{"id":21418,"date":"2019-07-23T11:07:46","date_gmt":"2019-07-23T03:07:46","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=21418"},"modified":"2022-07-20T11:06:35","modified_gmt":"2022-07-20T03:06:35","slug":"immortalized-human-aortic-endothelial-cells-sv40-abmgood%e8%b2%a8%e8%99%9ft0447","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=21418","title":{"rendered":"\u4e3b\u52d5\u8108\u5167\u76ae\u7d30\u80de-SV40 |Immortalized Human Aortic Endothelial Cells- SV40 | ABMGOOD\u8ca8\u865fT0447"},"content":{"rendered":"

A<\/strong>bmgood<\/strong>\u85e5\u7269\u7be9\u9078\u5e73\u53f0<\/strong> | <\/strong>\u7d30\u80de\u7cfb\u5217\u5546\u54c1<\/strong> (immortalized cell)<\/strong><\/p>\n

Immortalized Human Aortic Endothelial Cells- SV40<\/strong>\u00a0 | \u8ca8\u865fT0447 <\/a><\/span>\u7d30\u80de\u6578\u91cf1×106<\/sup> cells \/ 1.0 ml<\/p>\n

\u7522\u54c1\u4ecb\u7d39: \u6c38\u751f\u5316\u4eba\u4e3b\u52d5\u8108\u5167\u76ae\u7d30\u80de-SV40 (Immortalized Human Aortic Endothelial Cells- SV40) \u3002\u4e0d\u6747\u65b9\u6cd5 : Serial passaging and transduction with recombinant lentiviruses carrying SV40 Large T antigen | \u5668\u5b98\u4f86\u6e90Aorta | \u7d30\u80de\u57f9\u990a\u689d\u4ef6: \u57fa\u790e\u57f9\u990a\u6db2\u00a0PriGrowIII (<\/u>\u8ca8\u865f<\/u>\u00a0<\/u>TM003<\/u>)<\/u><\/a><\/span>\u3002\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\u5ea610\uff05\uff0c\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\u9664\u975e\u53e6\u6709\u8aaa\u660e\uff0c\u4e0d\u5efa\u8b70\u4f7f\u7528\u71b1\u6ec5\u6d3b\u7684FBS( \u00a0heat-inactivated FBS) \u9032\u884c\u7d30\u80de\u57f9\u990a\u3002*\u8acb\u6ce8\u610f\uff0c\u7d30\u80de\u751f\u9577\u7de9\u6162\uff0c\u53ef\u80fd\u9700\u8981\u9577\u90541\u9031\u624d\u80fd\u5f9e\u89e3\u51cd\u4e2d\u6062\u5fa9\u3002<\/p>\n

\"\"<\/p>\n

Immortalized Human Aortic Endothelial Cells- SV40<\/strong>\uff08GFP-ETV6-SYK\/SED2 Vector\uff09<\/p>\n\n\n\n\n\n\n\n\n\n\n\n
Organism<\/strong><\/td>\nHomo sapiens<\/td>\n<\/tr>\n
Source Organ<\/strong><\/td>\nAorta<\/td>\n<\/tr>\n
BioSafety Level<\/strong><\/td>\nII<\/td>\n<\/tr>\n
Growth Properties<\/strong><\/td>\nAdherent<\/td>\n<\/tr>\n
Morphology<\/strong><\/td>\nEpithelial (<10% fibroblasts)<\/td>\n<\/tr>\n
Recommended Seeding Density<\/strong><\/td>\nThaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
\n* Note that cells are slow growing and may take up to 1 week to recover from thaw.<\/strong><\/td>\n<\/tr>\n
Population Doubling<\/strong><\/td>\n40-50 hours<\/td>\n<\/tr>\n
Immortalization Method<\/strong><\/td>\nSerial passaging and transduction with recombinant lentiviruses carrying SV40 Large T antigen<\/td>\n<\/tr>\n
Applications<\/strong><\/td>\nFor Research Use Only<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u9ef4\u6f3f\u83ccPCR\u5075\u6e2c<\/a>\u00a0|<\/strong>\u00a0DAPI\u67d3\u6838<\/a>\u00a0|<\/strong>\u00a0\u7d30\u80de\u5167\u9ef4\u6f3f\u83cc\u53bb\u9664\u5291<\/a>\u00a0|<\/strong>\u00a0\u6c34\u6d74\u69fd\u6291\u83cc\u5291<\/a>\u00a0|<\/strong>\u00a0\u5674\u9727\u5f0f\u53bb\u9ef4\u5291 \u7b49\u7cfb\u5217\u5546\u54c1<\/a><\/span><\/p>\n

