{"id":59369,"date":"2025-10-25T05:10:27","date_gmt":"2025-10-25T05:10:27","guid":{"rendered":"http:\/\/zydq.1006ss.com\/?p=59369"},"modified":"2025-10-25T05:10:27","modified_gmt":"2025-10-25T05:10:27","slug":"%e3%80%90%e5%9f%ba%e7%a1%80%e7%a0%94%e7%a9%b6%e3%80%91%e8%90%9d%e5%8d%9c%e7%a1%ab%e7%b4%a0%e9%80%9a%e8%bf%87%e4%bf%83%e8%bf%9b%e5%89%8d%e9%a2%9d%e5%8f%91%e8%82%b2%e5%92%8c%e7%a5%9e%e7%bb%8f%e5%85%83","status":"publish","type":"post","link":"http:\/\/zydq.1006ss.com\/?p=59369","title":{"rendered":"\u3010\u57fa\u7840\u7814\u7a76\u3011\u841d\u535c\u786b\u7d20\u901a\u8fc7\u4fc3\u8fdb\u524d\u989d\u53d1\u80b2\u548c\u795e\u7ecf\u5143\u5206\u5316\u4fc3\u8fdb\u5927\u8111\u7c7b\u5668\u5b98\u751f\u957f"},"content":{"rendered":"

\u6587\u7ae0\u6765\u6e90\uff1a\u4e2d\u534e\u7cbe\u795e\u79d1\u6742\u5fd7, 2022,55(3) : 204-213<\/p>\n

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\u6458\u8981<\/p>\n

\u76ee\u7684<\/p>\n

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\u65b9\u6cd5<\/p>\n

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\u7ed3\u679c<\/p>\n

\u52a0\u5165\u841d\u535c\u786b\u7d20\u5904\u740640 d\u7684\u5927\u8111\u7c7b\u5668\u5b98\u4e2d\u6027\u522b\u51b3\u5b9a\u533aY\u6846\u86cb\u767d2\uff08sex determining region Y-box 2\uff0cSOX2\uff09\u3001\u524d\u8111\u6807\u5fd7\u86cb\u767d\u53c9\u5934\u6846G1\uff08forkhead box G1\uff0cFOXG1\uff09\u3001\u524d\u989d\u53f6\u76ae\u8d28\u5b64\u72ec\u75c7\u6613\u611f\u5019\u9009\u57fa\u56e02\uff08autism susceptibility candidate 2\uff0cAUST2\uff09\u3001\u914d\u5bf9\u76d2\u57fa\u56e06\uff08paired box 6\uff0cPAX6\uff09\u8868\u8fbe\u660e\u663e\u589e\u5f3a\uff1b\u52a0\u5165\u841d\u535c\u786b\u7d20\u5904\u740670 d\u5927\u8111\u7c7b\u5668\u5b98\u4e2d\u795e\u7ecf\u5143\u6807\u5fd7\u86cb\u767d\u795e\u7ecf\u5143\u2162\u578b\u03b2-\u5fae\u7ba1\u86cb\u767d\uff08\u03b2-tubulin\uff0cTUJ1\uff09\u3001\u795e\u7ecf\u5143\u6838\uff08neuronal nuclei\uff0cNeuN\uff09\u3001\u5fae\u7ba1\u76f8\u5173\u86cb\u767d2\uff08recombinant microtubule associated protein 2\uff0cMAP2\uff09\u3001T\u8111\u86cb\u767d\u5bb6\u65cf1\uff08T-brain-1\uff0cTBR1\uff09\u7b49\u8868\u8fbe\u589e\u5f3a\u3002RNA\u6d4b\u5e8f\u5206\u6790\u663e\u793a\uff0c\u76f8\u6bd4\u4e8e\u5bf9\u7167\u7ec4\uff0c\u5b9e\u9a8c\u7ec4\u663e\u8457\u4e0a\u8c03\u57fa\u56e0105\u4e2a\uff0c\u4e0b\u8c03\u57fa\u56e0512\u4e2a\uff0c\u5171617\u4e2a\u3002qRT-PCR\u9a8c\u8bc1\u5dee\u5f02\u8868\u8fbe\u57fa\u56e0\uff08SFTPC\u3001AKR1C3\u3001CXCR6\u3001PRAP1\u3001TMC8\u3001GPR182\u3001F2RL2\u3001KCNJ10\uff09\u7684\u8f6c\u5f55\u6c34\u5e73\u4e0e\u6d4b\u5e8f\u7ed3\u679c\u57fa\u672c\u4e00\u81f4\u3002<\/p>\n

