{"id":7253,"date":"2024-06-06T16:46:06","date_gmt":"2024-06-06T08:46:06","guid":{"rendered":"https:\/\/chimebiologics.com\/?p=7253"},"modified":"2024-12-12T13:28:20","modified_gmt":"2024-12-12T05:28:20","slug":"cho-cell-line-development-process-and-key-technical-points-overview","status":"publish","type":"post","link":"https:\/\/chimebiologics.com\/zh\/cho-cell-line-development-process-and-key-technical-points-overview\/","title":{"rendered":"CHO\u7ec6\u80de\u682a\u5f00\u53d1\uff1a\u6d41\u7a0b\u4e0e\u5173\u952e\u6280\u672f\u70b9\u6982\u8ff0"},"content":{"rendered":"<style type=\"text\/css\" data-created_by=\"avia_inline_auto\" id=\"style-css-av-iwqe2-97bb6aa3783a5420eb8a707e0ad897c8\">\n.flex_column.av-iwqe2-97bb6aa3783a5420eb8a707e0ad897c8{\n-webkit-border-radius:0px 0px 0px 0px;\n-moz-border-radius:0px 0px 0px 0px;\nborder-radius:0px 0px 0px 0px;\npadding:0px 0px 0px 0px;\n}\n<\/style>\n<div class='flex_column av-iwqe2-97bb6aa3783a5420eb8a707e0ad897c8 av_two_third  avia-builder-el-0  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https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/05\/3z-1-300x128.png 300w, https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/05\/3z-1-768x327.png 768w, https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/05\/3z-1-705x300.png 705w\" sizes=\"auto, (max-width: 900px) 100vw, 900px\" \/><\/p>\n<p>\u751f\u7269\u836f\u548c\u8bca\u65ad\u8bd5\u5242\u4f5c\u4e3a\u5236\u836f\u5e02\u573a\u4e0a\u589e\u957f\u6700\u5feb\u7684\u4ea7\u54c1\uff0c\u53d7\u5230\u4e86\u4eba\u4eec\u7684\u5173\u6ce8\u548c\u5e7f\u6cdb\u7684\u5e94\u7528\u3002\u5176\u4e2d\u5305\u62ec\u4e86\u5404\u79cd\u5355\u514b\u9686\u6297\u4f53\uff08mAb\uff09\u3001\u91cd\u7ec4\u86cb\u767d\u75ab\u82d7\u3001\u6fc0\u7d20\u548c\u5176\u4ed6\u86cb\u767d\u8d28\u7c7b\u4ea7\u54c1\uff0c\u968f\u7740\u8fd9\u4e9b\u751f\u7269\u5236\u5242\u8d8a\u6765\u8d8a\u591a\u5730\u8fdb\u5165\u5e02\u573a\uff0c\u8fd9\u4e2a\u6570\u5b57\u8fd8\u5728\u4e0d\u65ad\u589e\u957f\u3002\u8fd1\u51e0\u5e74\uff0c\u5e73\u5747\u6bcf\u5e74\u670915 \u79cd\u4ee5\u4e0a\u91cd\u7ec4\u86cb\u767d\u65b0\u836f\u901a\u8fc7\u7f8e\u56fd\u98df\u54c1\u836f\u54c1\u76d1\u7763\u7ba1\u7406\u5c40\uff08FDA\uff09\u6279\u51c6\uff0c\u5e76\u4e14\u5728\u51e0\u4e2a\u91cd\u78c5\u751f\u7269\u6297\u4f53\u836f\u4e13\u5229\u5373\u5c06\u5230\u671f\u4fc3\u4f7f\u751f\u7269\u4eff\u5236\u836f\u51fa\u73b0\u7684\u523a\u6fc0\u4e0b\uff0c\u5168\u7403\u751f\u7269\u6297\u4f53\u836f\u9500\u552e\u989d\u5df2\u8d85\u8fc71800\u4ebf\u7f8e\u5143\u6bcf\u5e74\u3002<\/p>\n<p><strong>01 <\/strong><strong>CHO\u7ec6\u80de\u7684\u4f18\u52bf<\/strong><\/p>\n<p>\u54fa\u4e73\u52a8\u7269\u7ec6\u80de\u662f\u4e34\u5e8a\u6297\u4f53\u836f\u7269\u4ea7\u54c1\u4f7f\u7528\u7684\u4e3b\u8981\u8868\u8fbe\u7cfb\u7edf\u3002\u5bf9\u4e8e\u6cbb\u7597\u6027\u6297\u4f53\u800c\u8a00\uff0c\u4e3a\u4e86\u6ee1\u8db3\u5176\u751f\u7269\u6d3b\u6027\uff0c\u9700\u8981\u8fdb\u884c\u6b63\u786e\u7684\u6298\u53e0\u548c\u7ffb\u8bd1\u540e\u4fee\u9970\uff0c\u56e0\u6b64\u7528\u4e8e\u751f\u4ea7\u6cbb\u7597\u6027\u6297\u4f53\u7684\u5bbf\u4e3b\u7ec6\u80de\u5f80\u5f80\u662f\u54fa\u4e73\u52a8\u7269\u7ec6\u80de\uff0c\u4e3b\u8981\u5305\u62ec\uff1aSp2\/0 \u9aa8\u9ad3\u7624\u7ec6\u80de\u3001NS0 \u5c0f\u9f20\u9aa8\u9ad3\u7624\u7ec6\u80de\u3001HEK293\u4eba\u80da\u80ce\u80be\u7ec6\u80de\u548c\u4e2d\u56fd\u4ed3\u9f20\u5375\u5de2\u7ec6\u80de\uff08Chinese hamster ovary\uff0cCHO\uff09\uff0c\u5176\u4e2d\u4ee5CHO\u7ec6\u80de\u7528\u9014\u6700\u4e3a\u5e7f\u6cdb\u3002<\/p>\n<p>\u4e0e\u5176\u4ed6\u8868\u8fbe\u7cfb\u7edf\u76f8\u6bd4, CHO\u8868\u8fbe\u7cfb\u7edf\u5360\u6709\u5f88\u5927\u4f18\u52bf\uff1a<\/p>\n<ul>\n<li>\u66f4\u9002\u7528\u4e8e\u60ac\u6d6e\u57f9\u517b,\u53ef\u4ee5\u6ee1\u8db3\u5927\u89c4\u6a21\u5de5\u4e1a\u751f\u4ea7\u91cd\u7ec4\u86cb\u767d\u7684\u8981\u6c42<\/li>\n<li>\u4ea7\u751f\u7684\u6297\u4f53\u5206\u5b50\u5728\u7ed3\u6784\u3001\u529f\u80fd\u65b9\u9762\u548c\u5929\u7136\u6297\u4f53\u5206\u5b50\u8f83\u63a5\u8fd1<\/li>\n<li>\u6240\u542b\u4eba\u7c7b\u75c5\u6bd2\u6781\u5c11<\/li>\n<li>\u5916\u6e90\u57fa\u56e0\u53ef\u4ee5\u5728CHO\u7ec6\u80de\u4e2d\u7a33\u5b9a\u5730\u6574\u5408<\/li>\n<li>CHO\u7ec6\u80de\u662f\u6210\u7ea4\u7ef4\u7ec6\u80de, \u51e0\u4e4e\u4e0d\u5206\u6ccc\u5185\u6e90\u6027\u86cb\u767d, \u56e0\u6b64\u5bf9\u76ee\u6807\u91cd\u7ec4\u6297\u4f53\u7684\u5206\u79bb\u7eaf\u5316\u5de5\u4f5c\u5341\u5206\u6709\u5229<\/li>\n<\/ul>\n<section>\n<section>\n<section>\n<section>\n<section>\n<section>\n<section>\u73b0\u4eca\uff0c\u5404\u4e2a\u5236\u836f\u516c\u53f8\u503e\u5411\u4e8e\u4f7f\u7528\u76f8\u540c\u7684\u5bbf\u4e3b\u7ec6\u80de\u682a\u8fdb\u884c\u751f\u4ea7\u548c\u521d\u671f\u7684\u4e34\u5e8a\u8bd5\u9a8c\uff0c\u56e0\u4e3a\u5404\u4e2a\u4f01\u4e1a\u610f\u8bc6\u5230\u8fd9\u79cd\u7b56\u7565\u7b80\u5316\u4e86\u7ec6\u80de\u682a\u5f00\u53d1\u5468\u671f\u5e76\u5927\u5927\u52a0\u5feb\u548c\u4f18\u5316\u4e86\u521d\u671f\u4e34\u5e8a\u8bd5\u9a8c\u7684\u65f6\u95f4\u548c\u9014\u5f84\u3002\u6b64\u5916\uff0c\u8fd9\u79cd\u65b9\u6cd5\u8fd8\u964d\u4f4e\u4e86\u4ea7\u54c1\u53ef\u6bd4\u6027\u95ee\u9898\u7684\u98ce\u9669\uff0c\u5e2e\u52a9\u7533\u62a5\u4f01\u4e1a\u6ee1\u8db3\u4ea7\u54c1\u8d28\u91cf\u548c\u836f\u7269\u4e00\u81f4\u6027\u7684\u5e0c\u671b\u3002\u8fd9\u4e9b\u4e0d\u65ad\u53d8\u5316\u548c\u4ee5\u65f6\u95f4\u6548\u7387\u4f18\u5148\u7684\u9879\u76ee\u4e3aCHO\u8868\u8fbe\u5e73\u53f0\u7684\u521b\u65b0\u63d0\u4f9b\u4e86\u65b0\u7684\u6280\u672f\u7814\u7a76\u65b9\u5411\u3002\u5de5\u4e1a\u7ec6\u80de\u682a\u5f00\u53d1\u7684\u91cd\u70b9\u5df2\u8f6c\u5411\u5bbf\u4e3b\u7ec6\u80de\u682a\u7a33\u5b9a\u6027\u95ee\u9898\uff0c\u4ee5\u786e\u4fdd\u957f\u671f\u751f\u4ea7\u7684\u7a33\u5b9a\u6027\uff0c\u4f7f\u7528\u9776\u5411\u6574\u5408\u6280\u672f\u8fdb\u884c\u8868\u8fbe\u4f18\u5316\uff0c\u5c24\u5176\u662f\u590d\u6742\u7684\u91cd\u7ec4\u86cb\u767d\uff0c\u4ee5\u53ca\u5f00\u53d1\u3001\u9009\u62e9\u548c\u7b5b\u9009\u8868\u8fbe\u7cfb\u7edf\u4ee5\u5b9e\u73b0\u66f4\u5feb\u3001\u66f4\u6709\u6548\u7684\u514b\u9686\u9009\u62e9\u3002<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<\/section>\n<section>\n<section><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-7254 