c\u540c\u65f6\u4e3a\u4e86\u4f18\u5316\u5fc3\u808c\u7ec6\u80de\u5206\u5316\u7684\u64cd\u4f5c\u89c4\u8303\uff0c\u6211\u4eec\u501f\u52a9\u4e8eTALENs\u5728NKX2-5\u7b2c\u4e00\u4e2a\u5916\u663e\u5b50\u5904\u63d2\u5165\u4e86eGFP\u548cPURO\u6297\u6027\u57fa\u56e0\uff08Pac\uff092\u4e2a\u62a5\u544a\u57fa\u56e0\u30023<\/p>\n

<\/p>\n

eGFP\u548cPac KI\u793a\u610f\u56fe<\/p>\n

\"1610331021137879.jpg\"<\/p>\n

Zhou et al., 2021. Stem Cell Research<\/span><\/p>\n


<\/span><\/p>\n

\u8be5\u7ec6\u80de\u7cfb\u5206\u5316\u4e3a\u5fc3\u808c\u7ec6\u80de\u540e\uff0cday 8\u5de6\u53f3NKX2-5\u5f00\u59cb\u8868\u8fbe\uff0ceGFP\u5f00\u59cb\u540c\u6b65\u8868\u8fbe\uff0c\u5e76\u5728day 11-12\u5de6\u53f3\u5f00\u59cb\u7a33\u5b9a\uff0c\u968f\u540e\u4e00\u76f4\u5728\u8df3\u52a8\u7684\u5fc3\u808c\u7ec6\u80de\u4e2d\u8868\u8fbe\u3002\u6211\u4eec\u5728day 8\u5f00\u59cb\u4f7f\u7528Puromycin\u5904\u7406\u7ec6\u80de\uff0c8\u5929\u540e\uff0c\u5b58\u6d3b\u7684\u7ec6\u80de\u57fa\u672c\u4e0a\u5747\u4e3aGFP\u9633\u6027\u7684\u7ec6\u80de\u3002<\/p>\n

\"NKX2-5<\/p>\n

NKX2-5 GFP-Puro\/wt iPSC\u6765\u6e90\u7684\u5fc3\u808c\u7ec6\u80de (Zhou et al., 2021. Stem Cell Research)<\/span><\/p>\n


<\/span><\/p>\n

\u7531\u4e8e\u643a\u5e26GFP\u548cPuromycin\u6297\u6027\u53cc\u62a5\u544a\u57fa\u56e0\uff0c\u8be5\u7ec6\u80de\u7cfb\u53ef\u7528\u4e8e\u4eba\u591a\u80fd\u5e72\u7ec6\u80de\u5411\u5fc3\u808c\u7ec6\u80de\u5b9a\u5411\u5206\u5316Protocol\u7684\u4f18\u5316\uff0c\u5927\u91cf\u751f\u4ea7\u5de5\u4e1a\u5316\u9700\u6c42\u7684\u9ad8\u7eaf\u5ea6\u5fc3\u808c\u7ec6\u80de\uff0c\u5fc3\u810f\u6bd2\u6027\u6d4b\u8bd5\u7b49\u3002\u4e09\u542f\u751f\u7269\u5df2\u83b7\u5f97\u5229\u7528\u8be5\u65b9\u6cd5\u751f\u4ea7\u9ad8\u7eaf\u5ea6\u5fc3\u808c\u7ec6\u80de\u7684\u4e13\u5229\uff08ZL201610149395.0\uff0c\u4e00\u79cd\u83b7\u5f97\u9ad8\u7eaf\u5ea6\u5fc3\u808c\u7ec6\u80de\u7684\u65b9\u6cd5\uff09\u3002<\/p>\n

