\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u3002\u8be5\u7ec6\u80de\u7f3a\u4e4f HGPRT\uff08HPRT\uff09\uff0c\u5e76\u4e14\u5bf9 0.06 mM \u7684 6-\u786b\u9e1f\u560c\u5464\u6709\u6297\u6027\u3002\u5f53\u5728\u9972\u517b\u5c42\uff08\u80da\u80ce\u6210\u7ea4\u7ef4\u7ec6\u80de\u6216 STO \u7ec6\u80de\uff09\u4e0a\u57f9\u517b\u65f6\uff0c\u7ec6\u80de\u4fdd\u6301\u672a\u5206\u5316\u72b6\u6001\u3002\u5728\u6ca1\u6709\u9972\u517b\u5c42\u7684\u60c5\u51b5\u4e0b\uff0c\u7ec6\u80de\u81ea\u53d1\u5206\u5316\u5e76\u5f62\u6210\u80da\u80ce\u7ed3\u6784\u3002\u5f53\u6ce8\u5165\u80da\u6ce1\u4e2d\u65f6\uff0c\u7ec6\u80de\u53ef\u4ee5\u8fdb\u5165\u751f\u6b96\u7cfb\u3002\u5728\u5e38\u89c4\u7684\u5206\u5b50\u57fa\u56e0\u4fee\u9970\u6280\u672f\u4e4b\u540e\uff0c\u5b83\u4eec\u53ef\u7528\u4e8e\u91cd\u6784\u5c0f\u9f20\u80da\u80ce\u3002\u57f9\u517b\u65f6\u9700\u4f7f\u7528\u6606\u660e\u767dMEF\u4f5c\u4e3a\u9972\u517b\u5c42\u7ec6\u80de\u3002<\/p>\n
\u7ec6\u80de\u7279\u6027<\/p>\n
1\uff09 \u6765\u6e90\uff1a\u5c0f\u9f20\uff0c129\/Ola\u54c1\u7cfb\uff0c\u80da\u80ce\u5185\u7ec6\u80de\u56e2<\/p>\n
2\uff09 \u5f62\u6001\uff1a\u7403\u5f62\u514b\u9686 \u8d34\u58c1\u751f\u957f<\/p>\n
3\uff09 \u542b\u91cf\uff1a>1×10^6 \u7ec6\u80de\u6570<\/p>\n
4\uff09 \u89c4\u683c\uff1a1mL\u51bb\u5b58\u7ba1\u5305\u88c5<\/p>\n
5\uff09 \u7528\u9014\uff1a\u4ec5\u4f9b\u79d1\u7814\u4f7f\u7528<\/p>\n
\u8fd0\u8f93\u548c\u4fdd\u5b58<\/p>\n
\u5e72\u51b0\u8fd0\u8f93\uff1a1mL\u51bb\u5b58\u7ba1\u5305\u88c5\u5e72\u51b0\u8fd0\u8f93\uff0c\u6536\u5230\u540e-80\u5ea6\u51b0\u7bb1\u4fdd\u5b58\u8fc7\u591c\u540e\u8f6c\u5165\u6db2\u6c2e\u6216\u76f4\u63a5\u590d\u82cf\uff0c\u82e5\u53d1\u73b0\u5e72\u51b0\u5df2\u6325\u53d1\u5e72\u51c0\u3001\u51bb\u5b58\u7ba1\u74f6\u76d6\u8131\u843d\u3001\u7834\u635f\u53ca\u7ec6\u80de\u6709\u6c61\u67d3\uff0c\u8bf7\u7acb\u5373\u4e0e\u6211\u4eec\u8054\u7cfb\u3002<\/p>\n
\u4e00\uff0e\u57f9\u517b\u57fa\u53ca\u57f9\u517b\u51bb\u5b58\u6761\u4ef6\u51c6\u5907<\/p>\n
1\uff09\u51c6\u5907DMEM \u57fa\u7840\u57f9\u517b\u57fa (iCell-0001) 81.5%<\/p>\n
\u4f18\u8d28\u80ce\u725b\u8840\u6e05 \uff08iCell-0500\uff09 15 %<\/p>\n
GlutaMAX-1\u8c37\u6c28\u9170\u80fa ( iCell-0900) 1 %<\/p>\n
MEM NEAA\u975e\u5fc5\u9700\u6c28\u57fa\u9178 (iCell-01000) 1%<\/p>\n
P\/S\u9752\u9709\u7d20-\u94fe\u9709\u7d20 \uff08iCell-15140-122\uff09 1%<\/p>\n