aligncenter\" src=\"https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/CHO\u7ec6\u80de\u682a\u5f00\u53d1\u6d41\u7a0b\u56fe.png\" alt=\"\" width=\"525\" height=\"444\" srcset=\"https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/CHO\u7ec6\u80de\u682a\u5f00\u53d1\u6d41\u7a0b\u56fe.png 767w, https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/CHO\u7ec6\u80de\u682a\u5f00\u53d1\u6d41\u7a0b\u56fe-300x253.png 300w, https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/CHO\u7ec6\u80de\u682a\u5f00\u53d1\u6d41\u7a0b\u56fe-705x596.png 705w\" sizes=\"auto, (max-width: 525px) 100vw, 525px\" \/><\/p>\n<p style=\"text-align: center;\"><span style=\"color: #999999;\">CHO\u7ec6\u80de\u682a\u5f00\u53d1\u6d41\u7a0b\u56fe<\/span><\/p>\n<\/section>\n<\/section>\n<p><strong>02\u00a0 CHO\u7ec6\u80de\u682a\u5f00\u53d1\u8fc7\u7a0b\u4e2d\u7684\u5173\u952e\u6280\u672f\u70b9<\/strong><\/p>\n<ul>\n<li>\u8868\u8fbe\u8f7d\u4f53\u7684\u6784\u5efa\u4e0e\u8f6c\u67d3<\/li>\n<\/ul>\n<p>\u7ec6\u80de\u682a\u5f00\u53d1\u9636\u6bb5\u662f\u4ece\u9ad8\u8868\u8fbe\u7684\u57fa\u56e0\u4f18\u5316\u5f00\u59cb\uff0c\u7136\u540e\u5c06\u76ee\u7684\u57fa\u56e0\u63d2\u5165CHO\u5bbf\u4e3b\u7ec6\u80de\u4e2d\u3002\u5e38\u89c1\u7684\u8f6c\u67d3\u65b9\u5f0f\u4e3b\u8981\u6709\u78f7\u9178\u9499\u8f6c\u67d3\u3001\u7535\u7a7f\u5b54\u8f6c\u67d3\u3001\u8102\u8d28\u4f53\u8f6c\u67d3\u548c\u9006\u8f6c\u5f55\u75c5\u6bd2\u8f6c\u67d3\u3002DNA\u8fdb\u5165\u5bbf\u4e3b\u7ec6\u80de\u6838\u540e\u5c06\u4f1a\u968f\u673a\u6574\u5408\u5230\u5bbf\u4e3b\u7ec6\u80de\u57fa\u56e0\u7ec4\u4e2d\uff0c\u56e0\u6b64\u5176\u8868\u8fbe\u6c34\u5e73\u4e0e\u57fa\u56e0\u62f7\u8d1d\u6570\u3001\u57fa\u56e0\u6574\u5408\u4f4d\u70b9\u7684\u8f6c\u5f55\u6d3b\u6027\u6709\u5173\u3002\u88ab\u7b5b\u9009\u51fa\u6765\u7684\u514b\u9686\u7ecf\u8fc7\u4f20\u4ee3\u57f9\u517b\u548c\u5206\u6279\u8865\u6599\uff08fed-batch\uff09\u5b9e\u9a8c\uff0c\u6bd4\u8f83\u5176\u751f\u957f\u7279\u6027\u3001\u4ee3\u8c22\u72b6\u51b5\u3001\u8868\u8fbe\u91cf\u9ad8\u4f4e\u3001\u8868\u8fbe\u4ea7\u7269\u8d28\u91cf\u7b49\u56e0\u7d20\uff0c\u9009\u51fa\u6700\u4f18\u7684\u51e0\u4e2a\u514b\u9686\u8fdb\u5165\u751f\u7269\u53cd\u5e94\u5668\u653e\u5927\u5b9e\u9a8c\uff0c\u6700\u7ec8\u786e\u5b9a\u751f\u4ea7\u7528\u5355\u514b\u9686\u7ec6\u80de\u682a\u3002<br \/>\n\u9002\u5408\u4e8eCHO\u7ec6\u80de\u8868\u8fbe\u7684\u8f7d\u4f53(Vector)\u7684\u57fa\u672c\u5143\u7d20\u5305\u62ec: \u542f\u52a8\u5b50(promoter)\u3001poly A\u52a0\u5c3e\u4fe1\u53f7(poly A signal)\u3001\u7b5b\u9009\u6807\u8bb0( selectable marker)\u3001\u514b\u9686\u4f4d\u70b9(cloning site)\u548c\u590d\u5236\u8d77\u59cb\u4f4d\u70b9(origin of replication)\uff0c\u6709\u4e9b\u8fd8\u6709\u62a5\u544a\u57fa\u56e0( reporter gene)\u3002\u542f\u52a8\u5b50\u4e00\u822c\u8f83\u591a\u91c7\u7528SV40\u548cCMV\u4e24\u79cd\uff0cCMV\u542f\u52a8\u5b50\u901a\u5e38\u4f4d\u4e8e\u76ee\u7684\u57fa\u56e0(\u8f7b\u94fe\u548c\u91cd\u94fe\u53ef\u53d8\u533a\u57fa\u56e0)\u4e4b\u524d\uff0c\u800cSV40\u5219\u4f4d\u4e8e\u7b5b\u9009\u6807\u8bb0\u4e4b\u524d\u3002\u7b5b\u9009\u6807\u8bb0\u4e00\u822c\u6709\u4e24\u79cd\uff1a\u4ee3\u8c22\u578b\u548c\u6297\u751f\u7d20\u578b\uff0c\u6216\u8005\u4e5f\u53ef\u79f0\u4e3a\u6269\u589e\u578b\u57fa\u56e0\u7b5b\u9009\u6807\u8bb0\u548c\u975e\u6269\u589e\u578b\u57fa\u56e0\u7b5b\u9009\u6807\u8bb0\u3002\u5e38\u89c1\u7684\u7b5b\u9009\u6807\u8bb0\u89c1\u4e0b\u8868\uff0c\u5f53\u524d\u5de5\u4e1a\u754c\u8f83\u591a\u91c7\u7528\u7684\u7b5b\u9009\u6807\u8bb0\u4e3aDHFR\u3001GS\u3001G418\u548cpuromycin\u7b49\u3002<\/p>\n<table