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bullet1\" data-line=\"true\" data-list=\"bullet1\">\u57f9\u517b\u57fa\uff1a90%1640\uff08RPMI-1640\uff09+10% FBS+PS\uff08\u9752\u9709\u7d20 &#8211; \u94fe\u9709\u7d20\uff09\uff08\u6838\u5fc3\u7279\u8272\uff0c\u4e0e MG-63 \u5bf9\u6bd4\uff09\uff1a\u2460\u5e38\u89c4\u914d\u65b9\u5dee\u5f02\uff1a\u91c7\u7528 RPMI-1640 \u57f9\u517b\u57fa\uff0c\u533a\u522b\u4e8e MG-63 \u7684 MEM (\u542b NEAA)\u3001HOS \u7684 DMEM\uff1b1640 \u542b\u9002\u914d\u7ea4\u7ef4\u539f\u7ec6\u80de\u6837\u7ec6\u80de\u7684\u8425\u517b\u914d\u6bd4\uff08\u5982\u9002\u91cf\u6c28\u57fa\u9178\u3001\u7ef4\u751f\u7d20\uff09\uff0c\u652f\u6301\u539f\u53d1\u6027\u9aa8\u8089\u7624\u7ec6\u80de\u7684\u57fa\u7840\u589e\u6b96\u53ca p53 \u76f8\u5173\u57fa\u56e0\u529f\u80fd\u7ef4\u6301\uff0c10% FBS \u4e3a\u5e38\u89c4\u8840\u6e05\u6d53\u5ea6\uff08\u4e0e MG-63\u3001HOS \u4e00\u81f4\uff09\uff0c\u65e0\u7279\u6b8a\u6dfb\u52a0\u56e0\u5b50\uff08\u533a\u522b\u4e8e MG-63 \u7684\u65e0\u989d\u5916\u56e0\u5b50\u3001PANC 05.04 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\u7a7a\u6c14 + 5% \u4e8c\u6c27\u5316\u78b3\uff1b\u6e29\u5ea6\uff1a37\u2103\uff08\u4e0e MG-63\u3001HOS \u4e00\u81f4\uff0c\u6e7f\u5ea6\u7ef4\u6301\u5728 70%-80%\uff0c\u5b9a\u671f\u66f4\u6362\u6c34\u76d8\u9632\u6b62\u57f9\u517b\u57fa\u84b8\u53d1\uff1b\u57f9\u517b\u7bb1\u65e0\u9700\u7279\u6b8a\u63a7\u6e29\u7cbe\u5ea6\uff08\u6ce2\u52a8\u2264\u00b11\u2103\u5373\u53ef\uff0c\u4f4e\u4e8e MG-63 \u7684 \u00b10.5\u2103\uff09\uff0c\u65e0\u9700\u50cf MG-63 \u90a3\u6837\u62c5\u5fc3\u8bf1\u5bfc\u5242\u6d3b\u6027\u53d7\u6e29\u5ea6\u5f71\u54cd\uff1b\u57f9\u517b\u73af\u5883\u4e0e\u5176\u4ed6\u80bf\u7624\u7ec6\u80de\u517c\u5bb9\u6027\u9ad8\uff0c\u53ef\u4e0e MG-63\u3001HOS \u5171\u540c\u653e\u7f6e\uff08\u4ec5\u9700\u533a\u5206\u57f9\u517b\u57fa\uff09\uff0c\u4fbf\u4e8e\u591a\u9aa8\u8089\u7624\u7ec6\u80de\u7cfb\u5e76\u884c\u5f00\u5c55 \u201c\u90e8\u4f4d &#8211; \u4e9a\u578b &#8211; \u57fa\u56e0\u201d \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\u7ea4\u7ef4\u539f\u7ec6\u80de\u6837\u7279\u6027\uff09\uff1a\u2460\u7ec6\u80de\u6536\u96c6\uff1a\u9009\u62e9\u5bf9\u6570\u751f\u957f\u671f\u4e2d\u671f\u7ec6\u80de\uff08\u5bc6\u5ea6 75%\uff0cp53 \u76f8\u5173\u57fa\u56e0\u8868\u8fbe\u6700\u4f73\uff09\uff0c\u7528 