\u03b2-\u5def\u57fa\u4e59\u9187 (iCell-8211) 1.25 mL<\/p>\n
LIF (iCell-50756) 10ng\/mL<\/p>\n
\u6ce8\u610f\uff1a<\/p>\n
1\u3001\u5efa\u8bae\u4f7f\u7528KM- MEF\u4f5c\u4e3a\u9972\u517b\u5c42\u7ec6\u80de\u3002<\/p>\n
2\u3001\u5728\u94fa\u80da\u80ce\u5e72\u7ec6\u80de\u524d\uff0c\u9700\u5bf9MEF\u8fdb\u884c\u5904\u7406\uff0c\u5177\u4f53\u5904\u7406\u6b65\u9aa4\u89c1\u4e0b\u6587\u4e1d\u88c2\u9709\u7d20\u706d\u6d3bMEF\u3002<\/p>\n
3\u3001\u8be5\u7ec6\u80de\u5efa\u8bae\u51bb\u5b58\u5e72\u51b0\u53d1\u8d27\uff0c\u4e0d\u9002\u5408\u957f\u65f6\u95f4\u6d3b\u7ec6\u80de\u8fd0\u8f93\uff0c\u4f1a\u5f71\u54cd\u7ec6\u80de\u6d3b\u529b\u3002<\/p>\n
2\uff09\u57f9\u517b\u6761\u4ef6\uff1a \u6c14\u76f8\uff1a\u7a7a\u6c14\uff0c95%\uff1b\u4e8c\u6c27\u5316\u78b3\uff0c5%\u3002 \u6e29\u5ea6\uff1a37\u2103\uff0c\u57f9\u517b\u7bb1\u6e7f\u5ea6\u4e3a70%-80%\u3002<\/p>\n
3\uff09\u51bb\u5b58\u6db2\uff1a\u5b8c\u5168\u57f9\u517b\u6db2 60%%\uff0cFBS 30%%\uff0cDMSO 10%%\u73b0\u7528\u73b0\u914d\u3002<\/p>\n
\u6211\u5e93\u51bb\u5b58\u65f6\uff0c\u4f53\u79ef\u4e3a 500 \u03bcl\uff0c\u9884\u671f\u5b58\u6d3b\u7387 70%\uff0c\u6bcf\u652f\u51bb\u5b58\u7ba1\u7684\u7ec6\u80de\u590d\u82cf\uff0c3 \u81f34 \u5929\u540e\uff0c\u4f1a\u5f62\u6210 30 \u81f3 40 \u4e2a\u514b\u9686\uff0c\u5efa\u8bae\u590d\u82cf\u81f3 1 \u4e2a T25 \u57f9\u517b\u74f6\u4e2d\u3002<\/p>\n
\u4e8c\uff1a\u7ec6\u80de\u5904\u7406<\/p>\n
\u4e1d\u88c2\u9709\u7d20\u706d\u6d3bMEF<\/p>\n
\u5f85MEF\u7ec6\u80de\u5bc6\u5ea6\u8fbe\u523080%\u4ee5\u4e0a\uff0c\u52a0\u5165\u542b\u4e1d\u88c2\u9709\u7d20\uff0815\u03bcg\/mL\uff09\u7684MEF\u5b8c\u5168\u57f9\u517b\u6db2\uff0c\u7f6e\u4e8e37\u57f9\u517b\u7bb1\u4e2d\u5b75\u80b22.5h\uff0c2.5h\u540e\uff0c\u5f03\u6389\u57f9\u517b\u6db2\uff0cPBS\u6e05\u6d171-2\u904d\uff0c\u52a0\u5165MEF\u57f9\u517b\u57fa\uff0c\u5907\u7528\uff0c\u5f53\u59292h\u540e\u6216\u8005\u7b2c\u4e8c\u5929\u53ef\u4ee5\u4f20\u5165\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u3002<\/p>\n
\u590d\u82cf\uff1a<\/p>\n
1.\u5c06\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u51bb\u5b58\u7ba1\u4ece\u6db2\u6c2e\u4e2d\u53d6\u51fa\uff0c\u7f6e\u4e8e 37\u2103\u6c34\u6d74\u4e2d\u4f7f\u4e4b\u8fc5\u901f\u878d\u89e3\uff0c\u53d6\u51fa\u540e\u62ff\u5230\u8d85\u51c0\u53f0\u5185\u7528 75%\u9152\u7cbe\u64e6\u62ed\u51bb\u5b58\u7ba1\u65cb\u53e3\u5904\u53ca\u5916\u58c1\uff0c\u9632\u6b62\u6c61\u67d3\u3002<\/p>\n
