{"id":24970,"date":"2020-05-26T15:30:35","date_gmt":"2020-05-26T07:30:35","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=24970"},"modified":"2025-03-21T14:24:49","modified_gmt":"2025-03-21T06:24:49","slug":"immortalized-human-epidermal-keratinocytes-sik-%e8%b2%a8%e8%99%9ft0770","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=24970","title":{"rendered":"Immortalized Human Epidermal Keratinocytes (SIK) | \u8ca8\u865fT0770"},"content":{"rendered":"<p><strong>A<\/strong><strong>bmgood<\/strong><strong>\u85e5\u7269\u7be9\u9078\u5e73\u53f0<\/strong><strong> | <\/strong><strong>\u7d30\u80de\u7cfb\u5217\u5546\u54c1<\/strong><strong> (immortalized cell)<\/strong><\/p>\n<p><span style=\"color: #00ccff;\"><a style=\"color: #00ccff;\" href=\"http:\/\/biopioneer.com.tw\/?p=24970\">Immortalized Human Epidermal Keratinocytes (SIK) | \u8ca8\u865fT0770\u00a0<\/a><\/span> \u7d30\u80de\u6578\u91cf1&#215;10<sup>6<\/sup> cells \/ 1.0 ml<\/p>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"https:\/\/www.abmgood.com\/pub\/media\/catalog\/product\/cache\/926507dc7f93631a094422215b778fe0\/t\/0\/t0770-fig2.jpg\" alt=\"\" width=\"466\" height=\"478\" \/><\/p>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"https:\/\/www.abmgood.com\/pub\/media\/catalog\/product\/cache\/926507dc7f93631a094422215b778fe0\/t\/0\/t0770-fig2.jpg\" alt=\"\" width=\"415\" height=\"426\" \/><strong>\u7522\u54c1\u4ecb\u7d39:<\/strong>\u6c38\u751f\u5316\u7684\u4eba\u8868\u76ae\u89d2\u8cea\u5f62\u6210\u7d30\u80de\uff08Human Epidermal Keratinocytes , SIK\uff09\u4f86\u6e90\u65bc\u4eba\u7684foreskin\u6a23\u54c1\uff0c\u9023\u7e8c\u57f9\u990a\u7684\u89d2\u8cea\u5f62\u6210\u7d30\u80de\u5c0e\u81f4\u5176\u81ea\u767c\u6c38\u751f\u5316(spontaneous immortalization)\u3002\u6c38\u751f\u7684\u7d30\u80de\u985e\u4f3c\u65bc\u6b63\u5e38\u7684\u4eba\u8868\u76ae\u83cc\u843d\uff0c\u4f46\u662f\u7d30\u80de\u9031\u671f\u8abf\u7bc0\u86cb\u767d\uff08\u4f8b\u5982\u7d30\u80de\u9031\u671f\u86cb\u767dA\uff09\u7684\u6c34\u5e73\u6025\u5287\u589e\u52a0\u3002\u7528TGF-f\u6216TPA\u8655\u7406\u6c38\u751f\u7684\u4eba\u8868\u76ae\u89d2\u8cea\u5f62\u6210\u7d30\u80de\uff08SIK\uff09\u5728\u9632\u6b62\u4f4e\u9223\u57f9\u990a\u57fa\u4e2d\u751f\u9577\u65b9\u9762\u975e\u5e38\u6709\u6548\u8207\u6b63\u5e38\u7684\u89d2\u8cea\u5f62\u6210\u7d30\u80de\u76f8\u4f3c\u3002\u901a\u904e\u5728\u8868\u76ae\u751f\u9577\u56e0\u5b50\uff08EGF\uff09\u7684\u5b58\u5728\u4e0b\u57f9\u990a\uff0c\u6c38\u751f\u7d30\u80de\u4e2d\u7684\u7aef