width=\"100%\">\n<tbody>\n<tr>\n<td colspan=\"1\" rowspan=\"1\" width=\"30.0000%\">\n<section>\n<section><strong>\u7c7b\u522b<\/strong><\/section>\n<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"35.0000%\">\n<section>\n<section><strong>\u7b5b\u9009\u6807\u8bb0<\/strong><\/section>\n<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"34.9900%\">\n<section>\n<section><strong>\u7b5b\u9009\u8bd5\u5242<\/strong><\/section>\n<\/section>\n<\/td>\n<\/tr>\n<tr>\n<td colspan=\"1\" rowspan=\"2\" width=\"30.0000%\">\n<section>\n<section>\u4ee3\u8c22\u578b\u7b5b\u9009\u6807\u8bb0(\u6269\u589e\u578b\u57fa\u56e0\u7b5b\u9009\u6807\u8bb0)<\/p>\n<\/section>\n<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"35.0000%\">\n<section>\n<section>\u4e8c\u6c22\u53f6\u9178\u8fd8\u539f\u9176\uff08DHFR\uff09<\/section>\n<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"34.9900%\">\n<section>\n<section>\u7532\u6c28\u8776\u5464\uff08MTX\uff09<\/section>\n<\/section>\n<\/td>\n<\/tr>\n<tr>\n<td colspan=\"1\" rowspan=\"1\" width=\"35.0000%\">\n<section>\n<section>\u8c37\u6c28\u9170\u80fa\u5408\u6210\u9176\uff08GS\uff09<\/section>\n<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"34.9900%\">\n<section>\n<section>\u6c28\u57fa\u4e9a\u781c\u86cb\u6c28\u9178\uff08MSX\uff09<\/section>\n<\/section>\n<\/td>\n<\/tr>\n<tr>\n<td colspan=\"1\" rowspan=\"5\" width=\"30.0000%\">\n<section>\n<section>\u6297\u751f\u7d20\u7b5b\u9009\u6807\u8bb0(\u975e\u6269\u589e\u578b\u57fa\u56e0\u7b5b\u9009\u6807\u8bb0)<\/p>\n<\/section>\n<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"35.0000%\">\n<section>\n<section>\u560c\u5464\u9709\u7d20\u4e59\u9170\u8f6c\u79fb\u9176\uff08PAC\uff09<\/section>\n<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"34.9900%\">\n<section>\n<section>\u560c\u5464\u9709\u7d20\uff08PD\uff09<\/section>\n<\/section>\n<\/td>\n<\/tr>\n<tr>\n<td colspan=\"1\" rowspan=\"1\" width=\"35.0000%\">\n<section>\n<section>\u6740\u7a3b\u761f\u83cc\u7d20\u8131\u6c28\u9176\uff08BSD\uff09<\/section>\n<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"34.9900%\">\n<section>\n<section>\u6740\u7a3b\u761f\u83cc\u7d20\uff08blasticidin\uff09<\/section>\n<\/section>\n<\/td>\n<\/tr>\n<tr>\n<td colspan=\"1\" rowspan=\"1\" width=\"35.0000%\">\n<section>\n<section>\u6f6e\u9709\u7d20\u78f7\u9178\u8f6c\u79fb\u9176\uff08Hpt\uff09<\/section>\n<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"34.9900%\">\n<section>\n<section>\u6f6e\u9709\u7d20\uff08hygromycin\uff09<\/section>\n<\/section>\n<\/td>\n<\/tr>\n<tr>\n<td colspan=\"1\" rowspan=\"1\" width=\"35.0000%\">\n<section>\u535a\u83b1\u9709\u7d20\u6297\u6027\u57fa\u56e0\uff08G418\uff09<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"34.9900%\">\n<section>\u535a\u83b1\u9709\u7d20\uff08BLM\uff09<\/section>\n<\/td>\n<\/tr>\n<tr>\n<td