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2\uff0c\u4fbf\u4e8e\u5b9e\u9a8c\u5ba4\u7edf\u4e00\u7ba1\u7406\u4e0d\u540c\u5b89\u5168\u7b49\u7ea7\u7684\u7ec6\u80de\u682a\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7ec6\u80de\u89c4\u683c\uff1a1\u00d710\u2076cells\/T25 \u57f9\u517b\u74f6\u6216\u8005 1mL \u51bb\u5b58\u7ba1\u5305\u88c5\uff08\u51bb\u5b58\u7ba1\u542b 10% \u4e8c\u7532\u57fa\u4e9a\u781c\uff08DMSO\uff09\u65e0\u8840\u6e05\u51bb\u5b58\u6db2\uff0c\u7ecf\u68c0\u6d4b\u590d\u82cf\u5b58\u6d3b\u7387\u7a33\u5b9a\u5728 85% \u4ee5\u4e0a\uff0c\u4e14\u9cde\u764c\u5f62\u6001\u3001\u77ed\u500d\u589e\u65f6\u95f4\u7279\u6027\u65e0\u663e\u8457\u6539\u53d8\uff1bT25 \u74f6\u53d1\u8d27\u65f6\u7ec6\u80de\u5bc6\u5ea6\u8fbe 70%-80%\uff0c\u786e\u4fdd\u5230\u8fbe\u540e\u53ef\u76f4\u63a5\u7528\u4e8e\u5b9e\u9a8c\u6216\u5feb\u901f\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\u5feb\u901f\u589e\u6b96\u53ca\u9cde\u764c\u8868\u578b\uff0c\u4fdd\u969c\u5b9e\u9a8c\u5468\u671f\u7a33\u5b9a\u6027\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u4fdd\u85cf\u673a\u6784\uff1a\u4e2d\u56fd\u5178\u578b\u57f9\u517b\u7269\u4fdd\u85cf\u4e2d\u5fc3\u7ec6\u80de\u5e93\uff1bAddexBio; C0013001\/37\uff08\u56fd\u5185\u6743\u5a01\u673a\u6784\u4e0e\u56fd\u9645\u5546\u4e1a\u673a\u6784\u53cc\u91cd\u4fdd\u85cf\uff0c\u7ec6\u80de\u6765\u6e90\u53ef\u8ffd\u6eaf\uff0c\u9cde\u764c\u7279\u6027\u53ca\u751f\u7269\u5b66\u8868\u578b\u7ecf\u8fc7\u4e25\u683c\u9a8c\u8bc1\uff0c\u4e0e WPMY-1\uff08ATCC + \u56fd\u5185\u4fdd\u85cf\uff09\u7684\u4fdd\u85cf\u4f53\u7cfb\u4e92\u8865\uff0c\u4fbf\u4e8e\u56fd\u5185\u5916\u79d1\u7814\u673a\u6784\u534f\u4f5c\u53ca\u6587\u732e\u7ed3\u679c\u590d\u73b0\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u57f9\u517b\u57fa\uff1a90% RPMI-1640+10% FBS+PS\uff08\u9752\u9709\u7d20 &#8211; \u94fe\u9709\u7d20\uff09\uff08\u6838\u5fc3\u7279\u8272\uff1a\u2460\u91c7\u7528 RPMI-1640 \u57f9\u517b\u57fa\uff0c\u533a\u522b\u4e8e WPMY-1 \u7684 DMEM\uff085% FBS\uff09\u3001VCaP \u7684 DMEM\uff0810% FBS\uff09\uff1bRPMI-1640 \u542b\u6709\u7684\u6c28\u57fa\u9178\u7ec4\u6210\u66f4\u9002\u914d\u98df\u7ba1\u9cde\u764c\u7ec6\u80de\u7684\u4ee3\u8c22\u9700\u6c42\uff0c\u652f\u6301\u5176\u5feb\u901f\u589e\u6b96\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\u6742\u86cb\u767d\u5f71\u54cd\u7ec6\u80de\u9cde\u764c\u5206\u5316\u8868\u578b\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\u589e\u6b96\u901f\u5ea6\u53ca\u81f4\u7624\u6027\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\u57f9\u517b\u7bb1\u5185\u6258\u76d8\u9700\u6c34\u5e73\u653e\u7f6e\uff0c\u907f\u514d\u57f9\u517b\u57fa\u503e\u659c\u5bfc\u81f4\u7ec6\u80de\u5206\u5e03\u4e0d\u5747\uff1b\u4e0e WPMY-1\u3001VCaP\u3001PC-3 \u7b49\u6240\u6709\u5df2\u68b3\u7406\u7ec6\u80de\u7684\u57f9\u517b\u6761\u4ef6\u5b8c\u5168\u4e00\u81f4\uff0c\u4ec5\u9700\u9002\u914d\u7ec6\u80de\u81ea\u8eab\u589e\u6b96\u8282\u594f\u8c03\u6574\u4f20\u4ee3\u95f4\u9694\uff0c\u4fbf\u4e8e\u591a\u7ec6\u80de\u682a\u5e76\u884c\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\u77ed\u500d\u589e\u65f6\u95f4\u7279\u6027\uff1a\u2460\u9009\u62e9\u5bf9\u6570\u751f\u957f\u671f\u7ec6\u80de\uff08\u63a5\u79cd\u540e 24-36 \u5c0f\u65f6\uff09\uff0c\u7528 0.25% \u80f0\u9176 &#8211; EDTA \u5b75\u80b2 1.5-2.5 \u5206\u949f\uff08\u77ed\u4e8e WPMY-1 \u7684 3-4 \u5206\u949f\uff09\uff0c\u955c\u4e0b\u89c1\u7ec6\u80de\u95f4\u9699\u589e\u5927\u5373\u53ef\u7ec8\u6b62\uff0c\u8f7b\u67d4\u5439\u6253\u907f\u514d\u7ec6\u80de\u7834\u788e\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 2\u00d710\u2076cells\/mL\uff08\u9ad8\u4e8e\u5e38\u89c4\u7ec6\u80de\uff09\uff0c\u63d0\u5347\u590d\u82cf\u540e\u7ec6\u80de\u5bc6\u5ea6\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 RPMI-1640 \u57f9\u517b\u57fa\u91cd\u60ac\uff0c\u76f4\u63a5\u63a5\u79cd\uff0c\u51cf\u5c11\u5bf9\u7ec6\u80de\u5feb\u901f\u589e\u6b96\u7279\u6027\u7684\u5f71\u54cd\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u500d\u589e\u65f6\u95f4\uff1a~28 hours\uff08\u589e\u6b96\u901f\u5ea6\u5feb\uff0c\u662f\u5df2\u68b3\u7406\u7ec6\u80de\u4e2d\u500d\u589e\u65f6\u95f4\u6700\u77ed\u7684\u7ec6\u80de\u4e4b\u4e00\uff08\u5feb\u4e8e VCaP \u7684 53-144 \u5c0f\u65f6\u3001WPMY-1 \u7684\u5e73\u7f13\u589e\u6b96\uff09\uff0c\u5bf9\u6570\u751f\u957f\u671f\u4e3a\u63a5\u79cd\u540e 12-36 \u5c0f\u65f6\uff0c\u9700\u6bcf\u5929\u955c\u68c0\u7ec6\u80de\u5bc6\u5ea6\uff0c\u5f85\u5bc6\u5ea6\u8fbe 80% \u65f6\u7acb\u5373\u4f20\u4ee3\uff08\u907f\u514d\u8fc7\u5ea6\u5bc6\u96c6\u5bfc\u81f4\u7ec6\u80de\u8001\u5316\uff09\uff1b\u5b9e\u9a8c\u8bbe\u8ba1\u4e2d\u53ef\u5229\u7528\u8be5\u7279\u6027\uff0c\u7f29\u77ed\u836f\u7269\u5904\u7406\u5468\u671f\uff08\u5982 48 \u5c0f\u65f6\uff09\uff0c\u5feb\u901f\u83b7\u53d6\u5b9e\u9a8c\u7ed3\u679c\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u81f4\u7624\u6027\uff1aYes, in nude mice.\uff08\u660e\u786e\u7684\u88f8\u9f20\u6210\u7624\u80fd\u529b\uff0c\u4e14\u9002\u914d BALB\/c \u88f8\u9f20\uff08\u5e38\u7528\u514d\u75ab\u7f3a\u9677\u5c0f\u9f20\u54c1\u7cfb\uff09\uff0c\u6210\u7624\u6548\u7387\u9ad8\u3001\u5468\u671f\u77ed\uff1b\u5efa\u8bae\u63a5\u79cd\u5bc6\u5ea6\u4e3a 1\u00d710\u2076cells \/ \u53ea\uff08\u88f8\u9f20\u76ae\u4e0b\uff09\uff0c\u63a5\u79cd\u540e\u6bcf\u5468\u6d4b\u91cf\u80bf\u7624\u4f53\u79ef\uff08\u957f \u00d7 \u5bbd \u00b2\/2\uff09\uff0c2-3 \u5468\u5373\u53ef\u7ed8\u5236\u5b8c\u6574\u751f\u957f\u66f2\u7ebf\uff0c\u9002\u5408\u5f00\u5c55\u9ad8\u901a\u91cf\u4f53\u5185\u836f\u7269\u7b5b\u9009\u5b9e\u9a8c\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7ec6\u80de\u8d27\u671f\uff1a1 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