\"\"<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Product<\/strong><\/td>\nSize<\/strong><\/td>\nCat. No.<\/strong><\/td>\n<\/tr>\n
Immortalized Human Carotid Artery Smooth Muscle Cells – hTERT<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0511<\/td>\n<\/tr>\n
Immortalized Human Carotid Artery Smooth Muscle Cells – SV40<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0510<\/td>\n<\/tr>\n
Immortalized Human Brachiocephalic Artery Endothelial Cells – SV40<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0513<\/td>\n<\/tr>\n
Immortalized Human Carotid Artery Endothelial Cells – SV40<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0512<\/td>\n<\/tr>\n
Immortalized Human Cardiac Microvascular Endothelial Cells – SV40<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0514<\/td>\n<\/tr>\n
Immortalized Human Aortic Endothelial Cells- SV40<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0447<\/td>\n<\/tr>\n
Immortalized Human Pulmonary Artery Endothelial Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0550<\/td>\n<\/tr>\n
Immortalized Human Coronary Artery Endothelial Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0551<\/td>\n<\/tr>\n
Immortalized Human Internal Thoracic Artery Endothelial Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0552<\/td>\n<\/tr>\n
Immortalized Human Subclavian Artery Endothelial Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0553<\/td>\n<\/tr>\n
Immortalized Human Iliac Artery Endothelial Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0555<\/td>\n<\/tr>\n
Immortalized Human Subclavian Artery Smooth Muscle Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0556<\/td>\n<\/tr>\n
Immortalized Human Coronary Artery Smooth Muscle Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0557<\/td>\n<\/tr>\n
Immortalized Human Pulmonary Artery Smooth Muscle Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0558<\/td>\n<\/tr>\n
Immortalized Human Internal Thoracic Artery Smooth Muscle Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0560<\/td>\n<\/tr>\n
Immortalized Human Brachiocephalic Artery Smooth Muscle Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0561<\/td>\n<\/tr>\n
Immortalized Human Aortic Smooth Muscle Cells – SV40<\/a><\/td>\n1×106<\/sup>\u00a0cells \/ 1.0 ml<\/td>\nT0515<\/td>\n<\/tr>\n
Immortalized Human Ascending Aorta Vascular Smooth Muscle Cells<\/a><\/td>\n1×106<\/sup>\u00a0cells\/ml<\/td>\nT0549<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Abmgood<\/strong>\u4e0d\u673d\u5316\u7cfb\u5217\u5546\u54c1<\/strong>\u00a0(immortalized reagenet)<\/strong>\u00a0Ready-to-use<\/p>\n

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

\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

Decomplement<\/strong>\u52a0\u71b1\u975e\u6d3b\u5316<\/strong>FBS<\/strong>\uff1a<\/strong><\/p>\n

1.\u8b93FBS\u74f6\u57284\u00b0C\u4e0b\u5b8c\u5168\u89e3\u51cd\u904e\u591c\u3002<\/p>\n

2.\u5c07FBS\u74f6\u7f6e\u65bc56\u00b0C\u7684\u6c34\u6d74\u4e2d30\u5206\u9418\u3002<\/p>\n

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

Thawing<\/strong>\u7d30\u80de\u89e3\u51cd\u65b9\u6cd5\uff1a<\/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><\/p>\n

1.\u572837\u2103\u6c34\u6d74\u4e2d\u89e3\u51cd\u5c0f\u74f6\uff0c\u76f4\u5230\u53ea\u6709\u4e00\u5c0f\u584a\u51b0\u584a\u3002<\/p>\n

2.\u5c07\u7d30\u80de\u8f49\u79fb\u5230\u542b\u67095ml pre-warmed complete media \u4e4b15ml\u7ba1\u4e2d\u3002<\/p>\n

3.\u5c07\u7d30\u80de\u4ee5290xg\u96e2\u5fc35\u5206\u9418\u3002<\/p>\n

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

5.\u5c07\u7d30\u80de\u57f9\u990a\u5728150,000-200,000\u500b\u7d30\u80de\/ cm2\u3002<\/p>\n

Subculture<\/strong>\u7d30\u80de\u7e7c\u4ee3\u65b9\u6cd5<\/strong>:<\/strong>\u00a0(<\/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><\/p>\n

1.\u7576\u7d30\u80de70-90\uff05 confluency\u6642\uff0c\u5efa\u8b70\u7e7c\u4ee3\u57f9\u990a\u3002<\/p>\n

    \n
  1. 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

    3.\u52a0\u51651\uff1a1\u6bd4\u4f8b\u76841X\u7121\u83ccPBS\u548c0.25\uff05\u80f0\u86cb\u767d\u9176-EDTA\uff08TM051\uff09\u3002<\/p>\n

    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

      \n
    1. \u4e2d\u548c\u80f0\u86cb\u767d\u9176:\u6dfb\u52a0\u7b49\u91cf\u4e4btrypsin + PBS\u3002<\/li>\n<\/ol>\n

      6.\u6536\u96c6\u7d30\u80de\u4e26\u4ee5290xg\u96e2\u5fc35\u5206\u9418\u3002<\/p>\n

      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

      8.\u5efa\u8b70split ratio\u6bd4\u4e0d\u8d85\u904e1\uff1a3\u3002<\/p>\n

      Freezing<\/strong>\u7d30\u80de\u51cd\u7d50\uff1a<\/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><\/p>\n

      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

      \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>\n","protected":false},"excerpt":{"rendered":"

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