2. \u5c06\u51bb\u5b58\u7ba1\u5185\u7ec6\u80de\u60ac\u6db2\u8f6c\u79fb\u81f3\u542b 3-4 ml \u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u5b8c\u5168\u57f9\u517b\u6db2\u7684 15ml \u79bb\u5fc3\u7ba1\u5185\uff0c\u4ee5 1000 rpm\uff0c\u79bb\u5fc3 5 min\u3002<\/p>\n
3. \u79bb\u5fc3\u540e\u5c06\u4e0a\u6e05\u6db2\u5438\u9664\uff0c\u53e6\u52a0\u5165\u65b0\u9c9c\u7684\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u5b8c\u5168\u57f9\u517b\u6db2 2 ml\uff0c\u5439\u6253\u60ac\u6d6e\u3002<\/p>\n
4. \u91cd\u590d\u5439\u6253\uff0c\u5236\u6210\u5355\u7ec6\u80de\u60ac\u6db2\uff0c\u5c3d\u91cf\u907f\u514d\u6c14\u6ce1\u3002<\/p>\n
5. \u8f6c\u79fb\u81f3 1 \u4e2a\u5df2\u7ecf\u94fa\u597d MEF \u7ec6\u80de\u7684 T25 \u57f9\u517b\u74f6\u4e2d\u57f9\u517b\u3002<\/p>\n
6. \u6bcf\u5929\u66f4\u6362\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u5b8c\u5168\u57f9\u517b\u6db2\u3002<\/p>\n
\u4f20\u4ee3\uff1a<\/p>\n
1. \u4e00\u822c\u5728\u590d\u82cf\u540e\u7b2c 2-3 \u5929\u4f20\u4ee3\uff0c\u89c6\u514b\u9686\u5927\u5c0f\u548c\u5bc6\u5ea6\u800c\u5b9a<\/p>\n
2. \u5438\u9664\u5e9f\u6db2\u3002<\/p>\n
3. \u7528 PBS\uff08\u4e0d\u542b\u9499\u9541\u79bb\u5b50\uff09\u8f7b\u8f7b\u51b2\u6d17\u4e00\u904d\u3002<\/p>\n
4. \u52a0\u5165 1.0 ml \u7684 0.25%\u80f0\u9176\uff08\u542b EDTA\uff09\u81f3\u57f9\u517b\u74f6\uff0c\u8f7b\u8f7b\u6643\u52a8\uff0c\u4f7f\u80f0\u9176\u8986\u76d6\u5e95\u9762\uff0c\u7f6e\u4e8e 37\u2103\u57f9\u517b\u7bb1\u5185\u6d88\u5316\u7ec6\u80de\u3002<\/p>\n
5. \u5728\u663e\u5fae\u955c\u4e0b\u89c2\u5bdf\uff0c\u76f4\u81f3\u7ec6\u80de\u5c42\u5168\u90e8\u8131\u843d\uff08\u4e00\u822c\u9700\u8981 1-2 min\uff09\u3002<\/p>\n
6. \u52a0 2 ml \u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u5b8c\u5168\u57f9\u517b\u6db2\u7ec8\u6b62\u6d88\u5316\u3002<\/p>\n
7. \u4ee5 1000 rpm\uff0c\u79bb\u5fc3 5 min\uff0c\u5f03\u4e0a\u6e05\u3002<\/p>\n
8. \u52a0\u5165\u7ea6 1 ml \u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u5b8c\u5168\u57f9\u517b\u6db2\uff0c\u591a\u6b21\u8f7b\u8f7b\u5439\u6253\u7ec6\u80de\uff0c \u5236\u6210\u5355\u7ec6\u80de\u60ac\u6db2\u3002<\/p>\n
9. \u52a0\u5165\u8db3\u91cf\u7684\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u5b8c\u5168\u57f9\u517b\u6db2\uff0c\u5439\u6253\u6df7\u5300\uff0c\u7ec6\u80de\u60ac\u6db2\u5206\u88c5\u5230\u94fa\u597d MEF \u7ec6\u80de\u7684 T25 \u57f9\u517b\u74f6\u4e2d\u3002\u4e00\u822c\u5730\uff0c\u4e00\u4e2a T25 \u57f9\u517b\u74f6\u52a0\u5165 5-6 ml \u57f9\u517b\u6db2\u3002\u653e\u5165 37\u2103\u57f9\u517b\u7bb1\u5185\u57f9\u517b\u3002\u6bcf\u5929\u6362\u6db2\u3002<\/p>\n