\u7ed3\u8bba<\/p>\n

\u5dee\u5f02\u8868\u8fbe\u57fa\u56e0AKR1C3\u3001KCNJ10\u53ef\u80fd\u53c2\u4e0e\u841d\u535c\u786b\u7d20\u4fc3\u8fdb\u5927\u8111\u7c7b\u5668\u5b98\u524d\u989d\u53d1\u80b2\u548c\u795e\u7ecf\u5143\u5206\u5316\uff0c\u8fd9\u6709\u52a9\u4e8e\u4e3a\u841d\u535c\u786b\u7d20\u6539\u5584\u7cbe\u795e\u75be\u75c5\u548c\u795e\u7ecf\u9000\u884c\u6027\u75be\u75c5\u60a3\u8005\u7684\u8ba4\u77e5\u548c\u75c7\u72b6\u63d0\u4f9b\u8bc1\u636e\u652f\u6301\u3002<\/p>\n

\u841d\u535c\u786b\u7d20\uff08sulforaphane\uff09\u662f\u4e00\u79cd\u5f02\u786b\u6c30\u9178\u76d0\uff0c\u7531\u786b\u4ee3\u8461\u8404\u7cd6\u82f7\uff08glucosinlate\uff09\u7ecf\u690d\u7269\u4f53\u5185\u9ed1\u82a5\u5b50\u9176\uff08myrosinase enzyme\uff09\u6c34\u89e3\u6240\u5f97\uff3b<\/sup>1\uff3d<\/sup>\u3002\u4f5c\u4e3a\u4e00\u79cd\u5e38\u89c1\u7684\u6297\u6c27\u5316\u5242\uff0c\u841d\u535c\u786b\u7d20\u5728\u82b1\u6930\u83dc\u3001\u7518\u84dd\u3001\u7fbd\u8863\u7518\u84dd\u548c\u82bd\u7518\u84dd\u3001\u6c34\u82b9\u7b49\u5341\u5b57\u82b1\u79d1\u852c\u83dc\u4e2d\u542b\u91cf\u8f83\u4e3a\u4e30\u5bcc\uff0c\u662f\u852c\u83dc\u4e2d\u6240\u53d1\u73b0\u7684\u6297\u764c\u6548\u679c\u6700\u597d\u7684\u690d\u7269\u6d3b\u6027\u7269\u8d28\u4e4b\u4e00\uff3b<\/sup>2, 3\uff3d<\/sup>\u3002\u81ea1948\u5e74\u53d1\u73b0\u4ee5\u6765\uff0c\u8fd9\u79cd\u786b\u4ee3\u8461\u8404\u7cd6\u82f7\u88ab\u9891\u7e41\u7814\u7a76\uff0c\u5e76\u663e\u793a\u51fa\u591a\u79cd\u751f\u7269\u6548\u5e94\uff0c\u5305\u62ec\u6291\u5236\u6c27\u5316\u5e94\u6fc0\u3001\u6297\u708e\u3001\u6297\u83cc\u3001\u6297\u764c\u3001\u6297\u8870\u8001\u3001\u795e\u7ecf\u4fdd\u62a4\u548c\u6297\u7cd6\u5c3f\u75c5\uff3b<\/sup>4, 5\uff3d<\/sup>\u3002\u8fd1\u5e74\u6765\u7814\u7a76\u663e\u793a\uff0c\u841d\u535c\u786b\u7d20\u80fd\u591f\u6539\u5584\u5927\u9f20\u53ca\u5c0f\u9f20\u5b66\u4e60\u8bb0\u5fc6\u529f\u80fd\u969c\u788d\uff0c\u5728\u963f\u5c14\u8328\u6d77\u9ed8\u75c5\uff08Alzheimer\u2032s disease\uff0cAD\uff09\u3001\u5e15\u91d1\u68ee\u75c5\u3001\u6291\u90c1\u75c7\u3001\u7cbe\u795e\u5206\u88c2\u75c7\u3001\u81ea\u95ed\u75c7\u7b49\u75be\u75c5\u7684\u8ba4\u77e5\u529f\u80fd\u6539\u5584\u4e2d\u8d77\u91cd\u8981\u4f5c\u7528\uff3b<\/sup>6, 7, 8\uff3d<\/sup>\u3002<\/p>\n