\u7c92\u9176\u6d3b\u6027\u88ab\u523a\u6fc0\u4e86\u7d045\u500d\u3002\u6c38\u751f\u5316\u7684\u4eba\u985e\u8868\u76ae\u89d2\u8cea\u5f62\u6210\u7d30\u80de\uff08SIK\uff09\u53ef\u7528\u65bc\u5c08\u6ce8\u65bc\u76ae\u819a\u75c5\u5b78\u7684\u7814\u7a76\uff0c\u7279\u5225\u662f\u5728\u76ae\u819a\u81f4\u764c\u4f5c\u7528\u65b9\u9762\uff0c\u56e0\u70ba\u5df2\u767c\u73fe\u4eba\u985e\u76ae\u819a\u4e2d\u7684\u7aef\u7c92\u9176\u6d3b\u5316\u8207\u7d2b\u5916\u7dda\u7167\u5c04\u6709\u95dc\u3002<img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"https:\/\/www.abmgood.com\/pub\/media\/catalog\/product\/cache\/926507dc7f93631a094422215b778fe0\/t\/0\/t0770-fig3.jpg\" alt=\"\" width=\"638\" height=\"489\" \/><\/p>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"https:\/\/www.abmgood.com\/pub\/media\/catalog\/product\/cache\/926507dc7f93631a094422215b778fe0\/t\/0\/t0770-fig3.jpg\" alt=\"\" width=\"712\" height=\"546\" \/><\/p>\n<table width=\"559\">\n<tbody>\n<tr>\n<td width=\"219\"><strong>SKU<\/strong><\/td>\n<td data-th=\"SKU\">T0770<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Species<\/strong><\/td>\n<td data-th=\"Species\">Human<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Tissue\/Organ\/Organ System<\/strong><\/td>\n<td data-th=\"Tissue\/Organ\/Organ System\">Reproductive<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Cell Morphology<\/strong><\/td>\n<td data-th=\"Cell Morphology\">Polygonal<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Applications<\/strong><\/td>\n<td data-th=\"Applications\">For Research Use Only<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Seeding Density<\/strong><\/td>\n<td data-th=\"Seeding Density\">Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Donor Gender<\/strong><\/td>\n<td data-th=\"Donor Gender\">Male<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Donor Gender<\/strong><\/td>\n<td data-th=\"Donor Gender\">Male<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Immortalization Method<\/strong><\/td>\n<td data-th=\"Immortalization Method\">Spontaneous immortalization<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Cell Type<\/strong><\/td>\n<td data-th=\"Cell Type\">Immortalized Cells<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Growth Properties<\/strong><\/td>\n<td data-th=\"Growth Properties\">Adherent<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><\/td>\n<td data-th=\"Propagation Requirements\"><\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>Unit