0.25% \u80f0\u9176 &#8211; EDTA \u5b75\u80b2 3-4 \u5206\u949f\uff0c\u955c\u4e0b\u89c1\u7ec6\u80de\u95f4\u9699\u589e\u5927\u5373\u53ef\u7ec8\u6b62\uff0c\u8f7b\u67d4\u5439\u6253\u907f\u514d\u7ec6\u80de\u635f\u4f24\uff08\u65e0\u9700\u50cf MG-63 \u90a3\u6837\u8003\u8651\u53d7\u4f53\u6d3b\u6027\uff09\uff1b\u2461\u51bb\u5b58\u6d53\u5ea6\uff1a1000rpm \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 2.5\u00d710\u2076cells\/mL\uff08\u4e0e MG-63\u3001HOS \u4e00\u81f4\uff09\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 1640 \u57f9\u517b\u57fa\u91cd\u60ac\u76f4\u63a5\u63a5\u79cd\uff0c\u907f\u514d DMSO \u5f71\u54cd\u7ec6\u80de\u8d34\u58c1\u53ca p53 \u76f8\u5173\u57fa\u56e0\u529f\u80fd\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 1640 \u57f9\u517b\u57fa\u9002\u5e94\u6027\u57f9\u517b\u3001\u652f\u539f\u4f53\u68c0\u6d4b\u53ca\u7ec6\u80de\u5f62\u6001\u9a8c\u8bc1\uff08\u7ea4\u7ef4\u539f\u7ec6\u80de\u6837\uff09\uff0c\u6838\u5fc3\u57f9\u517b\u7279\u6027\u7a33\u5b9a\uff0c\u4e0e MG-63\u3001HOS \u7684 1 \u5468\u8d27\u671f\u4e00\u81f4\uff1b\u56e0\u57f9\u517b\u57fa\u5e38\u89c4\u4e14\u65e0\u9700\u7279\u6b8a\u9884\u5904\u7406\uff0c\u4e0b\u5355\u540e 48 \u5c0f\u65f6\u5185\u53ef\u5b89\u6392\u53d1\u8d27\uff0c\u53ef\u4e0e MG-63\u3001HOS \u540c\u6b65\u4e0b\u5355\uff0c\u65e0\u9700\u8c03\u6574\u65f6\u95f4\uff0c\u4fdd\u969c \u201c\u591a\u9aa8\u8089\u7624\u7ec6\u80de\u7cfb\u201d \u5e73\u884c\u5b9e\u9a8c\u7684\u65f6\u95f4\u4e00\u81f4\u6027\uff0c\u663e\u8457\u63d0\u5347\u5b9e\u9a8c\u6548\u7387\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\u7528\u56fa\u5b9a\u652f\u67b6\u9632\u6b62\u57f9\u517b\u74f6\u6643\u52a8\uff0c\u51cf\u5c11\u7ea4\u7ef4\u539f\u7ec6\u80de\u6837\u8d34\u58c1\u7ec6\u80de\u8131\u843d\uff1b\u56e0\u65e0\u9700\u643a\u5e26\u989d\u5916\u8bd5\u5242\uff08\u533a\u522b\u4e8e MG-63 \u7684 NEAA\uff09\uff0c\u5305\u88c5\u66f4\u7b80\u6d01\uff0c\u8fd0\u8f93\u98ce\u9669\u964d\u4f4e\uff1b\u5230\u8fbe\u540e\u7acb\u5373\u653e\u5165 5% CO\u2082\u57f9\u517b\u7bb1\uff0c24 \u5c0f\u65f6\u540e\u89c2\u5bdf\u8d34\u58c1\u72b6\u6001\uff0c48 \u5c0f\u65f6\u5185\u9996\u6b21\u6362\u6db2\uff08\u4f7f\u7528 90%1640+10% FBS+PS\uff09\uff0c\u7ec6\u80de\u9002\u5e94\u73af\u5883\u66f4\u5feb\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\u914d\u5907 