\u5927\u8111\u7c7b\u5668\u5b98\u662f\u7531\u4eba\u8bf1\u5bfc\u591a\u80fd\u5e72\u7ec6\u80de\uff08human induced pluripotent stem cells\uff0cHiPSCs\uff09\u5b9a\u5411\u8bf1\u5bfc\u5206\u5316\u5f62\u6210\u7684\u7c7b\u4f3c\u5927\u8111\u65e9\u671f\u53d1\u80b2\u8fc7\u7a0b\u4e2d\u7684\u76ae\u8d28\u7ed3\u6784\uff3b<\/sup>9, 10\uff3d<\/sup>\u3002\u5176\u7531Lancaster\u7b49\uff3b<\/sup>11\uff3d<\/sup>\u57282013\u5e74\u5efa\u7acb\uff0c\u80fd\u8f83\u597d\u5730\u53cd\u6620\u4eba\u4f53\u65e9\u671f\u80da\u80ce\u671f\u5927\u8111\u7684\u53d1\u80b2\u8fc7\u7a0b\u3002RNA\u6d4b\u5e8f\u663e\u793a\uff0c\u57f9\u517b\u81f3\u7b2c40\u5929\u7684\u5927\u8111\u7c7b\u5668\u5b98\u4e0e\u5b559\u5468\u7684\u4eba\u7c7b\u80ce\u513f\u5927\u8111\u6807\u672c\u7684\u57fa\u56e0\u8868\u8fbe\u8c31\u5341\u5206\u76f8\u4f3c\uff0c\u57f9\u517b70 d\u7684\u5927\u8111\u7c7b\u5668\u5b98\u4e0e\u5b55\u4e2d\u671f\u7684\u5927\u8111\u57fa\u56e0\u8868\u8fbe\u8c31\u7c7b\u4f3c\uff0c\u63d0\u793a\u5927\u8111\u7c7b\u5668\u5b98\u80fd\u591f\u5728\u4e00\u5b9a\u7a0b\u5ea6\u4e0a\u53cd\u6620\u4f53\u5185\u65e9\u671f\u80da\u80ce\u5927\u8111\u7684\u5f62\u6210\uff3b<\/sup>12\uff3d<\/sup>\u3002\u56e0\u6b64\uff0c\u5927\u8111\u7c7b\u5668\u5b98\u53ef\u4ee5\u4f5c\u4e3a\u4e00\u79cd\u6709\u6548\u7684\u4f53\u5916\u65e9\u671f\u5927\u8111\u53d1\u80b2\u6a21\u578b\uff0c\u7528\u4e8e\u7814\u7a76\u841d\u535c\u786b\u7d20\u5bf9\u5927\u8111\u53d1\u80b2\u7684\u5f71\u54cd\u3002\u4e0e\u5e38\u7528\u7684\u795e\u7ecf\u7ec6\u80de\u57f9\u517b\u548c\u52a8\u7269\u6a21\u578b\u76f8\u6bd4\uff0c\u5927\u8111\u7c7b\u5668\u5b98\u53ef\u4ee5\u5728\u4e00\u5b9a\u7a0b\u5ea6\u4e0a\u4f53\u5916\u6a21\u62df\u4eba\u7c7b\u5927\u8111\u7684\u53d1\u80b2\u8fc7\u7a0b\u548c\u7ed3\u6784\u7279\u70b9\uff0c\u5e76\u80fd\u8f83\u597d\u5730\u4fdd\u6301\u4eba\u4f53\u7279\u5f02\u6027\u7684\u57fa\u56e0\u578b\u548c\u86cb\u767d\u8868\u8fbe\u6c34\u5e73\uff0c\u76ee\u524d\u5e7f\u6cdb\u7528\u4e8e\u6a21\u62df\u795e\u7ecf\u53d1\u80b2\u3001\u611f\u67d3\u6027\u548c\u795e\u7ecf\u7cbe\u795e\u75be\u75c5\u7684\u75c5\u56e0\u5b66\u548c\u75c5\u7406\u8fc7\u7a0b\u4e2d\uff3b<\/sup>13, 14, 15, 16\uff3d<\/sup>\u3002<\/p>\n