quantity<\/strong><\/td>\n<td data-th=\"Unit quantity\">1&#215;10<sup>6<\/sup>\u00a0cells \/ 1.0 ml<\/td>\n<\/tr>\n<tr>\n<td width=\"219\"><strong>QC<\/strong><\/td>\n<td data-th=\"QC\">1) Chromosomal analysis; 2) Immunoblotting of cell cycle control proteins; 3) Telomerase activity using telomeric repeat amplification protocol<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>Human Epidermal Keratinocytes , SIK\u7d30\u80de\u682a | \u4e0d\u6747\u65b9\u6cd5 : Spontaneous immortalization | \u5668\u5b98\u4f86\u6e90foreskin | \u7d30\u80de\u57f9\u990a\u689d\u4ef6: \u4f7f\u7528PriCoat\u2122T25\u57f9\u990a\u74f6 (<span style=\"color: #008000;\"><a style=\"color: #008000;\" href=\"http:\/\/biopioneer.com.tw\/?p=7046\">\u8ca8\u865fG299<\/a><\/span>)\uff0c\u4ee5\u6700\u4f73\u5316\u521d\u4ee3\u548c\u4e0d\u673d\u7d30\u80de\u7684\u751f\u9577\u3001\u589e\u6b96\u548c\u64f4\u589e\u3002\u6216\u5857\u5c64\u7d30\u80de\u5916\u57fa\u8cea(Applied Cell Extracellular Matrix) \u589e\u5f37\u7d30\u80de\u8cbc\u9644 (<span style=\"color: #008000;\"><a style=\"color: #008000;\" href=\"http:\/\/biopioneer.com.tw\/?p=7031\">\u8ca8\u865fG422<\/a><\/span>)\u3002Grow cells in ECM-coated culture vessels with the following conditions. \u57f9\u990a\u6db2<u>PriGrowVI (<\/u><u>\u8ca8\u865f<\/u><u>\u00a0<\/u><span style=\"color: #008000;\"><u>TM004<\/u><\/span><u>)<\/u>\u3002\u70ba\u4e86\u88fd\u5099\u5b8c\u6574\u7684\u751f\u9577\u57f9\u990a\u57fa\uff0c\u5c07\u4e0b\u5217\u7d44\u5206\u6dfb\u52a0\u5230\u57fa\u790e\u57f9\u990a\u57fa\u4e2d\uff1a\u80ce\u725b\u8840\u6e05\uff08TM999\uff09\u81f3\u6700\u7d42\u6fc3\u5ea65%\u548cPenicillin\/Streptomycin\u9752\u9ef4\u7d20\/\u93c8\u9ef4\u7d20\u6eb6\u6db2\uff08G255\uff09\u81f3\u6700\u7d42\u6fc3\u5ea6\u70ba1\uff05\u3002\u4e8c\u6c27\u5316\u78b3\uff08CO2\uff09\uff1a5\uff05\uff0c\u6eab\u5ea6\uff1a37.0\u2103\u3002<\/p>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"https:\/\/www.abmgood.com\/pub\/media\/catalog\/product\/cache\/926507dc7f93631a094422215b778fe0\/t\/0\/t0770-fig6.jpg\" alt=\"\" width=\"386\" height=\"416\" \/><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"https:\/\/www.abmgood.com\/pub\/media\/catalog\/product\/cache\/926507dc7f93631a094422215b778fe0\/t\/0\/t0770-fig6.jpg\" alt=\"\" width=\"376\" height=\"405\" \/><\/p>\n<p>\u5c07kinase domain\u6fc0\u9176\u7d50\u69cb\u57df\u878d\u5408\u81f3homodimerization domain\u540c\u6e90\u4e8c\u805a\u5316\u7d50\u69cb\u57df\uff08ETV6\u7684PNT