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70%-80%\uff1b\u56e0\u6df7\u5408\u5f62\u6001\u7ec6\u80de\u5bf9\u6e29\u5ea6\u654f\u611f\uff0c\u57f9\u517b\u7bb1\u9700\u7a33\u5b9a\u63a7\u6e29\uff08\u6ce2\u52a8\u2264\u00b10.5\u2103\uff09\uff0c\u907f\u514d\u6e29\u5ea6\u53d8\u5316\u5bfc\u81f4\u5f62\u6001\u6bd4\u4f8b\u5931\u8861\uff1b\u57f9\u517b\u73af\u5883\u4e0e\u5176\u4ed6\u80bf\u7624\u7ec6\u80de\u517c\u5bb9\uff0c\u4ec5\u9700\u6ce8\u610f\u57f9\u517b\u57fa\u5dee\u5f02\uff0c\u4fbf\u4e8e\u591a\u7ec6\u80de\u682a\u5e76\u884c\u5f00\u5c55 \u201c\u9aa8\u80bf\u7624 &#8211; \u5185\u810f\u80bf\u7624\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\u6df7\u5408\u5f62\u6001\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%\uff0c\u4e24\u79cd\u5f62\u6001\u6bd4\u4f8b\u6700\u4f73\uff09\uff0c0.25% \u80f0\u9176 &#8211; EDTA \u5b75\u80b2 3-4 \u5206\u949f\uff08\u517c\u987e\u6210\u7ea4\u7ef4\u6837\u7ec6\u80de\u7684\u957f\u6d88\u5316\u65f6\u95f4\uff09\uff0c\u671f\u95f4\u8f7b\u67d4\u62cd\u6253\u57f9\u517b\u74f6\u58c1\u8f85\u52a9\u5206\u6563\uff0c\u955c\u4e0b\u89c1\u4e24\u79cd\u5f62\u6001\u7ec6\u80de\u5747\u8131\u843d\u5373\u53ef\u7ec8\u6b62\uff1b\u2461\u51bb\u5b58\u6d53\u5ea6\uff1a1000rpm \u79bb\u5fc3 5 \u5206\u949f\uff0c\u7528\u9884\u51b7\u65e0\u8840\u6e05\u51bb\u5b58\u6db2\u91cd\u60ac\u81f3 3\u00d710\u2076cells\/mL\uff08\u9ad8\u4e8e\u5e38\u89c4\u7ec6\u80de\u7684 2.5\u00d710\u2076\uff09\uff0c\u8865\u507f\u6df7\u5408\u5f62\u6001\u7ec6\u80de\u7684\u590d\u82cf\u635f\u5931\uff1b\u2462\u51bb\u5b58\u6d41\u7a0b\uff1a4\u2103\u5e73\u8861 30 \u5206\u949f\u2192-20\u2103\u653e\u7f6e 2 \u5c0f\u65f6\u2192-80\u2103\u8fc7\u591c\u2192\u6db2\u6c2e\uff1b\u590d\u82cf\u65f6 37\u2103\u6c34\u6d74 1.5 \u5206\u949f\uff0c\u7acb\u5373\u7528\u9884\u70ed DMEM \u91cd\u60ac\uff0c\u63a5\u79cd\u540e 24 \u5c0f\u65f6\u5185\u4e0d\u6362\u6db2\uff0c\u5e2e\u52a9\u4e24\u79cd\u5f62\u6001\u7ec6\u80de\u540c\u6b65\u6062\u590d\u751f\u957f\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u67d3\u8272\u4f53\uff1a46~50\uff08\u8fd1\u4e8c\u500d\u4f53\uff0c\u67d3\u8272\u4f53\u6570\u76ee\u4ecb\u4e8e BxPC-3 \u7684\u672a\u660e\u786e\u6838\u578b\u4e0e PL45 \u7684\u672a\u660e\u786e\u6838\u578b\u4e4b\u95f4\uff09\uff1a\u2460\u6838\u578b\u7279\u5f81\uff1a\u67d3\u8272\u4f53\u6570\u76ee\u63a5\u8fd1\u6b63\u5e38\u4eba\u4f53\u7ec6\u80de\uff0846 \u6761\uff09\uff0c\u4f53\u73b0\u9752\u5c11\u5e74\u9aa8\u8089\u7624\u7684\u4f4e\u5f02\u500d\u4f53\u7279\u5f81\uff08\u533a\u522b\u4e8e BxPC-3 \u7684\u9ad8\u81f4\u7624\u6027\u6838\u578b\uff09\uff0c\u6838\u578b\u7a33\u5b9a\u6027\u4e2d\u7b49\uff1b\u2461\u5e94\u7528\u573a\u666f\uff1aa. \u6838\u578b\u5206\u6790\uff1a\u901a\u8fc7 G \u5e26\u6280\u672f\u89c2\u5bdf\u67d3\u8272\u4f53\u7ed3\u6784\u5f02\u5e38\uff08\u5982\u4e0e\u9aa8\u8089\u7624\u76f8\u5173\u7684 13 \u53f7\u67d3\u8272\u4f53\u7f3a\u5931\uff09\uff1bb. \u5bf9\u6bd4\u5b9e\u9a8c\uff1a\u4e0e BxPC-3\uff08\u672a\u660e\u786e\uff09\u5bf9\u6bd4\uff0c\u5206\u6790\u9752\u5c11\u5e74\u4e0e\u8001\u5e74\u80bf\u7624\u7684\u6838\u578b\u5dee\u5f02\uff1b\u2462\u6ce8\u610f\u4e8b\u9879\uff1a\u957f\u671f\u57f9\u517b\u9700\u6bcf 5 \u4ee3\u68c0\u6d4b\u67d3\u8272\u4f53\u6570\u76ee\uff0c\u786e\u4fdd\u7279\u5f81\u6027\u6838\u578b\u6bd4\u4f8b\u7a33\u5b9a\uff0846~50 \u6761\u7ec6\u80de\u5360\u6bd4\u226580%\uff09\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\u6df7\u5408\u5f62\u6001\u9a8c\u8bc1\uff08\u4e0a\u76ae\u6837\uff1a\u6210\u7ea4\u7ef4\u6837\u22481:1\uff09\u53ca\u652f\u539f\u4f53\u68c0\u6d4b\uff0c\u6838\u5fc3\u7279\u6027\u7a33\u5b9a\uff1b\u56e0\u57f9\u517b\u6761\u4ef6\u5e38\u89c4\uff0c\u4e0b\u5355\u540e 48 \u5c0f\u65f6\u5185\u53ef\u53d1\u8d27\uff0c\u53ef\u4e0e BxPC-3 \u540c\u6b65\u4e0b\u5355\uff0c\u901a\u8fc7\u8c03\u6574\u63a5\u79cd\u65f6\u95f4\uff08HOS \u665a\u63a5\u79cd 1 \u5929\uff0c\u56e0\u589e\u6b96\u7565\u5feb\uff09\uff0c\u5b9e\u73b0 \u201c\u9aa8\u80bf\u7624 &#8211; \u80f0\u817a\u764c\u201d \u5e73\u884c\u5b9e\u9a8c\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\u53cc\u5c42\u9632\u9707\u6ce1\u6cab\u7bb1\uff0c\u5185\u7f6e 2-8\u2103\u51b0\u888b\uff0c\u8fd0\u8f93\u65f6\u7528\u56fa\u5b9a\u652f\u67b6\u9632\u6b62\u57f9\u517b\u74f6\u6643\u52a8\uff0c\u51cf\u5c11\u6df7\u5408\u5f62\u6001\u7ec6\u80de\u8131\u843d\uff1b\u5230\u8fbe\u540e\u7acb\u5373\u653e\u5165 CO\u2082\u57f9\u517b\u7bb1\uff0c24 \u5c0f\u65f6\u540e\u89c2\u5bdf\u5f62\u6001\u6bd4\u4f8b\uff0c48 \u5c0f\u65f6\u5185\u9996\u6b21\u6362\u6db2\uff08\u4fdd\u7559 