{"id":22478,"date":"2019-11-08T17:18:43","date_gmt":"2019-11-08T09:18:43","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=22478"},"modified":"2025-08-14T11:41:24","modified_gmt":"2025-08-14T03:41:24","slug":"%e6%b0%b8%e7%94%9f%e5%8c%96%e7%9a%84%e4%ba%ba%e6%a4%8e%e9%96%93%e7%9b%a4%e8%bb%9f%e9%aa%a8%e7%b5%82%e6%9d%bf%e4%be%86%e6%ba%90%e7%9a%84%e5%b9%b9%e7%b4%b0%e8%83%9e-sv40-immortalized-human-interverteb","status":"publish","type":"post","link":"https:\/\/biopioneer.com.tw\/?p=22478","title":{"rendered":"\u6c38\u751f\u5316\u7684\u4eba\u690e\u9593\u76e4\u8edf\u9aa8\u7d42\u677f\u4f86\u6e90\u7684\u5e79\u7d30\u80de-SV40 | Immortalized Human Intervertebral Disc Cartilage Endplate-derived Stem Cells -SV40 \u8ca8\u865fT0009"},"content":{"rendered":"

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

Immortalized Human Intervertebral Disc Cartilage Endplate-derived Stem Cells -SV40 | \u8ca8\u865fT0009 <\/a><\/span>\u7d30\u80de\u6578\u91cf1×106<\/sup> cells \/ 1.0 ml<\/p>\n

\u7522\u54c1\u4ecb\u7d39: \u6c38\u751f\u5316\u7684\u4eba\u690e\u9593\u76e4\u8edf\u9aa8\u7d42\u677f\u4f86\u6e90\u7684\u5e79\u7d30\u80de-SV40(Immortalized Human Intervertebral Disc Cartilage Endplate-derived Stem Cells -SV40) \u662f\u6709\u95dc\u8edf\u9aa8\u7d42\u677f\u548c\u9ad4\u5167\u71df\u990a\u904b\u8f38\u7684\u6709\u50f9\u503c\u4e14\u53ef\u9760\u7684\u4f86\u6e90\u3002\u8edf\u9aa8\u4f4d\u65bc\u810a\u67f1\u7684\u690e\u9aa8\u7d42\u677f\u548c\u9ad3\u6838\u4e4b\u9593\uff0c\u5728\u5176\u4e2d\u5b83\u8d77\u8457\u5c4f\u969c\u7684\u4f5c\u7528\u3002\u5b83\u5728\u5149\u76e4\u7684\u5065\u5eb7\u65b9\u9762\uff08\u4f8b\u5982growth\u751f\u9577\u548cdegeneration\u9000\u5316\uff09\u8d77\u4f5c\u7528\u3002\u6709\u4e86\u6c38\u751f\u5316\u7684\u7d30\u80de\u7cfb\uff0c\u7814\u7a76\u4eba\u54e1\u7121\u9700\u4e0d\u65b7\u5c0b\u627e\u4f9b\u9ad4\u4f86\u5206\u96e2\u7d30\u80de\u3002\u9019\u4e9b\u7d30\u80de\u53ef\u4ee5\u5728\u9ad4\u5916\u589e\u6b96\u4ee5\u7528\u65bc\u4e0b\u6e38\u5be6\u9a57\u3002<\/p>\n

\u7d30\u80de\u7279\u6027 <\/span><\/strong><\/span>\u6c38\u751f\u5316\u7684\u4eba\u690e\u9593\u76e4\u8edf\u9aa8\u7d42\u677f\u4f86\u6e90\u7684\u5e79\u7d30\u80de-SV40 (Immortalized Human Intervertebral Disc Cartilage Endplate-derived Stem Cells -SV40)\u7d30\u80de\u682a | \u4e0d\u6747\u65b9\u6cd5 : Infected with a non-replicating SV40 lentivirus | \u5668\u5b98\u4f86\u6e90Cartilaginous endplate | \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\u6db2PriGrowIV (<\/u>\u8ca8\u865f<\/u>\u00a0<\/u>TM004<\/u><\/span>)<\/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\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<\/p>\n