1kg \u5e72\u51b0\u5373\u53ef\uff0c\u786e\u4fdd\u5230\u8fbe\u65f6\u5e72\u51b0\u5269\u4f59\u91cf\u226560%\uff0c\u7ef4\u6301\u7ec6\u80de\u51bb\u5b58\u72b6\u6001\uff09 \u987a\u4e30\u5feb\u9012\uff08\u4f18\u5148\u9009\u62e9\u6b21\u6668\u8fbe\u670d\u52a1\uff0c\u6700\u5927\u9650\u5ea6\u7f29\u77ed\u8fd0\u8f93\u65f6\u95f4\uff0c\u964d\u4f4e\u7ec6\u80de\u6d3b\u6027\u635f\u5931\u53ca p53 \u76f8\u5173\u57fa\u56e0\u8868\u8fbe\u6ce2\u52a8\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 MG-63\u3001HOS \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\u542b p53 \u76f8\u5173\u57fa\u56e0\uff0c\u989d\u5916\u9650\u5236\uff1a\u2460\u4e0d\u53ef\u7528\u4e8e p53 \u76f8\u5173\u7684\u4eba\u4f53\u57fa\u56e0\u6cbb\u7597\u5b9e\u9a8c\uff1b\u2461\u5b9e\u9a8c\u8fc7\u7a0b\u4e2d\u9700\u59a5\u5584\u5904\u7406\u542b\u8be5\u7ec6\u80de\u7684\u57f9\u517b\u57fa\uff0c\u907f\u514d\u73af\u5883\u66b4\u9732\uff1b\u2462\u82e5\u540e\u7eed\u660e\u786e\u6027\u522b\u5e74\u9f84\u6216 p53 \u57fa\u56e0\u5177\u4f53\u7c7b\u578b\uff0c\u9700\u5728\u5b9e\u9a8c\u8bb0\u5f55\u4e2d\u8865\u5145\u6807\u6ce8\uff0c\u786e\u4fdd\u6570\u636e\u5b8c\u6574\u6027\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\u2460p53 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MG-63\uff08\u6210\u9aa8\u8089\u7624 \/ TGF-\u03b2+\uff09\u3001HOS\uff08\u9aa8\u8089\u7624 \/ \u6df7\u5408\u5f62\u6001\uff09\u5f00\u5c55\u5b9e\u9a8c\u65f6\uff0c\u53ef\uff1aa. \u68c0\u6d4b\u539f\u53d1\u6027\u9aa8\u8089\u7624\u6807\u5fd7\u7269\uff08\u5982 OPN\u3001COL1A1\uff0cSJSA-1 \u9ad8\u8868\u8fbe\uff09\uff0c\u9a8c\u8bc1\u90e8\u4f4d\u7279\u5f02\u6027\uff1bb. \u5bf9\u6bd4 p53 \u901a\u8def\u6d3b\u6027\uff08\u5982 DNA \u635f\u4f24\u540e\u51cb\u4ea1\u7387\uff0cSJSA-1 \u56e0\u542b p53 \u76f8\u5173\u57fa\u56e0\u53ef\u80fd\u66f4\u9ad8\uff09\uff1bc. \u5206\u6790\u836f\u7269\u654f\u611f\u6027\uff08\u5982\u5bf9 p53 \u6fc0\u6d3b\u5242\u7684\u54cd\u5e94\uff0cSJSA-1 \u53ef\u80fd\u66f4\u654f\u611f\uff09\uff1b\u2462\u57fa\u7840\u5b9e\u9a8c\u5e94\u7528\uff1a\u56e0\u5e38\u89c4\u57f9\u517b\u6761\u4ef6\u53ca\u7ea4\u7ef4\u539f\u7ec6\u80de\u6837\u5f62\u6001\uff0c\u9002\u5408\u5f00\u5c55\uff1aa. \u57fa\u56e0\u529f\u80fd\u5b9e\u9a8c\uff1a\u901a\u8fc7 siRNA \u5e72\u6270 p53 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