\u4f20\u4ee3\u6bd4\u4f8b\uff1a1:4-1:7<\/p>\n
\u51bb\u5b58\uff1a<\/p>\n
1.\u6309\u4f20\u4ee3\u7684\u65b9\u6cd5\u5c06\u7ec6\u80de\u6d88\u5316\u4e0b\u6765\uff0c\u5236\u6210\u7ec6\u80de\u60ac\u6db2\u3002<\/p>\n
2. \u4ee5 1000 rpm\uff0c\u79bb\u5fc3 5 min\uff0c\u5f03\u4e0a\u6e05\uff0c\u9010\u6ef4\u52a0\u5165\u5df2\u7ecf\u9884\u51b7\u7684\u51bb\u5b58\u6db2\uff0c\u60ac\u6d6e\u7ec6\u80de\u3002<\/p>\n
3. \u6309\u6bcf\u652f\u5b58\u7ba1\u5185\u52a0\u5165 500 ml \u7ec6\u80de\u60ac\u6db2\u5206\u88c5\u5230\u51bb\u5b58\u7ba1\u5185\uff0c\u6807\u8bb0\u7ec6\u80de\u540d\u79f0\u3001\u4ee3\u6570\u3001\u51bb\u5b58\u65e5\u671f\u7b49\u57fa\u672c\u4fe1\u606f\u3002<\/p>\n
4\uff0e\u5c06\u51bb\u5b58\u7ba1\u7f6e\u4e8e\u7a0b\u5e8f\u964d\u6e29\u76d2\u5185\uff0c-80\u2103\u8fc7\u591c\uff0c\u8f6c\u5165\u6db2\u6c2e\u3002<\/p>\n
\u51bb\u5b58\u6db2\u914d\u65b9\uff1a<\/p>\n
\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u5b8c\u5168\u57f9\u517b\u6db2 60%\uff0cES \u7ea7 FBS 30%\uff0cDMSO 10%<\/p>\n
\u9644\uff1a\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u4e0e MEF \u7ec6\u80de\u5206\u79bb\u7684\u7b80\u6613\u65b9\u6cd5\uff08\u5dee\u901f\u8d34\u58c1\u6cd5\uff09\uff1a<\/p>\n
1.\u57f9\u517b\u4e2d\u7684\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\uff0c\u6309\u4f20\u4ee3\u7684\u64cd\u4f5c\u65b9\u6cd5\u5c06\u7ec6\u80de\u7528\u80f0\u9176\u6d88\u5316\u5e76\u5439\u6253\u6210\u5355\u7ec6\u80de\u60ac\u6db2\u540e\uff0c\u52a0\u5165\u9002\u91cf\u7684\u63d0\u524d\u6e29\u80b2\u597d\u7684\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u5b8c\u5168\u57f9\u517b\u6db2\u91cd\u60ac\u7ec6\u80de\uff0c \u5439\u6253\u6df7\u5300\uff0c\u7ec6\u80de\u60ac\u6db2\u5206\u88c5\u5230\u65e0 MEF \u7ec6\u80de\u7684\u57f9\u517b\u76bf\u6216\u57f9\u517b\u74f6\uff08\u4e00\u822c\u5730\uff0c\u4e00\u4e2aT25 \u57f9\u517b\u74f6\u4e2d\u57f9\u517b\u7684\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\uff0c\u5728\u4e00\u4e2a 10 cm \u57f9\u517b\u76bf\u4e2d\u8fdb\u884c\u5dee\u901f\u8d34\u58c1\u3002\u4e0d\u8981\u4e8b\u5148\u94fa\u660e\u80f6\uff0c\u5982\u94fa\u660e\u80f6\u5219\u7ec6\u80de\u8d34\u58c1\u901f\u5ea6\u8fc7\u5feb\u65e0\u6cd5\u6709\u6548\u5206\u79bb\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u4e0e MEF \u7ec6\u80de\uff09\u4e2d\u3002<\/p>\n
