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data-list=\"bullet1\">\u751f\u7269\u5b89\u5168\u7b49\u7ea7\uff1a1\uff08\u4e0e HPAC\u3001CFPAC-1 \u4e00\u81f4\uff0c\u4f4e\u4e8e BEAS-2B \u7684 2 \u7ea7\uff1b\u64cd\u4f5c\u65f6\u9700\u9075\u5faa\u751f\u7269\u5b89\u5168\u4e00\u7ea7\u5b9e\u9a8c\u5ba4\u9632\u62a4\u89c4\u8303\uff0c\u5b9e\u9a8c\u5e9f\u5f03\u7269\u9700\u7ecf 121\u2103\u9ad8\u538b\u706d\u83cc 30 \u5206\u949f\u5904\u7406\uff0c\u907f\u514d\u7ec6\u80de\u6b8b\u7559\u6c61\u67d3\uff1b\u56e0\u57f9\u517b\u6761\u4ef6\u5e38\u89c4\uff0c\u65e0\u9700\u50cf HPAC \u90a3\u6837\u7279\u6b8a\u7ba1\u7406\u6d3b\u6027\u56e0\u5b50\uff0c\u64cd\u4f5c\u6d41\u7a0b\u7b80\u5316\uff0c\u9002\u5408\u65b0\u624b\u64cd\u4f5c\u4eba\u5458\u4f7f\u7528\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 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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\u65e0\u9700\u7279\u6b8a\u63a7\u6e29\u7cbe\u5ea6\uff08\u6ce2\u52a8\u2264\u00b11\u2103\u5373\u53ef\uff0c\u4f4e\u4e8e HPAC \u7684 \u00b10.5\u2103\uff09\uff0c\u65e0\u9700\u50cf HPAC \u90a3\u6837\u62c5\u5fc3\u56e0\u5b50\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 HPAC\u3001CFPAC-1 \u5171\u540c\u653e\u7f6e\uff08\u4ec5\u9700\u533a\u5206\u57f9\u517b\u57fa\uff09\uff0c\u4fbf\u4e8e\u591a\u7ec6\u80de\u682a\u5e76\u884c\u5f00\u5c55 \u201c\u80f0\u817a\u764c\u7ec6\u80de\u591a\u4e9a\u578b\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\u5e38\u89c4\u4e0a\u76ae\u7ec6\u80de\u7279\u6027\uff0c\u65e0\u9700\u7279\u6b8a\u5904\u7406\uff09\uff1a\u2460\u7ec6\u80de\u6536\u96c6\uff1a\u9009\u62e9\u5bf9\u6570\u751f\u957f\u671f\u4e2d\u671f\u7ec6\u80de\uff08\u5bc6\u5ea6 75%\uff0c\u6d3b\u6027\u6700\u4f73\u3001\u5f62\u6001\u7a33\u5b9a\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\u5373\u53ef\u7ec8\u6b62\uff0c\u8f7b\u67d4\u5439\u6253\u907f\u514d\u7ec6\u80de\u7834\u788e\uff08\u65e0\u9700\u50cf HPAC \u90a3\u6837\u7528\u4e13\u7528\u57f9\u517b\u57fa\u8fc7\u6e21\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 HPAC\u3001CFPAC-1 \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 RPMI-1640 \u57f9\u517b\u57fa\u91cd\u60ac\u76f4\u63a5\u63a5\u79cd\uff0c\u907f\u514d DMSO \u5f71\u54cd\u7ec6\u80de\u8d34\u58c1\uff0c\u65e0\u9700\u50cf HPAC \u90a3\u6837\u7acb\u5373\u8865\u5145\u56e0\u5b50\uff0c\u64cd\u4f5c\u6d41\u7a0b\u7b80\u5316\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7ec6\u80de\u8d27\u671f\uff1a1 \u5468\u5de6\u53f3\uff08\u5e93\u5b58\u7ec6\u80de\u5747\u7ecf\u8fc7 RPMI-1640 \u57f9\u517b\u57fa\u9002\u5e94\u6027\u57f9\u517b\u3001\u652f\u539f\u4f53\u68c0\u6d4b\u53ca\u7ec6\u80de\u5f62\u6001\u9a8c\u8bc1\uff08\u4e0a\u76ae\u6837\uff09\uff0c\u6838\u5fc3\u57f9\u517b\u7279\u6027\u7a33\u5b9a\uff0c\u4e0e HPAC\u3001CFPAC-1 \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 HPAC\u3001CFPAC-1 \u540c\u6b65\u4e0b\u5355\uff0c\u65e0\u9700\u8c03\u6574\u65f6\u95f4\uff0c\u4fdd\u969c \u201c\u80f0\u817a\u764c\u7ec6\u80de\u591a\u4e9a\u578b\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\u4e0a\u76ae\u6837\u8d34\u58c1\u7ec6\u80de\u8131\u843d\uff1b\u56e0\u65e0\u9700\u643a\u5e26\u989d\u5916\u56e0\u5b50\uff08\u533a\u522b\u4e8e HPAC\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\u7ec6\u80de\u8d34\u58c1\u72b6\u6001\uff0c48 \u5c0f\u65f6\u5185\u9996\u6b21\u6362\u6db2\uff08\u4f7f\u7528 90% RPMI-1640+10% FBS\uff09\uff0c\u65e0\u9700\u50cf HPAC \u90a3\u6837\u7acb\u5373\u914d\u5236\u4e13\u7528\u57f9\u517b\u57fa\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\uff08\u4f4e\u4e8e HPAC \u7684 1.5kg\uff09\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\u5f62\u6001\u6539\u53d8\u98ce\u9669\uff0c\u56e0\u57f9\u517b\u6761\u4ef6\u5e38\u89c4\uff0c\u8fd0\u8f93\u540e\u7ec6\u80de\u6062\u590d\u901f\u5ea6\u5feb\u4e8e HPAC\u3001CFPAC-1\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 HPAC\u3001CFPAC-1 \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\u5b9e\u9a8c\u8fc7\u7a0b\u4e2d\u9700\u59a5\u5584\u5904\u7406\u542b\u80f0\u817a\u764c\u7ec6\u80de\u7684\u57f9\u517b\u57fa\uff0c\u907f\u514d\u73af\u5883\u66b4\u9732\uff0c\u82e5\u540e\u7eed\u901a\u8fc7\u5b9e\u9a8c\u660e\u786e\u75c5\u7406\u4e9a\u578b\uff0c\u9700\u5728\u5b9e\u9a8c\u8bb0\u5f55\u4e2d\u8865\u5145\u6807\u6ce8\uff0c\u786e\u4fdd\u6570\u636e\u5b8c\u6574\u6027\u53ca\u5b9e\u9a8c\u53ef\u590d\u73b0\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\u2460\u75c5\u7406\u4e9a\u578b\u8865\u5145\u9a8c\u8bc1\uff1a\u9996\u6b21\u57f9\u517b\u65f6\uff0c\u5efa\u8bae\u901a\u8fc7\u514d\u75ab\u8367\u5149\u6216 Western Blot \u68c0\u6d4b\u80f0\u817a\u764c\u4e3b\u8981\u75c5\u7406\u4e9a\u578b\u6807\u5fd7\u7269\uff0c\u660e\u786e\u7ec6\u80de\u5206\u7c7b\uff1aa. \u817a\u6ce1\u764c\u6807\u5fd7\u7269\uff08\u80f0\u86cb\u767d\u9176\u539f\u3001\u8102\u80aa\u9176\uff0c\u4e0e HPAC \u5bf9\u6bd4\uff09\uff1bb. \u5bfc\u7ba1\u764c\u6807\u5fd7\u7269\uff08CA19-9\u3001CK19\uff0c\u4e0e CFPAC-1 \u5bf9\u6bd4\uff09\uff1bc. 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15%-25%\uff08\u53c2\u8003\u540c\u7c7b\u5e38\u89c4\u80f0\u817a\u764c\u7ec6\u80de\uff09\uff0c\u8f6c\u67d3\u540e 48 \u5c0f\u65f6\u68c0\u6d4b\u76ee\u7684\u57fa\u56e0\u8868\u8fbe\uff0c\u53ef\u7528\u4e8e\u57fa\u56e0\u529f\u80fd\u7814\u7a76\u53ca siRNA \u5e72\u6270\u5b9e\u9a8c\uff1bb. \u6d41\u5f0f\u7ec6\u80de\u672f\u5206\u6790\uff1a\u56e0\u7ec6\u80de\u6613\u83b7\u5f97\u5355\u7ec6\u80de\u60ac\u6db2\uff0c\u9002\u5408\u68c0\u6d4b\u7ec6\u80de\u5468\u671f\u3001\u51cb\u4ea1\u7387\uff08\u5982 Annexin V\/PI \u53cc\u67d3\uff09\uff0c\u4e0e HPAC\uff08\u514b\u9686\u751f\u957f\uff09\u3001CFPAC-1\uff08\u6781\u5316\uff09\u76f8\u6bd4\uff0c\u68c0\u6d4b\u7ed3\u679c\u66f4\u51c6\u786e\uff1b\u2463\u957f\u671f\u57f9\u517b\u76d1\u6d4b\uff1a\u6bcf 5 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