\u6211\u4eec\u4f7f\u7528\u841d\u535c\u786b\u7d20\u5904\u7406HiPSCs\u5206\u5316\u5f62\u6210\u7684\u5927\u8111\u7c7b\u5668\u5b98\uff0c\u901a\u8fc7\u6bd4\u8f83\u4e0d\u540c\u65f6\u671f\u5927\u8111\u7c7b\u5668\u5b98\u8111\u533a\u5206\u5316\u548c\u795e\u7ecf\u7ec6\u80de\u6807\u5fd7\u7269\u7684\u6539\u53d8\uff0c\u63a2\u8ba8\u841d\u535c\u786b\u7d20\u5bf9\u5927\u8111\u7c7b\u5668\u5b98\u53d1\u80b2\u7684\u5f71\u54cd\uff1b\u5e76\u8fdb\u4e00\u6b65\u8fdb\u884c\u8f6c\u5f55\u7ec4\u5b66\u7814\u7a76\u53ca\u76f8\u5173\u751f\u7269\u4fe1\u606f\u5b66\u5206\u6790\uff0c\u8fdb\u4e00\u6b65\u63a2\u7d22\u841d\u535c\u786b\u7d20\u5f71\u54cd\u5927\u8111\u7c7b\u5668\u5b98\u7684\u53ef\u80fd\u673a\u5236\u3002<\/p>\n

\u6750\u6599\u548c\u65b9\u6cd5<\/p>\n

\u4e00\u3001\u6750\u6599<\/p>\n

\u672c\u7814\u7a76\u4e3a\u5e72\u9884\u6027\u7814\u7a76\uff0c\u7814\u7a76\u8d77\u6b62\u65f6\u95f4\u4e3a2020\u5e7410\u6708\u81f32021\u5e7410\u6708\u3002<\/p>\n

