{"id":12937,"date":"2017-08-20T13:01:18","date_gmt":"2017-08-20T05:01:18","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=12937"},"modified":"2022-07-21T11:22:30","modified_gmt":"2022-07-21T03:22:30","slug":"briter-%e7%b4%b0%e8%83%9e%e6%a0%aa%ef%bd%9c-bmp-responsive-immortalized-reporter-briter-cell-line%e8%b2%a8%e8%99%9ft3013","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=12937","title":{"rendered":"BRITER \u7a69\u5b9a\u7d30\u80de\u682a\uff5c BMP Responsive Immortalized Reporter (BRITER) Cell Line\u8ca8\u865fT3015"},"content":{"rendered":"

\u5831\u5c0e\u57fa\u56e0\u7a69\u5b9a\u7d30\u80de\u682a\u5546\u54c1Reporter cell lines<\/span><\/p>\n

Reporter gene\u5831\u5c0e\u57fa\u56e0\u7814\u7a76\uff0c\u5c07\u7d30\u83cc\uff0c\u7d30\u80de\u57f9\u990a\u7269\uff0c\u52d5\u7269\u6216\u690d\u7269\u4e2d\u53e6\u4e00\u7a2e\u611f\u8208\u8da3\u7684\u57fa\u56e0\u8abf\u7bc0\u5e8f\u5217\u67b6\u69cb\u65bc\u5728\u8868\u73fe\u8f09\u9ad4\u4e0a\u3002Abmgood\u63d0\u4f9b\u7cfb\u5217\u5831\u5c0e\u57fa\u56e0\u87a2\u5149\u7d20\u9176\uff0cGFP\uff0cRFP\uff0c\u96d9\u91cd\u5831\u544a\u57fa\u56e0\u548cUTR\uff0c\u7a69\u5b9a\u8868\u73fe\u7b49\u7d30\u80de\u682a\u3002\u87a2\u5149\u7d20\u9176\u901a\u5e38\u7528\u4f5c\u5831\u544a\u57fa\u56e0\uff0c\u4ee5\u8a55\u4f30\u611f\u8208\u8da3\u7684\u555f\u52d5\u5b50\u4e0b\u87a2\u5149\u7d20\u9176\u57fa\u56e0\u7684\u907a\u50b3\u69cb\u5efa\u9ad4\u4e4b\u8f49\u9304\u6d3b\u6027\u3002\u87a2\u5149\u7d20\u9176\u7a69\u5b9a\u7684\u7d30\u80de\u53ef\u7528\u65bc\u52d5\u7269\u6a21\u578b\u4e2d\u7684\u975e\u4fb5\u5165\u6027\u53ef\u8996\u5316\uff0c\u61c9\u7528\u65bc\u751f\u7269\u9ad4\u7684\u7279\u5fb5\u9451\u5b9a\u548c\u6e2c\u91cf\u3002<\/p>\n

\u87a2\u5149\u7d20\u9176\u7d30\u80de<\/span>\u00a0\u00a0<\/span>tCD-Luc2\u7d30\u80de<\/a>\u00a0 |\u00a0PC3-Luc2\u7d30\u80de<\/a>\u00a0|\u00a0\u00a0WM-266-4 -luc2\u7d30\u80de<\/a>\u00a0|\u00a0\u00a0AZ-AHR HepG2\u7d30\u80de<\/a>\u00a0 |\u00a0\u00a0AIZ-AR\u7a69\u5b9a22Rv1<\/a>-Luc2\u7d30\u80de |\u00a0AZ-GR\u7a69\u5b9aHela-luc2<\/a>\u00a0|\u00a0BRITER-Luc2<\/a>\u00a0<\/span><\/p>\n

BMP Responsive Immortalized Reporter (BRITER) Cell Line\uff5c \u8ca8\u865f<\/strong>T3015<\/strong><\/strong><\/a><\/span><\/p>\n

\u9aa8\u5f62\u614b\u767c\u751f\u86cb\u767d\uff08Bone morphogenetic proteins ,BMP\uff09\u53ef\u7528\u4f5c\u6cbb\u7642\u5291\uff0c\u7528\u65bc\u5728bone fracture\u9aa8\u6298\uff0cspinal injury\u810a\u9ad3\u640d\u50b7\u6216surgery\u624b\u8853\u5f8c\u7684\u5275\u50b7\u90e8\u4f4d\u4fc3\u9032\u9aa8\u9abc\u7684\u518d\u751f\uff0c\u4f46\u5176\u6548\u80fd\u5c40\u9650\u65bcsystemic-wide side effects\u5168\u8eab\u6027\u7684\u526f\u4f5c\u7528\uff0c\u5c08\u9580\u7684\u8f38\u9001\u7cfb\u7d71\u7684\u5fc5\u8981\u6027\u4ee5\u53ca\u5316\u5408\u7269\u9ad4\u5167\u534a\u8870\u671f\u77ed\u800c\u6709\u6240\u4e0d\u540c\u3002<\/p>\n

