{"id":22736,"date":"2019-12-31T15:20:14","date_gmt":"2019-12-31T07:20:14","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=22736"},"modified":"2025-03-21T14:26:44","modified_gmt":"2025-03-21T06:26:44","slug":"%e4%ba%ba%e5%ad%90%e5%ae%ae%e5%85%a7%e8%86%9c%e7%95%b0%e4%bd%8d%e7%b4%b0%e8%83%9e-immortalized-human-endometriotic-cell-line-12z-%e8%b2%a8%e8%99%9ft0764","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=22736","title":{"rendered":"\u4eba\u5b50\u5bae\u5167\u819c\u7570\u4f4d\u7d30\u80de | Immortalized Human Endometriotic Cell Line (12Z) \u8ca8\u865fT0764"},"content":{"rendered":"

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

Immortalized Human Endometriotic Cell Line (12Z) \u6c38\u751f\u5316\u4eba\u5b50\u5bae\u5167\u819c\u7570\u4f4d\u7d30\u80de\u682a\u00a0 | \u8ca8\u865fT0764 \u7d30\u80de\u6578\u91cf1×106<\/sup> cells \/ 1.0 ml<\/a><\/span><\/strong><\/span><\/p>\n

\"abm\u5546\u54c1\u63d2\u5716\"<\/p>\n

\u7522\u54c1\u4ecb\u7d39:<\/span><\/strong><\/span> \u6c38\u751f\u5316\u7684\u4eba\u985e\u5b50\u5bae\u5167\u819c\u7570\u4f4d\u7d30\u80de\u682a\uff08Immortalized Human Endometriotic Cell Line , 12Z\uff09\u662f\u901a\u904e\u611f\u67d3SV40\u75c5\u6bd2\u611f\u67d3\u7684\uff0c\u9019\u4e9b\u7d30\u80de\u662f\u5f9e\u63a5\u53d7\u8179\u8154\u93e1\u6aa2\u67e5\u768437\u6b72\u5973\u6027\u4e2d\u7372\u5f97\u7684\u3002\u8a72\u7d30\u80de\u682a\u8868\u73fe\u51fa\u9ad4\u5167\u53ef\u898b\u7684\u5b50\u5bae\u5167\u819c\u7570\u4f4d\u75c5\u8b8a\u7684\u6a19\u8a8c\u7269\u7684\u8868\u9054\u3002\u6c38\u751f\u5316\u4eba\u985e\u5b50\u5bae\u5167\u819c\u7570\u4f4d\u7d30\u80de\u682a\uff0812Z\uff09\u53ef\u7528\u65bc\u8868\u5fb5\u548c\u9451\u5b9a\u8207\u5b50\u5bae\u5167\u819c\u7570\u4f4d\u76f8\u95dc\u7684\u57fa\u56e0\u548c\/\u6216\u6a19\u8a18\u3002Immortalized Human Endometriotic Cell Line (12Z) was generated by infecting cells, obtained from a 37 year-old female undergoing laparoscopy, with SV40 virus. This cell line exhibits expression of markers of endometriotic lesions that are seen in vivo. Immortalized Human Endometriotic Cell Line (12Z) may be used to characterize and identify genes and\/or markers related to endometriosis.<\/p>\n

\u5fc5\u9700\u8cfc\u8cb7PriCoat\u2122T25\u57f9\u990a\u74f6 (\u8ca8\u865fG299<\/a><\/span>)\uff0c\u57f9\u990a\u6db2PriGrowIII (<\/u>\u8ca8\u865f<\/u>\u00a0<\/u>TM003<\/u>)<\/u><\/span>\u3002<\/p>\n