\u5b9e\u9a8c\u7ec6\u80de\u548c\u4e3b\u8981\u8bd5\u5242\uff1aHiPSCs\u7ec6\u80de\u7cfb\u8d2d\u81ea\u5317\u4eac\u8d5b\u8d1d\u751f\u7269\u6280\u672f\u6709\u9650\u516c\u53f8\uff08\u8d27\u53f7CA4024106\u3001CA4025106\u53caCA4028106\uff09\uff0cMatrigel\u5de5\u4f5c\u6db2\uff08\u7f8e\u56fdCorning\u516c\u53f8\uff09\uff0cHiPSCs\u7ec6\u80de\u57f9\u517b\u4f7f\u7528mTeSR\u21221\u57f9\u517b\u57fa\uff08\u7f8e\u56fdSTEM CELL\u516c\u53f8\uff09\uff0c\u5927\u8111\u7c7b\u5668\u5b98\u57f9\u517b\u4f7f\u7528STEMdiff\u2122 Cerebral Organoid Kit\u57f9\u517b\u57fa\uff08\u7f8e\u56fdSTEM CELL\u516c\u53f8\uff09\uff0c\u841d\u535c\u786b\u7d20\uff08\u7f8e\u56fdSelleck\u516c\u53f8\uff09\uff0c\u6297\u4f53\uff1a\u516b\u805a\u4f53\u8f6c\u5f55\u56e0\u5b504\uff08ecombinant octamer binding transcription factor 4\uff0cOCT4\uff1b\u7f8e\u56fdGENE TEX\u516c\u53f8\uff09\u548c\u9636\u6bb5\u7279\u5f02\u6027\u80da\u80ce\u6297\u539f4\uff08stage-specific embryonic antigen 4\uff0cSSEA4\uff1b\u7f8e\u56fdGENE TEX\u516c\u53f8\uff09\uff0cKi-67\uff08\u82f1\u56fdAbcam\u516c\u53f8\uff09\u548c\u534a\u80f1\u6c28\u9178\u7684\u5929\u51ac\u6c28\u9178\u86cb\u767d\u6c34\u89e3\u9176-3\uff08Caspas-3\uff1b\u82f1\u56fdAbcam\u516c\u53f8\uff09\u3001\u524d\u8111\u6807\u5fd7\u86cb\u767d\u53c9\u5934\u6846G1\uff08forkhead box G1\uff0cFOXG1\uff1b\u82f1\u56fdAbcam\u516c\u53f8\uff09\u3001\u6d77\u9a6c\u7684\u6807\u5fd7\u86cb\u767d\u5377\u66f2\u86cb\u767d9\uff08Frizzled 9\uff0c\u82f1\u56fdAbcam\u516c\u53f8\uff09\u3001\u795e\u7ecf\u5e72\u7ec6\u80de\u6807\u5fd7\u86cb\u767d\u6027\u522b\u51b3\u5b9a\u533aY\u6846\u86cb\u767d2\uff08sex determining region Y-box 2\uff0cSOX2\uff1b\u7f8e\u56fdCell Signaling\u516c\u53f8\uff09\u3001\u7c7b\u8111\u5ba4\u7ed3\u6784\u6807\u5fd7\u86cb\u767dN-\u9499\u9ecf\u86cb\u767d\uff08N-cadherin\uff1b\u7f8e\u56fdCell signaling\u516c\u53f8\uff09\u3001\u524d\u989d\u76ae\u8d28\u6807\u5fd7\u86cb\u767d\u5b64\u72ec\u75c7\u6613\u611f\u5019\u9009\u57fa\u56e02\uff08autism susceptibility candidate 2\uff0cAUTS2\uff1b\u7f8e\u56fdProteintech\u516c\u53f8\uff09\u3001\u795e\u7ecf\u5143\u548c\u76ae\u8d28\u53d1\u80b2\u6807\u5fd7\u86cb\u767d\u914d\u5bf9\u76d2\u57fa\u56e06\uff08paired box 6\uff0cPAX6\uff1b\u82f1\u56fdAbcam\u516c\u53f8\uff09\u3001\u6df1\u5c42\u76ae\u8d28\u6807\u5fd7\u7269T\u8111\u86cb\u767d\u5bb6\u65cf1\uff08T-brain-1\uff0cTBR1\uff1b\u82f1\u56fdAbcam\u516c\u53f8\uff09\uff0c\u5c11\u7a81\u80f6\u8d28\u7ec6\u80de\u7cfb\u8f6c\u5f55\u56e0\u5b502\uff08recombinant oligodendrocyte lineage\uff0cOLIG2\uff1b\u82f1\u56fdAbcam\u516c\u53f8\uff09\u548c\u795e\u7ecf\u7ec6\u80de\u6807\u5fd7\u86cb\u767d\u795e\u7ecf\u5143\u2162\u578b\u03b2-\u5fae\u7ba1\u86cb\u767d\uff08\u03b2-tubulin\uff0cTUJ1\uff1b\u7f8e\u56fdcell signaling\u516c\u53f8\uff09\u3001\u5fae\u7ba1\u76f8\u5173\u86cb\u767d2\uff08recombinant microtubule associated protein 2\uff0cMAP2\uff1b\u82f1\u56fdAbcam\u516c\u53f8\uff09\u3001\u795e\u7ecf\u5143\u6838\uff08neuronal nuclei\uff0cNeuN\uff1b\u82f1\u56fdAbcam\u516c\u53f8\uff09\u53ca\u80f6\u8d28\u7ea4\u7ef4\u9178\u6027\u86cb\u767d\uff08glial fibrillary acidic protein\uff0cGFAP\uff1b\u7f8e\u56fdCell Signaling\u516c\u53f8\uff09\u7b49\u3002\u4e8c\u6297\uff1a\u5c71\u7f8a\u6297\u5154IgG\uff08H+L\uff09\u9ad8\u4ea4\u53c9\u5438\u9644\u4e8c\u6297\u3001\u5c71\u7f8a\u6297\u9f20IgG\uff08H+L\uff09\u4ea4\u53c9\u5438\u9644\u4e8c\u6297\uff0c\u5c71\u7f8a\u6297\u5154IgG\uff08H+L\uff09\u4ea4\u53c9\u5438\u9644\u4e8c\u6297\u548c\u5c71\u7f8a\u6297\u5c0f\u9f20IgG\uff08H+L\uff09\u4ea4\u53c9\u5438\u9644\u4e8c\u6297\uff08\u5747\u4e3a\u7f8e\u56fdInvitrogen\u516c\u53f8\uff09\uff1b\u53cd\u8f6c\u5f55\u8bd5\u5242\u76d2\uff08\u6e56\u5357\u827e\u79d1\u745e\u751f\u7269\u5de5\u7a0b\u6709\u9650\u516c\u53f8\uff0cAG11711\uff09\u3002<\/p>\n