domain\u7d50\u69cb\u57df\uff09\u53ef\u6fc0\u6d3b\u6fc0\u9176kinase\u3002 SYK Stable Ba \/ F3\u7d30\u80de\u682a\u53ef\u8868\u73fe\u6d3b\u5316\u7684SYK\u916a\u6c28\u9178\u6fc0\u9176\u4e26\u5177GFP\u5831\u5c0e\u86cb\u767d\u8868\u73fe\uff08GFP-ETV6-SYK\u67b6\u69cb\u65bcSED2\u8f09\u9ad4\uff09\u3002 \u8868\u9054\u6d3b\u5316\u7684\u916a\u6c28\u9178\u6fc0\u9176\u7684Ba \/ F3\u7d30\u80de\u5b8c\u5168\u4f9d\u8cf4\u65bc\u6fc0\u9176\u6d3b\u6027\u7684\u5b58\u6d3b\uff0c\u56e0\u6b64\u5373\u4f7f\u5728\u4e0d\u5b58\u5728IL-3\u7684\u60c5\u6cc1\u4e0b\u4e5f\u53ef\u4ee5\u7e41\u6b96\u7d30\u80de\u3002This cell line carries the SED2 vector which does not have any selection markers<\/p>\n<p><span style=\"color: #3366ff;\"><a style=\"color: #3366ff;\" href=\"http:\/\/biopioneer.com.tw\/?p=5545\">\u9ef4\u6f3f\u83ccPCR\u5075\u6e2c<\/a>\u00a0<strong>|<\/strong>\u00a0<a style=\"color: #3366ff;\" href=\"http:\/\/biopioneer.com.tw\/?p=5582\">DAPI\u67d3\u6838<\/a>\u00a0<strong>|<\/strong>\u00a0<a style=\"color: #3366ff;\" href=\"http:\/\/biopioneer.com.tw\/?p=5564\">\u7d30\u80de\u5167\u9ef4\u6f3f\u83cc\u53bb\u9664\u5291<\/a>\u00a0<strong>|<\/strong>\u00a0<a style=\"color: #3366ff;\" href=\"http:\/\/biopioneer.com.tw\/?p=5567\">\u6c34\u6d74\u69fd\u6291\u83cc\u5291<\/a>\u00a0<strong>|<\/strong>\u00a0<a style=\"color: #3366ff;\" href=\"http:\/\/biopioneer.com.tw\/?p=5575\">\u5674\u9727\u5f0f\u53bb\u9ef4\u5291 \u7b49\u7cfb\u5217\u5546\u54c1<\/a><\/span><\/p>\n<p><strong>Abmgood<\/strong><strong>\u4e0d\u673d\u5316\u7cfb\u5217\u5546\u54c1<\/strong>\u00a0<strong>(immortalized reagenet)<\/strong>\u00a0Ready-to-use<\/p>\n<p>Abmgood\u516c\u53f8\u65bc\u4eba\u985e\u8207\u8001\u9f20\u4e0d\u673d\u4e4b\u7d30\u80de\u682a\u4e0a\uff0c\u5177\u6709\u5e7e\u5341\u5e74\u7684\u5be6\u9a57\u7d93\u9a57\uff1b\u63d0\u4f9b\u4e00\u7cfb\u5217\u4e0d\u673d (immortalized)\u5316\u4e4b\u7d30\u80de\u682a\u3001\u53ca\u4e0d\u673d\u5316\u4e4b\u8a66\u5291\u5546\u54c1\u3002ready-to-use\u5b9a\u91cf\u597d\u4e4bretrovirus\u3001lentiviru\u3001 adenovirus\uff0c\u53ef\u76f4\u63a5\u52a0\u65bc\u7d30\u80de\u4e2d\u9032\u884c\u611f\u67d3\u8868\u73fe\uff1b\u75c5\u6bd2\u6db2\u5305\u62ec\u6709SV40\u91cd\u7d44\u75c5\u6bd2\u3001EBV\u91cd\u7d44\u75c5\u6bd2\u3001hTERT\u75c5\u6bd2\u6db2\u3001Myc\u57fa\u56e0\u75c5\u6bd2\u6db2\u3001p53\u57fa\u56e0\u75c5\u6bd2\u6db2\u3001Rb\u57fa\u56e0\u75c5\u6bd2\u6db2\u3001Ras\u57fa\u56e0\u75c5\u6bd2\u6db2\u3002\u63d0\u4f9b106\u53ca109\u5169\u7a2e\u5305\u88dd\u529b\u50f9\u4e4b\u75c5\u6bd2\u6db2\uff1b\u4ea6\u63d0\u4f9b\u4e00\u7cfb\u5217\u75c5\u6bd2\u529b\u50f9\u6fc3\u7e2e\u8207\u5b9a\u91cf\u8a66\u5291\u5546\u54c1\u3002<\/p>\n<p>\u7d30\u80de\u4e0d\u673d\u5316\u7684\u5de5\u5177\u4e2d\uff0cSV40 