\"\"<\/p>\n

\u751f\u6210BMP\u53cd\u61c9\u6027\u6c38\u751f\u5316\u8a18\u9304\uff08BMP Responsive Immortalized Reporter , BRITER\uff09\u7d30\u80de\uff0c\u9032\u884c\u9ad8\u901a\u91cf\u5316\u5b78\u548c\u5206\u5b50\u907a\u50b3\u4e4b\u7be9\u9078\uff0c\u4ee5\u9451\u5b9aBMP\u4fe1\u865f\u7684\u65b0\u578b\u5c0f\u5206\u5b50\u66ff\u4ee3\u4e4b\u6fc0\u6d3b\u7269\u3002\u7d30\u80de\u662f\u6c38\u751f\u5316\u7684tamoxifen\u4ed6\u83ab\u6614\u82ac,\u53ef\u8a98\u5c0e\u7684Bmp2, Bmp4\u96d9\u91cd\u689d\u4ef6\u6572\u9664\u9f20\u4e4b\u9871\u9aa8\u6210\u9aa8\u7d30\u80de;\u8868\u9054BRE\/\u5177\u87a2\u706b\u87f2\u87a2\u5149\u7d20\u9176\uff08FFLuc\uff09\u57fa\u56e0\u548cSV40\u555f\u52d5\u5b50\/\u589e\u5f37\u5b50\u4f9d\u8cf4\u6027\u6d77\u814e\u87a2\u5149\u7d20\u9176\uff08RRLuc\uff09\u57fa\u56e0\u3002FFLuc\u6d3b\u6027\u65bcBMP\u4fe1\u865f\u50b3\u5c0e\u6d3b\u6027\u800c\u589e\u52a0\uff0c\u800cRRLuc\u57fa\u56e0\u4f5c\u70ba\u5167\u90e8\u5c0d\u7167\uff0c\u4ee5\u9650\u5236\u7531\u975e\u7279\u7570\u6027\u6fc0\u6d3b\u7d30\u80de\u904e\u7a0b\uff08\u4f8b\u5982\u7d30\u80de\u589e\u6b96\uff09\u5f15\u8d77\u7684\u5047\u967d\u6027\u6578\u3002\u6b64\u5916\uff0c\u901a\u904e\u52a0\u51654-hydroxytamoxifen 4-\u7fa5\u57fa\u4ed6\u83ab\u6614\u82ac\u53ef\u4ee5\u589e\u5f37\u5c0d\u5916\u6e90\u6027BMP\u4fe1\u865f\u50b3\u5c0e\u7684\u654f\u611f\u6027\uff0c\u901a\u904e\u52a0\u5165Bog2\u548cBmp4\u57fa\u56e0\u7684Bmp2\u548cBmp4\u57fa\u56e0\u7684\u6572\u9664\u53ef\u964d\u4f4e\u5167\u6e90\u6027BMP\u4fe1\u865f\u7684\u6c34\u5e73\uff0c\u5f9e\u800c\u5c0e\u81f4BRE\u6d3b\u5316\u7684\u7279\u7570\u6027\u548c\u9ad8\u6548\u5831\u544a\u7269<\/p>\n