<\/p>\n

\"\u7db2\u7ad9\u898f\u683c\u5c0f\u5716_\u5de5\u4f5c\u5340\u57df<\/a><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Cat. No.<\/td>\nT0009<\/span><\/td>\n<\/tr>\n
Name<\/td>\nImmortalized Human Intervertebral Disc Cartilage Endplate-Derived Stem Cells – SV40<\/td>\n<\/tr>\n
Description<\/td>\nImmortalized Human Intervertebral Disc Cartilage Endplate-derived Stem Cells -SV40 is a valuable and reliable source for studies relating to the cartilaginous endplate and nutrient transportation in the body. The cartilage is positioned between the vertebral endplate in the spine and the nucleus pulposus in which it functions as a barrier. It has a role in the health aspects of the disc such as growth and degeneration. With an immortalized cell line, researchers do not need to constantly look for donors to isolate the cells. These cells can be proliferated\u00a0in vitro<\/em>\u00a0for downstream experiments.<\/td>\n<\/tr>\n
Organism<\/td>\nHuman (H. sapiens)<\/td>\n<\/tr>\n
Tissue<\/td>\nSkeletal Muscle<\/td>\n<\/tr>\n
Donor History<\/td>\nNot disclosed<\/td>\n<\/tr>\n
Growth Properties<\/td>\nAdherent, polygonal shaped<\/td>\n<\/tr>\n
Cell Type<\/td>\nImmortalized Cells<\/td>\n<\/tr>\n
Storage Condition<\/td>\nVapor phase of liquid nitrogen, or below -130\u00b0C.<\/td>\n<\/tr>\n
Shipping Conditions<\/td>\nShip with dry ice.<\/td>\n<\/tr>\n
Product Format<\/td>\nFrozen<\/td>\n<\/tr>\n
Intended Use<\/td>\nThis product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.<\/td>\n<\/tr>\n
BioSafety<\/td>\nII<\/td>\n<\/tr>\n
Certificate of Analysis<\/td>\nFor batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.<\/td>\n<\/tr>\n
Growth Conditions<\/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. PriGrow IV (TM004<\/a>) + 10% FBS + 1% Penicillin\/Streptomycin Solution (G255<\/a>), 37.0\u00b0C, 5% CO\u2082<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

HOT!!<\/strong><\/span>\u00a0<\/u><\/a><\/span>\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><\/span><\/p>\n

\"abm\u00e5\u0095\u0086\u00e5\u0093\u0081\u00e6\u008f\u0092\u00e5\u009c\u0096\"<\/p>\n

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

\"cell-biology-tab-final<\/p>\n

\u54fa\u4e73\u52d5\u7269\u7d30\u80de\u7684<\/strong>\u4e0d<\/strong>\u673d<\/strong>\u5316Immortalized\u7b56\u7565<\/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><\/td>\nViruses Offered<\/strong><\/td>\nApplication<\/strong><\/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>\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)\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(DMSO\u5c0d\u7d30\u80de\u6709\u6bd2\u6027\uff0cover-thaw\u904e\u5ea6\u89e3\u51cd\u4e0b\u53ef\u80fd\u9020Stress)<\/span><\/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\u572850,000 – 100,000 cells\/cm2<\/sup>\u3002\u00a0cells will have viability of around 40-50%. Post thawing, it will need 4-5 days for cells to recover in culture; Recommended split ratio is 1:4 to 1:5<\/p>\n

\"\"<\/p>\n

DMSO\u5728\u7d30\u80de\u51cd\u5b58\u4e2d\u7684\u61c9\u7528<\/span><\/strong><\/p>\n