2. \u57f9\u517b\u76bf\u6216\u57f9\u517b\u74f6\u7f6e\u4e8e 37\u00b0C \u57f9\u517b\u7bb1\u5185\u9759\u7f6e 1 \u5c0f\u65f6\u3002<\/p>\n
3. 1 \u5c0f\u65f6\u540e\uff0c\u7edd\u5927\u90e8\u5206 MEF \u7ec6\u80de\u5df2\u8d34\u5728\u57f9\u517b\u76bf\u6216\u57f9\u517b\u74f6\u5e95\u9762\uff0c\u800c\u5927\u90e8\u5206\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u4ecd\u7136\u60ac\u6d6e\u5728\u57f9\u517b\u6db2\u4e2d\u3002\u6536\u53d6\u57f9\u517b\u6db2\uff0c\u8fdb\u884c\u540e\u7eed\u5b9e\u9a8c\u3002<\/p>\n","protected":false},"excerpt":{"rendered":"
\u5c0f\u9f20\u80da\u80ce\u5e72\u7ec6\u80de\u3002\u8be5\u7ec6\u80de\u7f3a\u4e4f HGPRT\uff08HPRT\uff09\uff0c\u5e76\u4e14\u5bf9 0.06 mM \u7684 6-\u786b\u9e1f\u560c\u5464\u6709\u6297\u6027\u3002\u5f53\u5728\u9972\u517b\u5c42 […]<\/p>\n","protected":false},"featured_media":591,"comment_status":"open","ping_status":"closed","template":"","meta":[],"product_brand":[],"product_cat":[27],"product_tag":[],"class_list":["post-589","product","type-product","status-publish","has-post-thumbnail","product_cat-mouse-cell-lines","first","instock","shipping-taxable","purchasable","product-type-simple"],"_links":{"self":[{"href":"http:\/\/www.imorock.com\/index.php\/wp-json\/wp\/v2\/product\/589","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/www.imorock.com\/index.php\/wp-json\/wp\/v2\/product"}],"about":[{"href":"http:\/\/www.imorock.com\/index.php\/wp-json\/wp\/v2\/types\/product"}],"replies":[{"embeddable":true,"href":"http:\/\/www.imorock.com\/index.php\/wp-json\/wp\/v2\/comments?post=589"}],"wp:featuredmedia":[{"embeddable":true,"href":"http:\/\/www.imorock.com\/index.php\/wp-json\/wp\/v2\/media\/591"}],"wp:attachment":[{"href":"http:\/\/www.imorock.com\/index.php\/wp-json\/wp\/v2\/media?parent=589"}],"wp:term":[{"taxonomy":"product_brand","embeddable":true,"href":"http:\/\/www.imorock.com\/index.php\/wp-json\/wp\/v2\/product_brand?post=589"},{"taxonomy":"product_cat","embeddable":true,"href":"http:\/\/www.imorock.com\/index.php\/wp-json\/wp\/v2\/product_cat?post=589"},{"taxonomy":"product_tag","embeddable":true,"href":"http:\/\/www.imorock.com\/index.php\/wp-json\/wp\/v2\/product_tag?post=589"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}