colspan=\"1\" rowspan=\"1\" width=\"35.0000%\">\n<section>\u6c28\u57fa\u7cd6\u7518\u78f7\u9178\u8f6c\u79fb\u9176\uff08APH\uff09<\/section>\n<\/td>\n<td colspan=\"1\" rowspan=\"1\" width=\"34.9900%\">\n<section>\u65b0\u9709\u7d20\uff08Pnb\uff09<\/section>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<ul>\n<li>\u8f6c\u67d3\u65b9\u6cd5<\/li>\n<\/ul>\n<p>\u8f6c\u67d3\u7c7b\u578b\u4e00\u822c\u6709\u7a33\u5b9a\u8f6c\u67d3\u548c\u77ac\u65f6\u8f6c\u67d3\u4e24\u79cd\u3002\u5728\u5355\u6297\u7ec6\u80de\u682a\u7b5b\u9009\u7a33\u5b9a\u8f6c\u67d3\u524d\u901a\u5e38\u901a\u8fc7\u77ac\u65f6\u8f6c\u67d3\u5f97\u5230\u4e00\u4e9b\u6297\u4f53\uff0c\u8fdb\u884c\u8d28\u91cf\u5206\u6790\uff0c\u4ee5\u521d\u6b65\u5224\u65ad\u6297\u4f53\u662f\u5426\u6ee1\u8db3\u9700\u8981\uff0c\u6216\u8005\u8fdb\u884c\u4e00\u4e9b\u524d\u671f\u7684\u542f\u52a8\u5b50\u3001\u5bc6\u7801\u5b50\u7b49\u7ec4\u4ef6\u7684\u4f18\u5316\u3002\u7a33\u5b9a\u8f6c\u67d3\u4e2d\u5916\u6e90DNA\u65e2\u53ef\u4ee5\u6574\u5408\u5230\u5bbf\u4e3b\u67d3\u8272\u4f53\u4e2d\uff0c\u4e5f\u53ef\u80fd\u4f5c\u4e3a\u4e00\u79cd\u6e38\u79bb\u4f53\u5b58\u5728\u3002\u5728CHO\u7ec6\u80de\u4e2d\u8fdb\u884c\u7a33\u5b9a\u8f6c\u67d3\u65f6\uff0c\u5916\u6e90DNA\u6574\u5408\u5230CHO\u67d3\u8272\u4f53\u4e0a\uff0c\u672a\u6574\u5408\u7684\u6e38\u79bb\u6001\u7684DNA\u4f1a\u968f\u4f20\u4ee3\u800c\u4e22\u5931\u3002<\/p>\n<p>\u5e38\u7528\u4e8e\u54fa\u4e73\u52a8\u7269\u7ec6\u80de\u8f6c\u67d3\u7684\u65b9\u6cd5\u6709\u78f7\u9178\u9499\u6cd5\u3001\u9633\u79bb\u5b50\u8102\u8d28\u4f53\u6cd5\u3001\u9633\u79bb\u5b50\u805a\u5408\u7269\u6cd5\u548c\u7535\u7a7f\u5b54\u6cd5\uff0c\u4f7f\u7528\u6700\u591a\u7684\u662f\u9633\u79bb\u5b50\u8102\u8d28\u4f53\u6cd5\u548c\u7535\u7a7f\u5b54\u6cd5\u3002\u78f7\u9178\u9499\u6cd5\u662f\u501f\u52a9DNA-\u78f7\u9178\u9499\u590d\u5408\u7269\u6c89\u6dc0\u9ecf\u9644\u5728\u7ec6\u80de\u819c\u8868\u9762\uff0c\u901a\u8fc7\u7ec6\u80de\u7684\u5185\u541e\u4f5c\u7528\u800c\u4f7fDNA\u88ab\u7ec6\u80de\u6355\u83b7\u3002\u6c89\u6dc0\u9897\u7c92\u7684\u5927\u5c0f\u548c\u8d28\u91cf\u5bf9\u4e8e\u8f6c\u67d3\u7684\u6210\u529f\u81f3\u5173\u91cd\u8981\uff0c\u4e5f\u53d7pH\u503c\u3001\u9499\u79bb\u5b50\u6d53\u5ea6\u3001DNA\u6d53\u5ea6\u3001\u6c89\u6dc0\u53cd\u5e94\u65f6\u95f4\u3001\u7ec6\u80de\u5b75\u80b2\u65f6\u95f4\u7b49\u56e0\u7d20\u5f71\u54cd\u3002\u78f7\u9178\u9499\u6cd5\u53ef\u9002\u7528\u4e8e\u7a33\u8f6c\u77ac\u8f6c\uff0c\u64cd\u4f5c\u7b80\u4fbf\u82b1\u8d39\u76f8\u5bf9\u8f83\u4f4e\uff0c\u4f46\u8f6c\u67d3\u6548\u7387\u4f4e\uff0c10-20%\uff0c\u5e76\u4e14\u91cd\u590d\u6027\u5f88\u5dee\u3002\u9633\u79bb\u5b50\u8102\u8d28\u4f53\u6cd5\u4f7f\u7528\u5e26\u6b63\u7535\u7684\u8102\u8d28\u4f53\u4e0e\u5e26\u8d1f\u7535\u7684\u6838\u9178\u78f7\u9178\u57fa\u56e2\u5f62\u6210DNA-\u9633\u79bb\u5b50\u8102\u8d28\u4f53\u590d\u5408\u7269\uff0c\u518d\u4e0e\u7ec6\u80de\u819c\u4e0a\u7684\u553e\u6db2\u9178\u6b8b\u57fa\u7684\u8d1f\u7535\u6838\u7ed3\u5408\uff0c\u53ef\u80fd\u7ecf\u8fc7\u5185\u541e\u4f5c\u7528