\"\"<\/p>\n

Immortalized Human Endometriotic Cell Line\u00a0<\/strong>\uff08SV40 virus\uff09<\/p>\n\n\n\n\n\n\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>\nUterus<\/td>\n<\/tr>\n
Donor Age<\/strong><\/td>\n37 years old<\/td>\n<\/tr>\n
Donor Gender<\/strong><\/td>\nFemale<\/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>\nFibroblast-like<\/td>\n<\/tr>\n
Recommended Seeding Density<\/strong><\/td>\n15,000 – 20,000 cells\/cm2<\/sup><\/td>\n<\/tr>\n
Population Doubling<\/strong><\/td>\n55 – 65 hours<\/td>\n<\/tr>\n
Immortalization Method<\/strong><\/td>\nImmortalized with SV40 virus<\/td>\n<\/tr>\n
Description<\/strong><\/td>\nImmortalized Human Endometriotic Cell Line (12Z) was generated by infecting cells, obtained from a 37 year-old female undergoing laparoscopy, with SV40 virus. This cell line exhibits expression of markers of endometriotic lesions that are seen\u00a0in vivo<\/em>. Immortalized Human Endometriotic Cell Line (12Z) may be used to characterize and identify genes and\/or markers related to endometriosis.<\/td>\n<\/tr>\n
Depositor Information<\/strong><\/td>\nGoethe University<\/td>\n<\/tr>\n
Applications<\/strong><\/td>\nFor Research Use Only<\/td>\n<\/tr>\n
Markers<\/strong><\/td>\nSV40T, cytokeratin, vimentin, N-cadherin, estrogen receptor \u03b1 and \u03b2, progesterone receptor, and aromatase P450<\/td>\n<\/tr>\n
Propagation Requirements<\/strong><\/td>\nUse of PriCoat\u2122 T25 Flasks (G299<\/a>) or Applied Cell Extracellular Matrix (G422<\/a>) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions.\u00a0The base medium for this cell line is Prigrow III medium available at\u00a0abm, Cat. No.\u00a0TM003<\/a>. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999<\/a>)* to a final concentration of 10%, and Penicillin\/Streptomycin Solution (G255<\/a>) to a final concentration of 1%.
\nCarbon dioxide (CO2<\/sub>): 5%, Temperature: 37.0\u00b0C.
\n* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Immortalized Human Endometriotic Cell Line , 12Z\u7d30\u80de\u682a | \u4e0d\u6747\u65b9\u6cd5 : Immortalized with SV40 virus | \u5668\u5b98\u4f86\u6e90Uterus | \u7d30\u80de\u57f9\u990a\u689d\u4ef6: \u4f7f\u7528PriCoat\u2122T25\u57f9\u990a\u74f6 (\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 (\u8ca8\u865fG422<\/a><\/span>)\u3002Grow cells in ECM-coated culture vessels with the following conditions. \u57f9\u990a\u6db2PriGrowIII (<\/u>\u8ca8\u865f<\/u>\u00a0<\/u>TM003<\/u>)<\/u><\/span>\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\u5ea610\uff05\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>\n

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

\"ABM\u7d30\u80de\u57f9\u990a_\u5de5\u4f5c\u5340\u57df<\/p>\n

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

A<\/strong>bmgood<\/strong>\u4e0d<\/strong>\u673d<\/strong>\u5316\u7cfb\u5217\u5546\u54c1<\/strong>\u00a0(immortalized cell)<\/strong><\/span><\/p>\n

\u6839\u64da\u7d30\u80de\u985e\u578b\uff0cPrimary cells\u521d\u4ee3\u7d30\u80de\u5728\u9ad4\u5916\u5177\u6709\u5c0f\u65bc10-50\u6b21\u50b3\u4ee3\u7684\u6709\u9650\u58fd\u547d\u3002 \u4e0d\u673d\u5316\u7d30\u80de\u682a\u80fd\u5920\u5ef6\u9577\u589e\u6b96\uff0c\u4e26\u4e14\u5177\u6709\u8207\u89aa\u4ee3\u7d44\u7e54\u76f8\u4f3c\u6216\u76f8\u540c\u7684\u57fa\u56e0\u578b\u548c\u8868\u73fe\u578b\u3002<\/p>\n

\u54fa\u4e73\u52d5\u7269\u7d30\u80de\u7684\u4e0d\u673d\u5316Immortalized\u7b56\u7565<\/span><\/strong><\/span><\/p>\n