T antigen\u662f\u6700\u7c21\u55ae\u4e14\u6700\u5e38\u7528\u4f86\u9020\u6210\u4e0d\u673d\u5316\u7d30\u80de\u751f\u6210\u7684\u65b9\u6cd5\u3002\u591a\u6578\u7684\u75c5\u6bd2\u57fa\u56e0\u548c\u6291\u764c\u57fa\u56e0\uff08p53\u3001Rb\u7b49\uff09\u7684\u4e0d\u6d3b\u5316\u76f8\u95dc\uff0c\u6703\u8a98\u5c0e\u7d30\u80de\u8655\u65bcreplicative senescence\u7684\u968e\u6bb5\u3002\u76ee\u524d\u4e5f\u6709\u7814\u7a76\u986f\u793a\u88ab\u611f\u67d3\u7684\u7d30\u80de\uff0cSV40 T anbtigen\u6703\u8a98\u5c0e\u7aef\u7c92\u9176\uff08Telomerase\uff09\u7684\u6d3b\u6027\u3002\u8207\u7aef\u7c92\uff08telomere\uff09\u9577\u5ea6\u76f8\u95dc\u7684Telomerase Reverse Transcriptase protein (TERT)\u662f\u4e00\u7a2e\u4e0d\u673d\u7d30\u80de\u7684\u7814\u7a76\u61c9\u7528\u3002\u5e73\u5e38\u7d30\u80de\u4e2d\uff0chTERT\u662f\u8655\u65bc\u4e0d\u6d3b\u5316\u614b\uff0c\u5fc5\u8981\u6642\u6703\u9032\u884c\u6d3b\u5316\u4fdd\u6301telomere\u7684\u9577\u5ea6\uff0c\u907f\u514d\u7d30\u80de\u767c\u751freplicative senescence\u7684\u73fe\u8c61\u3002\u67d0\u4e9b\u7d30\u80dehTERT \u904e\u5ea6\u8868\u73fe\uff08over-expression\uff09\u5c0d\u65bc\u8a98\u767c\u7d30\u80de\u4e0d\u673d\u5316\u662f\u7121\u6548\u7684\uff08\u5982\u521d\u4ee3\u4e0a\u76ae\u7d30\u80de\uff09\uff0c\u53cd\u800c\u6703\u9020\u6210\u7d30\u80de\u6b7b\u4ea1\uff1b\u540c\u6642\u8868\u73fehTERT catalytic subunit\u4ee5\u53cap53\u6216RB siRNA\u5728\u4eba\u985e\u521d\u4ee3\u5375\u5de2\u4e0a\u76ae\u7d30\u80de\u6703\u9020\u6210\u7d30\u80de\u4e0d\u673d\u5316\u3002\u904e\u5ea6\u8868\u73feRas\u6216Myc T58A mutants\u4e5f\u6709\u521d\u4ee3\u7d30\u80de\u4e0d\u673d\u5316\u7684\u73fe\u8c61\u7522\u751f\u3002<\/p>\n<p><strong>Decomplement<\/strong><strong>\u52a0\u71b1\u975e\u6d3b\u5316<\/strong><strong>FBS<\/strong><strong>\uff1a<\/strong><\/p>\n<p>1.\u8b93FBS\u74f6\u57284\u00b0C\u4e0b\u5b8c\u5168\u89e3\u51cd\u904e\u591c\u3002<\/p>\n<p>2.\u5c07FBS\u74f6\u7f6e\u65bc56\u00b0C\u7684\u6c34\u6d74\u4e2d30\u5206\u9418\u3002<\/p>\n<p>Decomplemented FBS\u53ef\u4ee5\u5728-20\u00b0C\u4e0b\u9577\u671f\u4fdd\u5b58\u3002\u907f\u514d\u983b\u7e41\u7684\u51cd\u878d\u5faa\u74b0\u3002\u6bcf2-3\u5929\u66f4\u63db\u4e00\u6b21\u5b8c\u6574\u7684\u5a92\u9ad4\u3002\u4e0d\u8981\u8b93\u4ecb\u8cea\u984f\u8272\u8b8a\u70ba\u9ec3\u8272\u3002<\/p>\n<p><strong>Thawing<\/strong><strong>\u7d30\u80de\u89e3\u51cd\u65b9\u6cd5\uff1a<\/strong><strong>(<\/strong><strong>\u6bcf\u7a2e\u7d30\u80de\u5efa\u8b70\u689d\u4ef6\u53ef\u80fd\u6709\u8a31\u4e0d\u540c\uff0c\u4f9d\u7167\u539f\u5ee0\u5efa\u8b70\u64cd\u4f5c\uff0c\u4ee5\u4e0b\u70ba\u901a\u7528\u65b9\u5f0f<\/strong><strong>)<\/strong><\/p>\n<p>1.