\u88ab\u5bfc\u5165\u7ec6\u80de\u3002\u8be5\u65b9\u6cd5\u4f7f\u7528\u7b80\u5355\uff0c\u53ef\u643a\u5e26\u5927\u7247\u6bb5DNA\uff0c\u901a\u7528\u4e8e\u5404\u79cd\u7c7b\u578b\u7684\u88f8\u9732DNA\u6216RNA\uff0c\u80fd\u8f6c\u67d3\u5404\u79cd\u7c7b\u578b\u7684\u7ec6\u80de\uff0c\u8f6c\u67d3\u6548\u7387\u3001\u8f6c\u67d3\u7684\u7a33\u5b9a\u6027\u548c\u53ef\u91cd\u590d\u6027\u5927\u5927\u63d0\u9ad8\u3002\u4f46\u8f6c\u67d3\u65f6\u9700\u8981\u53bb\u9664\u8840\u6e05\uff0c\u8840\u6e05\u7684\u7f3a\u5931\u4f1a\u5bfc\u81f4\u7ec6\u80de\u6bd2\u6027\u589e\u52a0\uff0c\u8f6c\u67d3\u6548\u679c\u4e5f\u968f\u7ec6\u80de\u7c7b\u578b\u53d8\u5316\u8f83\u5927\u3002\u9633\u79bb\u5b50\u805a\u5408\u7269\u6cd5\u4f7f\u7528\u5e26\u6b63\u7535\u7684\u805a\u5408\u7269\u4e0e\u6838\u9178\u5e26\u8d1f\u7535\u7684\u78f7\u9178\u57fa\u56e2\u5f62\u6210\u5e26\u6b63\u7535\u7684\u590d\u5408\u7269\u540e\u4e0e\u7ec6\u80de\u8868\u9762\u5e26\u8d1f\u7535\u7684\u86cb\u767d\u591a\u7cd6\u76f8\u4e92\u4f5c\u7528\uff0c\u5e76\u901a\u8fc7\u5185\u541e\u4f5c\u7528\u8fdb\u5165\u7ec6\u80de\u3002\u9633\u79bb\u5b50\u805a\u5408\u7269\u6cd5\u5177\u6709\u9633\u79bb\u5b50\u8102\u8d28\u4f53\u7684\u8f6c\u67d3\u6548\u7387\u9ad8\uff0c\u64cd\u4f5c\u7b80\u5355\uff0c\u9002\u7528\u8303\u56f4\u5e7f\uff0c\u91cd\u590d\u6027\u597d\u7b49\u7279\u70b9\u5916\uff0c\u8fd8\u5177\u6709\u5728\u4f53\u5185\u8f6c\u67d3\u6548\u7387\u9ad8\uff0c\u7ec6\u80de\u6bd2\u6027\u4f4e\u7b49\u7279\u70b9\u3002\u7535\u7a7f\u5b54\u6cd5\uff0c\u4e5f\u5c31\u662f\u7535\u8f6c\uff0c\u65e0\u8bba\u662f\u5728\u64cd\u4f5c\u65b9\u6cd5\u3001\u9002\u7528\u8303\u56f4\u4ee5\u53ca\u8f6c\u67d3\u6548\u7387\u7b49\u65b9\u9762\uff0c\u90fd\u6709\u7740\u5f88\u5927\u7684\u4f18\u52bf\u3002\u800c\u73b0\u5b9e\u4ea6\u662f\u5982\u6b64\uff0c\u7531\u4e8e\u7535\u8f6c\u7684\u9ad8\u6548\u3001\u4f4e\u5185\u6bd2\u3001\u64cd\u4f5c\u7b80\u5355\uff0c\u5e76\u4e14\u53ef\u4ee5\u77ac\u65f6\u548c\u7a33\u5b9a\u5730\u8868\u8fbe\u5916\u6e90\u57fa\u56e0\uff0c\u56e0\u6b64\u5e38\u5e38\u7528\u4e8e\u79d1\u7814\u9886\u57df\u7684\u65b0\u836f\u5f00\u53d1\u3001\u764c\u75c7\u7814\u7a76\u3001\u514d\u75ab\u5b66\u7814\u7a76\u7b49\u3002<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-7255 aligncenter\" src=\"https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/0905-1.png\" alt=\"\" width=\"736\" height=\"629\" srcset=\"https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/0905-1.png 372w, https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/0905-1-300x256.png 300w\" sizes=\"auto, (max-width: 736px) 100vw, 736px\" \/><\/p>\n<ul>\n<li>\u9ad8\u8868\u8fbe\u7ec6\u80de\u682a\u7684\u7b5b\u9009<\/li>\n<\/ul>\n<p>\u7b5b\u9009\u9ad8\u8868\u8fbe\u5355\u514b\u9686\u7ec6\u80de\u682a\u662f\u7ec6\u80de\u682a\u6784\u5efa\u7684\u5173\u952e\u6b65\u9aa4\uff0c\u8861\u91cf\u6807\u51c6\u5305\u62ec\u6297\u4f53\u8868\u8fbe\u91cf\u3001\u4ea7\u54c1\u8d28\u91cf\u3001\u4ee3\u8c22\u7a33\u5b9a\u6027\u3001\u7ec6\u80de\u7a33\u5b9a\u6027\u7b49\u3002\u5f53\u76ee\u7684\u57fa\u56e0\u88ab\u8f6c\u67d3\u5230\u5bbf\u4e3b\u7ec6\u80de\u5185\uff0c\u5e76\u6309\u7167\u4e00\u5b9a\u7684\u7ec6\u80de\u5bc6\u5ea6\u5728