\u91c7\u7528\u5b8c\u5168\u968f\u673a\u5316\u8fdb\u884c\u5206\u7ec4\uff0c\u5b9e\u9a8c\u7ec4\u4e3a\u4e0d\u540c\u6d53\u5ea6\uff081 \u03bcmol\/L\u30012 \u03bcmol\/L\u30015 \u03bcmol\/L\uff09\u841d\u535c\u786b\u7d20\u5904\u7406\u7ec4\uff0c\u5bf9\u7167\u7ec4\u4e3a\u7a7a\u767d\u5bf9\u7167\u7ec4\uff0c\u6bcf\u7ec420\u4e2a\u7c7b\u5668\u5b98\u3002<\/p>\n

\u672c\u7814\u7a76\u65b9\u6848\u7ecf\u8fc7\u4e2d\u5357\u5927\u5b66\u6e58\u96c5\u4e8c\u533b\u9662\u4f26\u7406\u59d4\u5458\u4f1a\u6279\u51c6\uff0c\u6279\u53f7\u4e3a\uff1a2021723\u3002<\/p>\n

\u4e8c\u3001\u65b9\u6cd5<\/p>\n

1.\u5927\u8111\u7c7b\u5668\u5b98\u7684\u57f9\u517b\uff1a\u5728\u57f9\u517bHiPSCs\u7ec6\u80de\u65f6\uff0c\u63d0\u524d\u5229\u7528Matrigel\u5de5\u4f5c\u6db2\u8fdb\u884c\u7ec6\u80de\u57f9\u517b\u76bf\u7684\u94fa\u5e95\u9884\u5904\u7406\uff0c\u4f7f\u7528mTeSR\u21221\u8fdb\u884cHiPSCs\u7ec6\u80de\u57f9\u517b\u3002\u5c06mTeSR\u21221\u57f9\u517b\u57fa\u4e2d\u7684HiPSCs\u6dfb\u52a0\u5230\u4f4e\u7c98\u964496\u5b54\u677f\u7684\u6bcf\u4e2a\u5b54\u4e2d\uff0c\u6bcf\u4e2a\u5b54\u52a0\u5165STEMdiff\u2122 Cerebral Organoid Kit\u57f9\u517b\u57fa\uff0c\u653e\u516537 \u2103\u7684\u57f9\u517b\u7bb1\uff085%CO2<\/sub>\uff0c21%O2<\/sub>\uff09\u4e2d\u57f9\u517b\uff0c\u5f62\u6210\u62df\u80da\u4f53\u3002\u7b2c5\u5929\uff0c\u5c06\u5f62\u6210\u7684\u62df\u80da\u4f53\u8f6c\u79fb\u5230\u4f4e\u7c98\u964424\u5b54\u677f\u4e2d\u5e76\u57f9\u517b2 d\uff0c\u5f62\u6210\u4e2d\u5fc3\u8f83\u9ed1\uff0c\u8fb9\u7f18\u6e05\u6670\u5149\u6ed1\u7684\u795e\u7ecf\u5916\u80da\u5c42\u3002\u7b2c8\u5929\uff0c\u5c06\u7ec4\u7ec7\u5305\u57cb\u5728Matrigel\u80f6\u4e2d\u5e76\u8f6c\u79fb\u81f3\u4f4e\u7c98\u96446\u5b54\u677f\u4e2d\u3002\u7b2c10\u5929\uff0c\u7ec4\u7ec7\u5468\u56f4\u795e\u7ecf\u4e0a\u76ae\u82bd\u7a81\u51fa\u73b0\uff0c\u5c06\u57f9\u517b\u76bf\u8f6c\u79fb\u81f3\u57f9\u517b\u7bb1\u4e2d\u7684\u6447\u5e8a\u4e0a\uff0c\u9707\u8361\u57f9\u517b\uff0c\u540c\u65f6\u66f4\u6362\u6210\u7c7b\u5668\u5b98\u6210\u719f\u57f9\u517b\u57fa\u8fdb\u884c\u957f\u671f\u57f9\u517b\u3002\u4e8e\u7b2c8\u5929\u5c06\u4e0d\u540c\u6d53\u5ea6\uff081 \u03bcmol\/L\u30012 \u03bcmol\/L\u30015 \u03bcmol\/L\uff09\u7684\u841d\u535c\u786b\u7d20\u6dfb\u52a0\u81f3\u57f9\u517b\u76bf\u4e2d\uff0c\u5206\u522b\u57f9\u517b\u81f340 d\u300170 d\u3002\u6bcf\u5929\u5728\u663e\u5fae\u955c\u4e0b\u68c0\u67e5HiPSCs\u3001\u62df\u80da\u4f53\u3001\u795e\u7ecf\u5916\u80da\u5c42\u3001\u795e\u7ecf\u4e0a\u76ae\u7ec4\u7ec7\u548c\u5927\u8111\u7c7b\u5668\u5b98\u7684\u5f62\u6001\u548c\u751f\u957f\u60c5\u51b5\uff0c\u5e76\u901a\u8fc7\u8367\u5149\u663e\u5fae\u955c\uff08\u65e5\u672cOLYMPUS\u516c\u53f8\uff0c\u578b\u53f7BX51\uff09\u5728\u660e\u4eae\u89c6\u91ce\u4e0b\u6210\u50cf\u3002<\/p>\n