\u4e8c\u7532\u57fa\u4e9e\u78b8\u662f\u4e00\u7a2e\u91cd\u8981\u7684\u6ef2\u900f\u578b\u7d30\u80de\u4fdd\u8b77\u5291\u3002\u5728\u6df1\u4f4e\u6eab\uff08\u96f6\u4e0b200\u5ea6\uff09\u4fdd\u5b58\u7d30\u80de\u6642\u51cd\u5b58\u904e\u7a0b\u4e2d\uff0c\u70ba\u9632\u6b62\u7d30\u80de\u5167\u6db2\u51b0\u6676\u5f62\u6210\u3001\u6ef2\u900f\u58d3\u6539\u8b8a\u3001\u7d30\u80de\u7d50\u69cb\u7d0a\u4e82\u7b49\u5c0e\u81f4\u7684\u640d\u50b7\uff0c\u6709\u5fc5\u8981\u4f7f\u7528\u542b\u6709DMSO\u51b7\u51cd\u4fdd\u8b77\u5291\u3002DMSO\u80fd\u5920\u5feb\u901f\u7a7f\u900f\u7d30\u80de\u819c\u9032\u5165\u7d30\u80de\u4e2d\uff0c\u964d\u4f4e\u51b0\u9ede\u3001\u5ef6\u7de9\u51cd\u5b58\u904e\u7a0b\uff0c\u540c\u6642\u63d0\u9ad8\u7d30\u80de\u5167\u96e2\u5b50\u6fc3\u5ea6\uff0c\u6e1b\u5c11\u7d30\u80de\u5167\u51b0\u6676\u7684\u5f62\u6210\uff0c\u5f9e\u800c\u6e1b\u5c11\u7d30\u80de\u640d\u50b7\u3002\u6df1\u4f4e\u6eab\u6642\u4e8c\u7532\u57fa\u4e9e\u78b8\u7684\u7d30\u80de\u6bd2\u6027\u53d7\u5230\u6291\u5236\u3002\u5fa9\u8607\u6642\u52d5\u4f5c\u8981\u5feb\uff0c\u5118\u5feb\u6d17\u6389\u4e8c\u7532\u57fa\u4e9e\u78b8\uff0c\u5426\u5247\u6703\u9020\u6210\u5c0d\u7d30\u80de\u56b4\u91cd\u7684\u6bd2\u6027\u3002\u4e8c\u7532\u57fa\u4e9e\u78b8\uff08DMSO\uff09\u662f\u76ee\u524d\u6700\u597d\u7684\u7d30\u80de\u51cd\u5b58\u4fdd\u8b77\u5291\uff0c\u4f46\u4e5f\u662f\u4e00\u7a2e\u4ee5\u7d30\u80de\u6bd2\u6027\u5f88\u5927\u7684\u5316\u5b78\u8a66\u9a57\u5291\u3002\u7814\u7a76\u7d50\u679c\u8868\u660e\uff0c\u57f9\u990a\u6db2\u4e2dDMSO\u6fc3\u5ea6\u70ba10%\u6642\uff0c\u7d30\u80de\u751f\u9577\u6291\u5236\u7387\u8fd1100%\uff1b1\u2030\u6fc3\u5ea6\u6642\u6291\u5236\u7387\u70ba35%\uff0c\u5373\u4f7f\u662f0.04\u2030\u7684\u6fc3\u5ea6\uff0cDMSO\u5c0d\u7d30\u80de\u7684\u751f\u9577\u4e5f\u6709\u4e0d\u5229\u7684\u5f71\u97ff\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

2. 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<\/p>\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

5. \u4e2d\u548c\u80f0\u86cb\u767d\u9176:\u6dfb\u52a0\u7b49\u91cf\u4e4btrypsin + PBS\u3002<\/p>\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(\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)\u00a0<\/strong>\u64cd\u4f5c\u65b9\u5f0f\u4e0b\u8f09<\/a><\/strong><\/span><\/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

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

Abmgood\u4e0d\u673d\u7cfb\u5217\u5546\u54c1 (immortalized cell) Immortalized Human In […]<\/p>\n","protected":false},"author":3,"featured_media":11653,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[453],"tags":[1833],"_links":{"self":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/22478"}],"collection":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=22478"}],"version-history":[{"count":5,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/22478\/revisions"}],"predecessor-version":[{"id":52352,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/22478\/revisions\/52352"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/media\/11653"}],"wp:attachment":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=22478"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=22478"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=22478"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}