\u5408\u9002\u7684\u7b5b\u9009\u538b\u529b\u4e0b\u8fdb\u884c\u4f20\u4ee3\u57f9\u517b\u540e\u4fbf\u5f97\u5230\u4e86\u7ec6\u80de\u6c60\u3002\u7ec6\u80de\u6c60\u7684\u8868\u8fbe\u91cf\u53d6\u51b3\u4e8e\u8f6c\u67d3\u7684\u65b9\u5f0f\u3001\u7b5b\u9009\u538b\u529b\u7684\u4f5c\u7528\u4ee5\u53ca\u76ee\u7684\u57fa\u56e0\u6574\u5408\u60c5\u51b5\u3002\u7531\u4e8e\u76ee\u7684\u57fa\u56e0\u6574\u5408\u5177\u6709\u5f88\u5927\u7684\u968f\u673a\u6027\uff0c\u5373\u4f7f\u7528\u76f8\u540c\u7684\u65b9\u6cd5\u8fdb\u884c\u8f6c\u67d3\u548c\u7b5b\u9009\uff0c\u8868\u8fbe\u91cf\u4f9d\u7136\u4f1a\u4f53\u73b0\u51fa\u6781\u5927\u7684\u5dee\u5f02\u6027\uff0c\u63a5\u4e0b\u6765\u5c31\u8981\u4ece\u7ec6\u80de\u6c60\u4e2d\u7b5b\u9009\u51fa\u9ad8\u4ea7\u7a33\u5b9a\u7684\u7ec6\u80de\u682a\u3002<\/p>\n<p>CHO\u7ec6\u80de\u8f6c\u67d3\u4e4b\u540e\uff0c\u901a\u5e38\u4f1a\u505a\u4e00\u4e2a\u514b\u9686\u6c60(pool)\uff0c\u7136\u540e\u8fdb\u884c\u7b5b\u9009\u3002\u7b5b\u9009\u7b56\u7565\u4e3b\u8981\u6d89\u53ca\u6297\u751f\u7d20\u6216\u836f\u7269\u7684\u4ee3\u8c22\u9014\u5f84\uff0c\u5e38\u89c1\u7684\u7b5b\u9009\u8bd5\u5242\u5305\u62ecpuromycin\u3001G418\u3001MTX\u548cMSX\u7b49\u3002Puromycin\u4e3a\u6c28\u57fa\u7cd6\u7519\u7c7b\u6297\u751f\u7d20\uff0c\u901a\u8fc7\u5e72\u6270\u6838\u7cd6\u4f53\u529f\u80fd\u963b\u65ad\u54fa\u4e73\u52a8\u7269\u7ec6\u80de\u7684\u86cb\u767d\u8d28\u5408\u6210\uff0c\u6765\u81ea\u94fe\u9709\u83cc\u7684pac\u57fa\u56e0\u5177\u6709\u89e3\u9664\u560c\u5464\u9709\u7d20(puromycin)\u6bd2\u6027\u7684\u4f5c\u7528\uff0c\u5728\u7b5b\u9009\u7684\u65f6\u5019\uff0cpuromycin\u6d53\u5ea6\u4e00\u822c\u572810\uff5e50ug\/ml\uff1bG418\u4e5f\u662f\u4e00\u79cd\u6c28\u57fa\u7cd6\u82f7\u7c7b\u6297\u751f\u7d20\uff0c\u662f\u7a33\u5b9a\u8f6c\u67d3\u6700\u5e38\u7528\u7684\u6297\u6027\u7b5b\u9009\u8bd5\u5242\u4e4b\u4e00\uff0cG418\u5728\u7b5b\u9009\u7684\u65f6\u5019\u4e00\u822c\u6d53\u5ea6\u8303\u56f4\u4e3a200\uff5e1000ug\/ml\uff1bMTX\u4e3a\u53f6\u9178\u62ee\u6297\u5242\uff0c\u5728\u7ec6\u80de\u5185\u7ecf\u8fc7\u8f6c\u6362\u540e\u53ef\u6291\u5236DHFR\u7684\u6d3b\u6027\uff0c\u6291\u5236\u6838\u9178\u5408\u6210\uff0c\u5f15\u8d77\u7ec6\u80de\u6bd2\u6027\u3002Schimke\u7b49\u7684\u7814\u7a76\u663e\u793a\uff0c\u968f\u7740MTX\u6d53\u5ea6\u7684\u589e\u52a0\uff0c\u7edd\u5927\u591a\u6570\u7ec6\u80de\u6b7b\u4ea1\uff0c\u4f46\u5728\u6781\u5c11\u6570\u5e78\u5b58\u4e0b\u6765\u7684\u6297\u6027\u7ec6\u80de\u4e2d\uff0cDHFR\u57fa\u56e0\u5f97\u4ee5\u6269\u589e\uff0c\u76ee\u7684\u57fa\u56e0\u62f7\u8d1d\u6570\u968f\u4e4b\u589e\u52a0\uff0c\u63d0\u9ad8\u4e86\u8868\u8fbe\u91cf\u3002MTX\u7b5b\u9009\u65f6\uff0c\u4e00\u822c\u6d53\u5ea6\u8303\u56f4\u4e3a25\uff5e1000 nmol\/L\uff1bMSX\u7b5b\u9009\u91c7\u7528\u7684\u662f\u8c37\u6c28\u9170\u80fa\u5408\u6210\u9176\u57fa\u56e0GS\u7cfb\u7edf\u538b\u529b\uff0c\u8c37\u6c28\u9170\u80fa\u5408\u6210\u9176(GS)\u6269\u589e\u7cfb\u7edf\u662f\u65b0\u8fd1\u53d1\u5c55\u7684\u66f4\u6709\u6548\u7684\u7cfb\u7edf\uff0c\u5177\u6709\u66f4\u9ad8\u7684\u6269\u589e\u6548\u5e94\u3002MSX\u7b5b\u9009\u65f6\uff0c\u4e00\u822c\u6d53\u5ea6\u8303\u56f4\u4e3a25\uff5e500 