\u572837\u2103\u6c34\u6d74\u4e2d\u89e3\u51cd\u5c0f\u74f6\uff0c\u76f4\u5230\u53ea\u6709\u4e00\u5c0f\u584a\u51b0\u584a\u3002<\/p>\n<p>2.\u5c07\u7d30\u80de\u8f49\u79fb\u5230\u542b\u67095ml pre-warmed complete media \u4e4b15ml\u7ba1\u4e2d\u3002<\/p>\n<p>3.\u5c07\u7d30\u80de\u4ee5290xg\u96e2\u5fc35\u5206\u9418\u3002<\/p>\n<p>4.\u5c0f\u5fc3\u4e1f\u68c4\u4e0a\u6e05\u6db2\uff0c\u4e0d\u8981\u6253\u64fe\u7d30\u80de\u6c89\u6fb1\uff0c\u8f15\u8f15\u5730\u4e0a\u4e0b\u79fb\u6db2\uff0c\u5c07\u7d30\u80de\u91cd\u65b0\u61f8\u6d6e\u57281ml\u5b8c\u5168\u57f9\u990a\u57fa\u4e2d\u3002<\/p>\n<p>5.\u5c07\u7d30\u80de\u57f9\u990a\u5728150,000-200,000\u500b\u7d30\u80de\/ cm2\u3002<\/p>\n<p><strong>Subculture<\/strong><strong>\u7d30\u80de\u7e7c\u4ee3\u65b9\u6cd5<\/strong><strong>:<\/strong>\u00a0<strong>(<\/strong><strong>\u6bcf\u7a2e\u7d30\u80de\u5efa\u8b70\u689d\u4ef6\u53ef\u80fd\u6709\u8a31\u4e0d\u540c\uff0c\u4f9d\u7167\u539f\u5ee0\u5efa\u8b70\u64cd\u4f5c\uff0c\u4ee5\u4e0b\u70ba\u901a\u7528\u65b9\u5f0f<\/strong><strong>)<\/strong><\/p>\n<p>1.\u7576\u7d30\u80de70-90\uff05 confluency\u6642\uff0c\u5efa\u8b70\u7e7c\u4ee3\u57f9\u990a\u3002<\/p>\n<ol start=\"2\">\n<li>Aspirate old media\u3002\u61f8\u6d6e\u6db2\u4e2d\u7684\u4e00\u4e9b\u7d30\u80de\u4ecd\u7136\u662f\u6d3b\u7d30\u80de\u3002\u6216\u8005\uff0c\u4e5f\u53ef\u4ee5\u6536\u96c6\u542b\u6709\u61f8\u6d6e\u7d30\u80de\u7684\u4e0a\u6e05\u6db2\u4e26\u96e2\u5fc3\uff08Skip to Step 6 in protocol\uff09\u3002<\/li>\n<\/ol>\n<p>3.\u52a0\u51651\uff1a1\u6bd4\u4f8b\u76841X\u7121\u83ccPBS\u548c0.25\uff05\u80f0\u86cb\u767d\u9176-EDTA\uff08TM051\uff09\u3002<\/p>\n<p>4.\u5728\u5ba4\u6eab\u4e0b\u5b75\u80b2\u7d30\u80de3-5\u5206\u9418\u4e26\u652a\u62cc\u57f9\u990a\u5bb9\u5668\u76f4\u81f390\uff05\u7684\u7d30\u80de\u812b\u843d\u3002<\/p>\n<ol start=\"5\">\n<li>\u4e2d\u548c\u80f0\u86cb\u767d\u9176:\u6dfb\u52a0\u7b49\u91cf\u4e4btrypsin + PBS\u3002<\/li>\n<\/ol>\n<p>6.\u6536\u96c6\u7d30\u80de\u4e26\u4ee5290xg\u96e2\u5fc35\u5206\u9418\u3002<\/p>\n<p>7.\u4e1f\u68c4\u4e0a\u6e05\u6db2\uff0c\u8f15\u8f15\u5730\u4e0a\u4e0b\u79fb\u6db2\uff0c\u5c07\u7d30\u80de\u91cd\u65b0\u61f8\u6d6e\u5728\u5b8c\u5168\u57f9\u990a\u57fa\u4e2d\u3002<\/p>\n<p>8.