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70%-80%\uff0c\u786e\u4fdd\u5230\u8fbe\u540e\u53ef\u76f4\u63a5\u7528\u4e8e\u5b9e\u9a8c\u6216\u4f20\u4ee3\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u652f\u539f\u4f53\u68c0\u6d4b\uff1a\u65e0\uff08\u6bcf\u6279\u6b21\u7ec6\u80de\u901a\u8fc7 PCR \u6cd5\u68c0\u6d4b\u652f\u539f\u4f53\uff08\u8986\u76d6\u80ba\u708e\u652f\u539f\u4f53\u3001\u4eba\u578b\u652f\u539f\u4f53\u3001\u89e3\u8132\u652f\u539f\u4f53\uff09\uff0c\u7ed3\u679c\u4e3a\u9634\u6027\uff0c\u907f\u514d\u652f\u539f\u4f53\u6c61\u67d3\u5f71\u54cd\u7ec6\u80de\u589e\u6b96\u53ca\u4f53\u5185\u79fb\u690d\u6210\u7624\u80fd\u529b\uff0c\u4fdd\u969c\u4e0e\u5176\u4ed6\u80c3\u764c\u7ec6\u80de\u5e73\u884c\u5b9e\u9a8c\u7684\u53ef\u9760\u6027\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u57f9\u517b\u57fa\uff1a90% DMEM+10% FBS+PS\uff08\u9752\u9709\u7d20 &#8211; \u94fe\u9709\u7d20\uff09\uff08\u6838\u5fc3\u7279\u8272\uff1a\u2460\u91c7\u7528 DMEM \u57f9\u517b\u57fa\uff0c\u533a\u522b\u4e8e NCI-N87 \u7684 RPMI-1640\u3001MKN-45 \u7684 RPMI-1640\uff1bDMEM \u542b\u9ad8\u7cd6\uff084.5g\/L\uff09\u53ca\u4e30\u5bcc\u6c28\u57fa\u9178\uff0c\u9002\u914d\u80c3\u817a\u764c\u8f6c\u79fb\u7076\u7684\u9ad8\u4ee3\u8c22\u9700\u6c42\uff0c\u7ef4\u6301\u5176\u817a\u7ba1\u6837\u5206\u5316\u53ca\u4f53\u5185\u6210\u7624\u80fd\u529b\uff1b\u246110% FBS \u9700\u9009\u62e9\u4f4e\u5185\u6bd2\u7d20\u3001\u6279\u6b21\u95f4\u5dee\u5f02\u5c0f\u7684\u4f18\u8d28\u80ce\u725b\u8840\u6e05\uff0c\u907f\u514d\u8840\u6e05\u4e2d\u5206\u5316\u6291\u5236\u56e0\u5b50\u5f71\u54cd\u817a\u7ba1\u6837\u5f62\u6001\uff1b\u2462PS \u7ec8\u6d53\u5ea6\u4e3a 100U\/mL \u9752\u9709\u7d20\u3001100\u03bcg\/mL \u94fe\u9709\u7d20\uff0c\u6709\u6548\u9884\u9632\u7ec6\u83cc\u6c61\u67d3\uff0c\u4e0d\u5f71\u54cd\u7ec6\u80de\u8f6c\u79fb\u7279\u6027\u53ca\u4f53\u5185\u79fb\u690d\u6548\u679c\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u57f9\u517b\u6761\u4ef6\uff1a\u6c14\u76f8\uff1a95% \u7a7a\u6c14 + 5% \u4e8c\u6c27\u5316\u78b3\uff1b\u6e29\u5ea6\uff1a37\u2103\uff08\u57f9\u517b\u7bb1\u6e7f\u5ea6\u7ef4\u6301\u5728 70%-80%\uff0c\u5b9a\u671f\u66f4\u6362\u6c34\u76d8\uff0c\u9632\u6b62\u57f9\u517b\u57fa\u84b8\u53d1\u5bfc\u81f4\u6e17\u900f\u538b\u5347\u9ad8\uff1b\u4e0e NCI-N87\u3001MKN-45\u3001GES-1 \u7b49\u6240\u6709\u5df2\u68b3\u7406\u7ec6\u80de\u7684\u57f9\u517b\u6761\u4ef6\u5b8c\u5168\u4e00\u81f4\uff0c\u65e0\u9700\u7279\u6b8a\u8c03\u6574\uff0c\u4fbf\u4e8e\u591a\u7ec6\u80de\u682a\u5e76\u884c\u5b9e\u9a8c\uff0c\u5c24\u5176\u9002\u5408\u5f00\u5c55 \u201c\u4e0d\u540c\u4e9a\u578b\u80c3\u764c\u201d \u7684\u4f53\u5185\u79fb\u690d\u5bf9\u6bd4\u5b9e\u9a8c\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u51bb\u5b58\u6761\u4ef6\uff1a\u65e0\u8840\u6e05\u51bb\u5b58\u6db2\uff0c\u6db2\u6c2e\u50a8\u5b58\uff08\u51bb\u5b58\u64cd\u4f5c\u9002\u914d\u817a\u764c\u8f6c\u79fb\u7279\u6027\uff1a\u2460\u9009\u62e9\u5bf9\u6570\u751f\u957f\u671f\u7ec6\u80de\uff08\u63a5\u79cd\u540e 48-72 \u5c0f\u65f6\uff0c\u817a\u7ba1\u6837\u5f62\u6001\u660e\u663e\uff09\uff0c\u7528 0.25% \u80f0\u9176 &#8211; EDTA \u5b75\u80b2 2-3 \u5206\u949f\uff0c\u955c\u4e0b\u89c1\u7ec6\u80de\u95f4\u9699\u589e\u5927\u3001\u817a\u7ba1\u7ed3\u6784\u677e\u6563\u5373\u53ef\u7ec8\u6b62\uff0c\u8f7b\u67d4\u5439\u6253\u907f\u514d\u7834\u574f\u817a\u7ba1\u6837\u7ed3\u6784\uff1b\u24611000rpm \u79bb\u5fc3 5 \u5206\u949f\u6536\u96c6\u7ec6\u80de\uff0c\u7528\u9884\u51b7\u7684\u65e0\u8840\u6e05\u51bb\u5b58\u6db2\u91cd\u60ac\uff0c\u8c03\u6574\u6d53\u5ea6\u81f3 1.5\u00d710\u2076cells\/mL\uff1b\u2462\u51bb\u5b58\u6d41\u7a0b\uff1a4\u2103\u5e73\u8861 30 \u5206\u949f\u2192-20\u2103\u653e\u7f6e 2 \u5c0f\u65f6\uff08\u7a0b\u5e8f\u964d\u6e29\u76d2\uff09\u2192-80\u2103\u8fc7\u591c\u2192\u6db2\u6c2e\u4fdd\u5b58\uff1b\u590d\u82cf\u65f6 37\u2103\u6c34\u6d74 1 \u5206\u949f\u878d\u5316\uff0c\u79bb\u5fc3\u540e\u7528\u9884\u70ed\u7684 DMEM \u57f9\u517b\u57fa\u91cd\u60ac\uff0c\u76f4\u63a5\u63a5\u79cd\uff0c\u51cf\u5c11\u5bf9\u8f6c\u79fb\u76f8\u5173\u8868\u578b\u7684\u5f71\u54cd\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7ec6\u80de\u8d27\u671f\uff1a\u73b0\u8d27\uff0c1 \u5468\u5de6\u53f3\uff08\u5e93\u5b58\u7ec6\u80de\u5747\u7ecf\u8fc7 DMEM \u57f9\u517b\u57fa\u9002\u5e94\u6027\u57f9\u517b\u3001\u817a\u7ba1\u6837\u5f62\u6001\u9a8c\u8bc1\u53ca\u4f53\u5185\u6210\u7624\u9884\u5b9e\u9a8c\uff08\u5982 ICR \u5c0f\u9f20\u76ae\u4e0b\u63a5\u79cd\u9a8c\u8bc1\uff09\uff0c\u786e\u4fdd\u6838\u5fc3\u7279\u6027\u7a33\u5b9a\uff1b\u4e0b\u5355\u540e 48 \u5c0f\u65f6\u5185\u53ef\u5b89\u6392\u53d1\u8d27\uff0c\u53ef\u4e0e NCI-N87\u3001MKN-45 \u540c\u6b65\u4e0b\u5355\uff0c\u4fdd\u969c \u201c\u4e0d\u540c\u6027\u522b \/ \u4e9a\u578b\u201d \u5e73\u884c\u5b9e\u9a8c\u7684\u65f6\u95f4\u4e00\u81f4\u6027\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u8fd0\u8f93\u65b9\u5f0f\uff1a\u590d\u82cf\u53d1\u8d27\uff08T25 \u74f6\u514d\u8fd0\u8f93\u8d39\u7528\uff0c\u91c7\u7528\u9632\u9707\u6052\u6e29\u6ce1\u6cab\u7bb1\uff0c\u5185\u7f6e 2-8\u2103\u51b0\u888b\u53ca\u6e29\u5ea6\u76d1\u6d4b\u5361\uff0c\u8fd0\u8f93\u8fc7\u7a0b\u4e2d\u907f\u514d\u5267\u70c8\u98a0\u7c38\uff0c\u51cf\u5c11\u8d34\u58c1\u7ec6\u80de\u8131\u843d\u53ca\u817a\u7ba1\u6837\u7ed3\u6784\u7834\u574f\uff1b\u5230\u8fbe\u540e\u7acb\u5373\u653e\u5165 37\u2103\u57f9\u517b\u7bb1\uff0c24 \u5c0f\u65f6\u540e\u89c2\u5bdf\u7ec6\u80de\u8d34\u58c1\u60c5\u51b5\uff0c3 \u5929\u5185\u9996\u6b21\u6362\u6db2\uff09\/ \u51bb\u5b58\u53d1\u8d27\uff08\u9700\u52a0\u5e72\u51b0\u8fd0\u8f93\u8d39\u7528\uff0c\u5e72\u51b0\u7528\u91cf\u6309\u8fd0\u8f93\u65f6\u957f\u8ba1\u7b97\uff0c\u56fd\u5185\u8fd0\u8f93\u4e00\u822c\u914d\u5907 500g-1kg \u5e72\u51b0\uff0c\u786e\u4fdd\u5230\u8fbe\u65f6\u5e72\u51b0\u5269\u4f59\u91cf\u226550%\uff0c\u7ef4\u6301\u7ec6\u80de\u51bb\u5b58\u72b6\u6001\uff09 \u987a\u4e30\u5feb\u9012\uff08\u4f18\u5148\u9009\u62e9\u6b21\u65e5\u8fbe\u670d\u52a1\uff0c\u7f29\u77ed\u8fd0\u8f93\u65f6\u95f4\uff0c\u964d\u4f4e\u7ec6\u80de\u6d3b\u6027\u635f\u5931\u53ca\u8f6c\u79fb\u7279\u6027\u6539\u53d8\u98ce\u9669\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u4f9b\u5e94\u9650\u5236\uff1a\u4ec5\u9650\u4e8e\u79d1\u5b66\u7814\u7a76\uff0c\u7edd\u4e0d\u53ef\u4f5c\u4e3a\u52a8\u7269\u6216\u4eba\u7c7b\u75be\u75c5\u7684\u6cbb\u7597\u4ea7\u54c1\u4f7f\u7528\uff08\u4e0e NCI-N87\u3001MKN-45 \u7b49\u7ec6\u80de\u9650\u5236\u6761\u6b3e\u4e00\u81f4\uff0c\u4e25\u7981\u7528\u4e8e\u4e34\u5e8a\u6cbb\u7597\u3001\u7ec6\u80de\u6cbb\u7597\u3001\u7f8e\u5bb9\u53ca\u5176\u4ed6\u975e\u79d1\u7814\u7528\u9014\uff1b\u56e0\u5177\u5907\u4f53\u5185\u4fb5\u88ad\u80fd\u529b\uff0c\u5b9e\u9a8c\u5e9f\u5f03\u7269\u9700\u7ecf 121\u2103\u9ad8\u538b\u706d\u83cc 30 \u5206\u949f\u5904\u7406\uff0c\u907f\u514d\u751f\u7269\u5371\u5bb3\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7279\u522b\u8bf4\u660e\uff1a\u4ee5\u4e0b\u7ec6\u80de\u57f9\u517b\u51bb\u5b58\u5904\u7406\u4ec5\u4f9b\u53c2\u8003\uff0c\u5177\u4f53\u64cd\u4f5c\u6b65\u9aa4\u4ee5\u968f\u8d27\u4ea7\u54c1\u8bf4\u660e\u4e66\u4e3a\u4e3b\uff1b\u8be5\u7ec6\u80de\u5b9e\u9a8c\u64cd\u4f5c\u9700\u91cd\u70b9\u5173\u6ce8\uff1a\u2460\u57f9\u517b\u57fa\u4e0e\u5206\u5316\u7ef4\u62a4\uff1a\u4e25\u683c\u4f7f\u7528 \u201c90% DMEM+10% FBS+PS\u201d \u914d\u65b9\uff0c\u4e0d\u53ef\u4e0e NCI-N87 \u7684 RPMI-1640\u3001MKN-45 \u7684 RPMI-1640 \u6df7\u7528\uff1b\u82e5\u5f00\u5c55\u5206\u5316\u8bf1\u5bfc\u5b9e\u9a8c\uff0c\u53ef\u6dfb\u52a0\u7ef4\u7532\u9178\uff08RA\uff09\uff0c\u901a\u8fc7 HE \u67d3\u8272\u89c2\u5bdf\u817a\u7ba1\u6837\u7ed3\u6784\u53d8\u5316\uff0c\u9a8c\u8bc1\u8bf1\u5bfc\u6548\u679c\uff1b\u2461\u4f53\u5185\u79fb\u690d\u5b9e\u9a8c\u8bbe\u8ba1\uff1a\u6784\u5efa\u76ae\u4e0b\u79fb\u690d\u6a21\u578b\u65f6\uff0c\u9009\u62e9 6-8 \u5468\u9f84\u514d\u75ab\u6291\u5236 ICR \u5c0f\u9f20\uff0c\u63a5\u79cd\u5bc6\u5ea6 1\u00d710\u2076cells \/ \u53ea\uff08\u4f4e\u4e8e NCI-N87 \u7684 1\u00d710\u2077cells \/ \u53ea\uff09\uff0c\u63a5\u79cd\u540e\u6bcf\u5468\u6d4b\u91cf\u80bf\u7624\u4f53\u79ef\uff08\u957f \u00d7 \u5bbd \u00b2\/2\uff09\uff0c3-4 \u5468\u89c2\u5bdf\u808c\u5c42\u4fb5\u88ad\u53ca\u6dcb\u5df4\u7ed3\u8f6c\u79fb\u60c5\u51b5\uff08HE 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