<\/p>\n

\u8be5\u7ec6\u80de\u7cfb\u7531\u4e09\u542f\u751f\u7269\uff08CIB\uff09\u5efa\u7acb\uff0c\u540e\u7eed\u9274\u5b9a\u7531CIB\u3001\u4e09\u542f\u836f\u7269\uff08CIT\uff09\u8054\u5408\u9999\u6e2f\u5927\u5b66\uff08HKU\uff09\u674e\u5609\u8bda\u533b\u5b66\u9662\u5fc3\u5185\u79d1\u5171\u540c\u5b8c\u6210\u3002\u76f8\u5173\u6587\u7ae0\u4ee5Lab Resources\u65b9\u5f0f\u53d1\u8868\u5728\u6700\u65b0\u4e00\u671f\u7684Stem Cell Research (https:\/\/doi.org\/10.1016\/j.scr.2020.102120).<\/p>\n


<\/span>\u76f8\u5173\u4ea7\u54c1<\/p>\n\n\n\n\n\n
\u8d27\u53f7<\/td>\n\u4ea7\u54c1<\/td>\n\u5907\u6ce8<\/td>\n<\/tr>\n
iCar100<\/td>\n\n

NKX2-5GFP-Puro\/wt iPSC\u6765\u6e90\u5fc3\u808c\u7ec6\u80de<\/p>\n<\/td>\n

8*105\u7ec6\u80de\/\u7ba1\uff0c\u542b\u590d\u82cf\u53ca\u7ef4\u6301\u57f9\u517b\u57fa<\/td>\n<\/tr>\n
iCar002<\/td>\n\u5fc3\u808c\u7ec6\u80de\u7ef4\u6301\u57f9\u517b\u57fa\uff08100ml\uff09<\/td>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

<\/p>\n

\n

\u53c2\u8003\u6587\u732e<\/strong><\/p>\n<\/section>\n

\n


<\/p>\n<\/section>\n

\n

1.https:\/\/www.medsci.cn\/article\/show_article.do?id=40d3898e58
<\/span><\/p>\n

2. Elliott David A,Braam Stefan R,Koutsis Katerina et al. NKX2-5(eGFP\/w) hESCs for isolation of human cardiac progenitors and cardiomyocytes.[J] .Nat Methods, 2011, 8: 1037-40.<\/span><\/p>\n

3. M. Zhou, R. Wei, Y. Jiang, J. Fu, Y. Liu, B. Yang, B. Yu, Y. Lin, X. Ran, W-H. Lai,M. Chu, Y. Hu, J. Yang, H-F. Tse, Generation of a Human iPSC Line GIBHi002-A-2 with Dual-Reporter for NKX2-5 Using TALENs, Stem Cell Research (2020) , doi: https:\/\/doi.org\/10.1016\/j.scr.2020.102120<\/span><\/p>\n<\/section>\n

<\/p>\n","protected":false},"excerpt":{"rendered":"

\u8c03\u67e5\u7814\u7a76\u8868\u660e\uff0c\u6211\u56fd\u6bcf\u5e74\u7531\u4e8e\u5fc3\u810f\u731d\u6b7b\u53d1\u75c5\u7684\u4eba\u5927\u6982\u8d85\u8fc754\u4e07\u4eba\u3002\u9ad8\u5f3a\u5ea6\u7684\u5de5\u4f5c\uff0c\u8fde\u7eed\u8f83\u957f\u7684\u65f6\u95f4\u5de5\u4f5c\u65f6\u957f\u548c\u7ecf\u5e38\u7684\u71ac\u591c\u5bfc […]<\/p>\n","protected":false},"author":1,"featured_media":2737,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[38],"tags":[],"class_list":["post-2736","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-news"],"blocksy_meta":[],"_links":{"self":[{"href":"https:\/\/www.cib.cc\/index.php?rest_route=\/wp\/v2\/posts\/2736","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.cib.cc\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.cib.cc\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.cib.cc\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.cib.cc\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=2736"}],"version-history":[{"count":0,"href":"https:\/\/www.cib.cc\/index.php?rest_route=\/wp\/v2\/posts\/2736\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.cib.cc\/index.php?rest_route=\/wp\/v2\/media\/2737"}],"wp:attachment":[{"href":"https:\/\/www.cib.cc\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=2736"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.cib.cc\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=2736"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.cib.cc\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=2736"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}