\u5efa\u8b70split ratio\u6bd4\u4e0d\u8d85\u904e1\uff1a3\u3002<\/p>\n<p><strong>Freezing<\/strong><strong>\u7d30\u80de\u51cd\u7d50\uff1a<\/strong><strong>(<\/strong><strong>\u6bcf\u7a2e\u7d30\u80de\u5efa\u8b70\u689d\u4ef6\u53ef\u80fd\u6709\u8a31\u4e0d\u540c\uff0c\u4f9d\u7167\u539f\u5ee0\u5efa\u8b70\u64cd\u4f5c\uff0c\u4ee5\u4e0b\u70ba\u901a\u7528\u65b9\u5f0f<\/strong><strong>)<\/strong><\/p>\n<p>Complete growth medium with decomplemented FBS to a final concentration of 50% from PAN Biotech (P40-37500) or from Nucleus Biologics (FBS1824-001) and 5% DMSO.<\/p>\n<p>\u5132\u5b58\u6eab\u5ea6\uff1a\u6db2\u6c2e\u6c23\u76f8\u3002\u91cd\u8981\u63d0\u793a\uff1a\u6b64\u7d30\u80de\u4fc2\u5c0dFBS\u6279\u6b21\u975e\u5e38\u654f\u611f\u3002\u6211\u5011\u5f37\u70c8\u5efa\u8b70\u6700\u7d42\u7528\u6236\u4f7f\u7528\u5f9ePAN biotech\u6216Nucleus Biologics \u4e4bFBS\u3002<\/p>\n<p>\u6162\u75c5\u6bd2\u5305\u88dd\u7cfb\u5217\u7522\u54c1\u00a0<span style=\"color: #008080;\"><a style=\"color: #008080;\" href=\"http:\/\/biopioneer.com.tw\/?p=6146\"><u>Lentivirus<\/u><u>\u6162\u75c5\u6bd2\u4f4e\u901f\u7d14\u5316<\/u>(\u7d14\u5316)\u00a0<\/a>| <a style=\"color: #008080;\" href=\"http:\/\/biopioneer.com.tw\/?p=6152\"><u>Lentivirus <\/u><u>\u6162\u75c5\u6bd2\u9ad8\u6fc3\u5ea6\u7d14\u5316<\/u>(\u7d14\u5316)\u00a0<\/a>|\u00a0<a style=\"color: #008080;\" href=\"http:\/\/biopioneer.com.tw\/?p=6156\"><u>Lentivirus<\/u><u>\u75c5\u6bd2<\/u><u>RT-QPCR<\/u><u>\u5b9a\u91cf<\/u>(\u5b9a\u91cf)<\/a>\u00a0<\/span><\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Abmgood\u85e5\u7269\u7be9\u9078\u5e73\u53f0 | \u7d30\u80de\u7cfb\u5217\u5546\u54c1 (immortalized cell) Immortalized [&hellip;]<\/p>\n","protected":false},"author":3,"featured_media":11653,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[1],"tags":[1524],"_links":{"self":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/24970"}],"collection":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=24970"}],"version-history":[{"count":4,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/24970\/revisions"}],"predecessor-version":[{"id":50791,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/24970\/revisions\/50791"}],"wp:featuredmedia":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/media\/11653"}],"wp:attachment":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=24970"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=24970"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=24970"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}