\"\"<\/p>\n\n\n\n\n\n\n\n\n\n\n
BioSafety Level<\/strong><\/td>\nII<\/td>\n<\/tr>\n
Organism<\/strong><\/td>\nBmp2c<\/sup>\/c<\/sup>;Bmp4c<\/sup>\/c<\/sup>; R26CreER Neonatal Mouse<\/td>\n<\/tr>\n
SourceOrgan<\/strong><\/td>\nCalvariae<\/td>\n<\/tr>\n
Growth Properties<\/strong><\/td>\nAdherent<\/td>\n<\/tr>\n
Morphology<\/strong><\/td>\nMultipolar<\/td>\n<\/tr>\n
Recommended Seeding Density<\/strong><\/td>\nThaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.<\/td>\n<\/tr>\n
Markers<\/strong><\/td>\nOsterix, CollA1, BSP, Runx, Osteocalcin, Alkaline phosphatase<\/td>\n<\/tr>\n
Applications<\/strong><\/td>\nFor Research Use Only<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u00a0<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
Reporter cell lines<\/strong>\u7522\u54c1\u7cfb\u5217<\/strong><\/td>\n\u7d30\u80de\u682a\u8ca8\u865f<\/strong><\/td>\n\u7d30\u80de\u682a\u7269\u7a2e<\/strong><\/td>\n\u5305\u88dd<\/strong><\/td>\n<\/tr>\n
Luciferase (Luc2) Stable TRAMP Cell Line<\/a><\/td>\nT3008<\/td>\nMus musculus<\/td>\n1×106<\/sup> cells\/1.0 ml<\/td>\n<\/tr>\n
Luciferase Stable U87MG\/\u0394EGFR Cell Line<\/a><\/td>\nT3009<\/td>\nHomo sapiens<\/td>\n1×106<\/sup> cells\/1.0 ml<\/td>\n<\/tr>\n
Luciferase Stable PC3 Cell Line<\/a><\/td>\nT3006<\/td>\nHomo sapiens<\/td>\n1×106<\/sup> cells\/1.0 ml<\/td>\n<\/tr>\n
Luciferase Stable WM-266-4 Cell Line<\/a><\/td>\nT3013<\/td>\nHomo sapiens<\/td>\n1×106<\/sup> cells\/1.0 ml<\/td>\n<\/tr>\n
BMP Responsive Immortalized Reporter (BRITER) Cell Line<\/a><\/td>\nT3105<\/td>\nBmp2c<\/sup>\/c<\/sup>;Bmp4c<\/sup>\/c<\/sup>; R26CreER Neonatal Mouse<\/td>\n1×106<\/sup> cells\/1.0ml<\/td>\n<\/tr>\n
AZ-AHR Stable HepG2 Luciferase Reporter Cell Line<\/a><\/td>\nT3102<\/td>\nHomo sapiens<\/td>\n1×106<\/sup> cells\/1.0ml<\/td>\n<\/tr>\n
AZ-GR Stable HeLa Luciferase Reporter Cell Line<\/a><\/td>\nT3103<\/td>\nHomo sapiens<\/td>\n1×106<\/sup> cells\/1.0ml<\/td>\n<\/tr>\n
AIZ-AR Stable 22Rv1 Luciferase Reporter Cell Line<\/a><\/td>\nT3104<\/td>\nHomo sapiens<\/td>\n1×106<\/sup> cells\/1.0ml<\/td>\n<\/tr>\n
HeLa Overexpressing Stable tCD-Luc2 Cell Line<\/a><\/td>\nT6023<\/td>\nHomo sapiens<\/td>\n1×106<\/sup> cells\/1.0ml<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u4e0d\u6764\u5316\u7cfb\u5217\u5546\u54c1 | SV40T Lentivirus\u6162\u75c5\u6bd2\u6db2 <\/a>| hTERT-RFP Lentirus\u75c5\u6bd2\u6db2<\/a> | EBV lentivurus \u6162\u75c5\u6bd2\u6db2<\/a> | Ras V12 lentivirus \u6162\u75c5\u6bd2\u6db2<\/a> | Rb siRNA lentivirus\u6162\u75c5\u6bd2\u6db2<\/a> | p53 siRNA lentivirus\u6162\u75c5\u6bd2\u6db2<\/a> | lentivirus\u75c5\u6bd2\u529b\u50f9\u6fc3\u7e2e\u8207\u5b9a\u91cf\u5546\u54c1<\/a> |<\/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

\"\"<\/p>\n

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

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

\u53ef\u4ee5\u4f7f\u7528PolyIC\uff085\u03bcg\/ ml\uff09\uff0cCpG\uff082mM\uff09\u6216LPS\uff085\u03bcg\/ ml\uff09\u9032\u884c\u523a\u6fc0\u3002\u9019\u4e9b\u7d30\u80de\u5c0dFBS\u7684\u8981\u6c42\u7279\u5225\u654f\u611f\uff0c\u56e0\u6b64\uff0c\u5efa\u8b70\u4f7f\u7528\u76f8\u540c\u7684\u6279\u6b21\u57f9\u990a\u80fd\u5920\u652f\u6301\u5176\u57f9\u990a\u7684\u6700\u4f73\u7d50\u679c\u7684\u7d30\u80de\u3002\u6211\u5011\u9084\u5efa\u8b70\u5728\u5b8c\u6574\u57f9\u990a\u57fa\u4e2d\u6dfb\u52a0\u984d\u5916\u76841\uff05Hepes\u4ee5\u4fc3\u9032\u7d30\u80de\u7e41\u6b96\u3002<\/p>\n

Freezing\u7d30\u80de<\/strong>\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","protected":false},"excerpt":{"rendered":"

\u5831\u5c0e\u57fa\u56e0\u7a69\u5b9a\u7d30\u80de\u682a\u5546\u54c1Reporter cell lines Reporter gene\u5831\u5c0e\u57fa\u56e0\u7814\u7a76\uff0c\u5c07\u7d30\u83cc\uff0c […]<\/p>\n","protected":false},"author":3,"featured_media":11653,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[453],"tags":[3336],"_links":{"self":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/12937"}],"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=12937"}],"version-history":[{"count":4,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/12937\/revisions"}],"predecessor-version":[{"id":18531,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/12937\/revisions\/18531"}],"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=12937"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=12937"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=12937"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}