50% \u65e7\u57f9\u517b\u57fa\uff09\uff0c\u5e2e\u52a9\u7ec6\u80de\u9002\u5e94\u65b0\u73af\u5883\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%\uff09 \u987a\u4e30\u6b21\u6668\u8fbe\uff0c\u7f29\u77ed\u8fd0\u8f93\u65f6\u95f4\u51cf\u5c11\u5f62\u6001\u6bd4\u4f8b\u6ce2\u52a8<\/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 BxPC-3\u3001PL45 \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\u9752\u5c11\u5e74\u6765\u6e90\uff0c\u989d\u5916\u9650\u5236\uff1a\u2460\u4e0d\u53ef\u7528\u4e8e\u9752\u5c11\u5e74\u76f8\u5173\u7684\u4eba\u4f53\u5b9e\u9a8c\uff1b\u2461\u5b9e\u9a8c\u6570\u636e\u53d1\u8868\u9700\u6807\u6ce8\u5e74\u9f84\u3001\u6027\u522b\u53ca\u79cd\u65cf\u80cc\u666f\uff0c\u786e\u4fdd\u7814\u7a76\u4e25\u8c28\u6027\uff09<\/li>\n<\/ul>\n<ul>\n<li class=\"temp-li bullet1\" data-line=\"true\" data-list=\"bullet1\">\u7279\u522b\u8bf4\u660e\uff1a\u6838\u5fc3\u5efa\u8bae\uff1a\u2460\u6df7\u5408\u5f62\u6001\u7ec6\u80de\u64cd\u4f5c\uff1aa. \u4f20\u4ee3\u6280\u5de7\uff1a\u6d88\u5316\u65f6\u5148\u89c2\u5bdf\u6210\u7ea4\u7ef4\u6837\u7ec6\u80de\uff08\u68ad\u5f62\uff09\u662f\u5426\u8131\u843d\uff0c\u5f85\u5176\u8131\u843d 50% \u65f6\uff0c\u8f7b\u8f7b\u5439\u6253\u4e0a\u76ae\u6837\u7ec6\u80de\u533a\u57df\uff0c\u907f\u514d\u8fc7\u5ea6\u6d88\u5316\u5bfc\u81f4\u4e0a\u76ae\u6837\u7ec6\u80de\u635f\u4f24\uff1bb. \u5f62\u6001\u7ef4\u6301\uff1a\u4f20\u4ee3\u65f6\u786e\u4fdd\u4e24\u79cd\u5f62\u6001\u7ec6\u80de\u5747\u5300\u63a5\u79cd\uff0c\u907f\u514d\u5355\u4e00\u5f62\u6001\u7ec6\u80de\u5bcc\u96c6\uff08\u5982\u6210\u7ea4\u7ef4\u6837\u7ec6\u80de\u8fc7\u5ea6\u751f\u957f\uff09\uff1b\u82e5\u5f62\u6001\u6bd4\u4f8b\u5931\u8861\uff08\u67d0\u4e00\u5f62\u6001\u5360\u6bd4\uff1c30%\uff09\uff0c\u9700\u91cd\u65b0\u590d\u82cf\u4f4e\u4ee3\u6570\u7ec6\u80de\uff1b\u2461\u9aa8\u8089\u7624\u7279\u6027\u9a8c\u8bc1\uff1a\u9996\u6b21\u57f9\u517b\u65f6\uff0c\u5efa\u8bae\u901a\u8fc7\u4ee5\u4e0b\u5b9e\u9a8c\u786e\u8ba4\u9aa8\u8089\u7624\u5c5e\u6027\uff1aa. \u78b1\u6027\u78f7\u9178\u9176\u67d3\u8272\uff08\u9aa8\u8089\u7624\u7ec6\u80de\u9633\u6027\uff0cBxPC-3 \u9634\u6027\uff09\uff1bb. \u9aa8\u6865\u86cb\u767d Western Blot \u68c0\u6d4b\uff08\u9ad8\u8868\u8fbe\uff0c\u533a\u522b\u4e8e\u80f0\u817a\u764c\u7684\u4f4e\u8868\u8fbe\uff09\uff1bc. 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