\u5b58\u5728\u7528\u65bc\u5728\u57f9\u990a\u7269\u4e2d\u4e0d<\/strong>\u673d<\/strong>\u5316<\/strong>\u54fa\u4e73\u52d5\u7269\u7d30\u80de\u7684\u5e7e\u7a2e\u65b9\u6cd5\u3002 Abmgood\u591a\u5e74\u7814\u767c\u7d93\u9a57\uff0c\u958b\u767c\u51fa\u4e86\u4e00\u7cfb\u5217Immortalized\u5546\u54c1\uff1b\u5305\u62ec\u5373\u7528\u578b\u9006\u8f49\u9304\u75c5\u6bd2\uff0c\u6162\u75c5\u6bd2\u548c\u817a\u75c5\u6bd2\u8f09\u9ad4\u3002<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Gene Overexpressed<\/strong><\/span><\/td>\nViruses Offered<\/strong><\/span><\/td>\nApplication<\/strong><\/span><\/td>\n<\/tr>\n
hTERT<\/a><\/td>\nAdenovirus,
\nLentivirus,
\nRetrovirus<\/td>\n		\u53ef\u9760\u548c\u5e38\u7528\u3002 \u53ef\u80fd\u5728\u4e00\u4e9b\u7d30\u80de\u985e\u578b\uff08\u4f8b\u5982\u4e0a\u76ae\u7d30\u80de\uff09\u4e2d\u5f15\u8d77\u7d30\u80de\u6b7b\u4ea1<\/td>\n<\/tr>\n
SV40 T Antigen<\/a><\/td>\nAdenovirus,
\nLentivirus,
\nRetrovirus<\/td>\n		\u53ef\u9760\u548c\u5e38\u7528\u3002 \u901a\u5e38\u7528\u65bc\u4e0d\u673d\u5316\u4e0a\u76ae\u7d30\u80de\u3002<\/td>\n<\/tr>\n
EBV genes<\/a><\/td>\nEpstein-Barr Virus<\/td>\n\u4e0d\u673d\u5316B\u6dcb\u5df4\u7d30\u80de\uff0c\u6709\u6642\u662fT\u6dcb\u5df4\u7d30\u80de\u3002<\/td>\n<\/tr>\n
HPV E6-E7<\/a><\/td>\nLentivirus<\/td>\n\u4e0d\u673d\u5316\u89d2\u5316\u7d30\u80de\u3002<\/td>\n<\/tr>\n
HOXB8, HOXA9, HOXA10<\/a><\/td>\nLentivirus<\/td>\n\u4e0d\u673d\u5316\u9020\u8840\u7d30\u80de\uff0c\u5305\u62ec\u5de8\u566c\u7d30\u80de\uff0c\u9020\u8840\u7956\u7d30\u80de\u548c\u9aa8\u9ad3\u7956\u7d30\u80de\u3002<\/td>\n<\/tr>\n
Adenoviral E1A-E1B<\/a><\/td>\nLentivirus<\/td>\n\u4e0d\u673d\u5316\u5927\u7bc4\u570d\u7684\u521d\u7d1a\u56d3\u9f52\u52d5\u7269\u7d30\u80de\uff0c\u5305\u62ec\u7f8a\u6c34\u7d30\u80de\uff0c\u539f\u767c\u6027\u4e0a\u76ae\u7d30\u80de\uff0c\u6210\u7e96\u7dad\u7d30\u80de\u548c\u539f\u4ee3\u814e\u7d30\u80de\u3002<\/td>\n<\/tr>\n
p53, Rb siRNA<\/a><\/td>\nLentivirus<\/td>\n\u8207hTERT\u5171\u8868\u9054\u4ee5\u66f4\u597d\u5730\u4f7f\u4e0a\u76ae\u7d30\u80de\u4e0d\u673d\u5316\u3002<\/td>\n<\/tr>\n
Bmi1<\/a><\/td>\nLentivirus<\/td>\n\u4e0d\u673d\u5316\u4e0a\u76ae\u7d30\u80de\u3002<\/td>\n<\/tr>\n
CDK4<\/a><\/td>\nLentivirus<\/td>\n\u8207hTERT\u5171\u540c\u8868\u9054\u66f4\u771f\u5be6\u7684\u7d30\u80de\u6a21\u578b\u3002<\/td>\n<\/tr>\n
Myc T58A (c-Myc), Ras V12 (H-Ras)<\/a><\/td>\nLentivirus<\/td>\n\u4e0d\u673d\u5316\u5404\u7a2e\u7d30\u80de\uff0c\u5305\u62ec\u795e\u7d93\u5e79\u7d30\u80de\uff0c\u521d\u4ee3\u814e\u7d30\u80de\uff0c\u6210\u7d05\u7d30\u80de\u548c\u5404\u7a2e\u4e0a\u76ae\u7d30\u80de\uff08\u539f\u767c\u6027\u524d\u5217\u817a\uff0c\u4e73\u817a\uff0c\u6c23\u9053\u7b49\uff09\u3002<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Decomplement<\/strong>\u52a0\u71b1\u975e\u6d3b\u5316FBS\uff1a<\/strong><\/span><\/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><\/span>\u7d30\u80de\u89e3\u51cd\u65b9\u6cd5\uff1a(\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)<\/span>\u00a0<\/strong>\u64cd\u4f5c\u65b9\u5f0f\u4e0b\u8f09<\/strong><\/a><\/span><\/span><\/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>\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>\u00a0\u64cd\u4f5c\u65b9\u5f0f\u4e0b\u8f09<\/strong><\/a><\/span><\/span><\/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><\/span>\u7d30\u80de\u51cd\u7d50\uff1a(\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)<\/span>\u00a0<\/strong><\/span>\u64cd\u4f5c\u65b9\u5f0f\u4e0b\u8f09<\/a><\/strong><\/span><\/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

      \u6162\u75c5\u6bd2\u5305\u88dd\u7cfb\u5217\u7522\u54c1Lentivirus<\/u>\u6162\u75c5\u6bd2\u4f4e\u901f\u7d14\u5316<\/u>(\u7d14\u5316)\u00a0<\/a>|\u00a0Lentivirus\u00a0<\/u>\u6162\u75c5\u6bd2\u9ad8\u6fc3\u5ea6\u7d14\u5316<\/u>(\u7d14\u5316)\u00a0<\/a>|\u00a0Lentivirus<\/u>\u75c5\u6bd2RT-QPCR<\/u>\u5b9a\u91cf<\/u>(\u5b9a\u91cf)<\/a>\u00a0<\/span><\/p>\n","protected":false},"excerpt":{"rendered":"

      Abmgood\u85e5\u7269\u7be9\u9078\u5e73\u53f0 | \u7d30\u80de\u7cfb\u5217\u5546\u54c1 (immortalized cell) Immortalized […]<\/p>\n","protected":false},"author":3,"featured_media":11653,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[453],"tags":[1767],"_links":{"self":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/22736"}],"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=22736"}],"version-history":[{"count":5,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/22736\/revisions"}],"predecessor-version":[{"id":50796,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/22736\/revisions\/50796"}],"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=22736"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=22736"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=22736"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}