umol\/L\u3002<\/p>\n<p><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-7256 aligncenter\" src=\"https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/0905-2.jpg\" alt=\"\" width=\"716\" height=\"477\" srcset=\"https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/0905-2.jpg 1280w, https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/0905-2-300x200.jpg 300w, https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/0905-2-1030x686.jpg 1030w, https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/0905-2-768x512.jpg 768w, https:\/\/chimebiologics.com\/wp-content\/uploads\/2024\/09\/0905-2-705x470.jpg 705w\" sizes=\"auto, (max-width: 716px) 100vw, 716px\" \/><\/p>\n<p>\u5e38\u7528\u7684\u7b5b\u9009\u514b\u9686\u7684\u65b9\u6cd5\u5305\u62ec\uff1a\u6709\u9650\u7a00\u91ca\u6cd5\uff08limiting dilution cloning\uff0cLDC\uff09\u3001\u6d41\u5f0f\u7ec6\u80de\u4eea\u5206\u9009\u6cd5\uff08fluorescence activated cell sorter\uff0cFACS\uff09\u3001\u5355\u7ec6\u80de\u6253\u5370\u6280\u672f\u3001\u534a\u56fa\u4f53\u57f9\u517b\u57fa\u7b5b\u9009\u6cd5\u7b49\u3002<\/p>\n<p>\u6709\u9650\u7a00\u91ca\u6cd5<\/p>\n<p>\u6709\u9650\u7a00\u91ca\u6cd5\u56e0\u4e3a\u5176\u4f4e\u6210\u672c\u548c\u6613\u4e8e\u64cd\u4f5c\uff0c\u66fe\u5e7f\u6cdb\u8fd0\u7528\u4e8e\u514b\u9686\u7684\u7b5b\u9009\u3002\u8be5\u6cd5\u662f\u5c06\u7ec6\u80de\u7a00\u91ca\u5230\u6781\u4f4e\u7684\u7ec6\u80de\u5bc6\u5ea6\uff0c\u5e76\u5c06\u7a00\u91ca\u597d\u7684\u7ec6\u80de\u7f6e\u4e8e96\u5b54\u677f\u4e2d\u57f9\u517b\uff0c\u4f7f\u5b54\u677f\u4e2d\u6bcf\u4e2a\u5b54\u7684\u7406\u8bba\u7ec6\u80de\u6570\u5c0f\u4e8e1\u4e2a\uff0c\u7a00\u91ca\u540e\u4f7f\u7528\u663e\u5fae\u955c\u5bf996\u5b54\u677f\u6574\u677f\u62cd\u7167\uff0c\u4ee5\u786e\u4fdd\u5355\u514b\u9686\u3002\u7ecf\u8fc7\u4e00\u6bb5\u65f6\u95f4\u7684\u57f9\u517b\u540e\u4f7f\u7528\u9176\u8054\u514d\u75ab\u5438\u9644\u53cd\u5e94\u9009\u53d6\u8868\u8fbe\u91cf\u9ad8\u7684\u7ec6\u80de\u6269\u5927\u57f9\u517b\u3002\u6709\u9650\u7a00\u91ca\u6cd5\u5f80\u5f80\u9700\u8981\u91cd\u590d\u4e24\u6b21\uff0c\u4ee5\u786e\u4fdd\u5355\u514b\u9686\u7ec6\u80de\u682a\u7684\u7eaf\u5ea6\u3002<\/p>\n<p>\u6d41\u5f0f\u7ec6\u80de\u4eea\u5206\u9009\u6cd5<\/p>\n<p>\u5c06\u5e26\u6709\u8367\u5149\u6807\u8bb0\u7684\u4e8c\u6297\u548c\u5206\u6ccc\u6297\u4f53\u7684CHO\u7ec6\u80de\u6df7\u5408\u5b75\u80b2\uff0c\u5206\u6ccc\u5230\u7ec6\u80de\u8868\u9762\u7684\u6297\u4f53\u5c31\u80fd\u591f\u88ab\u6d41\u5f0f\u7ec6\u80de\u4eea\u68c0\u6d4b\u5230\uff0c\u4ece\u800c\u5229\u7528\u5176\u5206\u9009\u529f\u80fd\u7b5b\u9009\u51fa\u5206\u6ccc\u591a\u7684\u7ec6\u80de\u3002\u4e3a\u4e86\u4f7f\u5206\u6ccc\u5230\u80de\u5916\u7684\u6297\u4f53\u80fd\u591f\u7ef4\u6301\u5728\u7ec6\u80de\u819c\u8868\u9762\uff0c\u8fd8\u53ef\u4ee5\u4f7f\u7528\u5fae\u56ca\u5c06\u7ec6\u80de\u4ee5\u53ca\u5206\u6ccc\u7684\u6297\u4f53\u5305\u88f9\u8d77\u6765\u3002\u76ee\u524d\uff0c\u53c8\u6709\u65b0\u7684\u65b9\u6cd5\u8fd0\u7528\u4e8e\u6d41\u5f0f\u7ec6\u80de\u4eea\u5206\u9009\uff0c\u5373\u5c06\u7eff\u8272\u8367\u5149\u86cb\u767d\uff08green fluorescent 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