2. \u82cf\u6728\u7cbe-\u4f0a\u7ea2\u67d3\u8272\uff08hematoxylin and eosin staining\uff0cHE\uff09\u4e0e\u514d\u75ab\u8367\u5149\u67d3\u8272\uff1a\u5927\u8111\u7c7b\u5668\u5b98\u75284%\u591a\u805a\u7532\u919b\u56fa\u5b9a\u3001\u8131\u6c34\u3001\u900f\u660e\u3001\u77f3\u8721\u6d78\u6ce1\u548c\u77f3\u8721\u5305\u57cb\u540e\u8fdb\u884c\u4e0b\u4e00\u6b65\u5207\u7247\u64cd\u4f5c\u3002\u67d3\u8272\u65f6\uff0c\u9996\u5148\u5c06\u5207\u7247\u7f6e\u4e8e60 \u2103\u70d8\u7bb1\u4e2d1 h\u7194\u5316\u77f3\u8721\uff0c\u968f\u540e\u7f6e\u4e8e\u4e8c\u7532\u82ef\u7684\u6eb6\u6db2\u4e2d\u6d78\u6ce1\uff0c\u7136\u540e\u7528\u4e0d\u540c\u68af\u5ea6\u9152\u7cbe\u5904\u7406\u540e\u5728\u5fae\u6ce2\u7089\u4e2d\u8fdb\u884c\u6297\u539f\u4fee\u590d\uff08\u67e0\u6aac\u9178\u94a0\uff0cpH=6.0\uff09\u3002\u6ef4\u52a0\u5c71\u7f8a\u8840\u6e05\u5c01\u95ed\u6db2\uff0c\u5ba4\u6e29\u5c01\u95ed1 h\u540e\u52a0\u4e00\u62974 \u2103\u8fc7\u591c\u5b75\u80b2\u3002\u7b2c2\u5929\u5728\u6807\u672c\u4e0a\u6ef4\u52a0\u8367\u5149\u7d20\u6807\u8bb0\u7684\u4e8c\u6297\uff0c\u5ba4\u6e29\u5b75\u80b230 min\u81f31 h\uff0c\u78f7\u9178\u7f13\u51b2\u76d0\u6eb6\u6db2\uff08phosphate buffered saline\uff0cPBS\uff09\u6e05\u6d17\u540e\u75284\u2032\uff0c6-\u4e8c\u8112\u57fa-2-\u82ef\u57fa\u5432\u54da\uff084\u2032\uff0c6-Diamidino-2-Phenylindole\uff0cDAPI\uff09\u67d3\u827210 min\uff0c\u7528\u9632\u6dec\u706d\u8367\u5149\u5242\u5c01\u7247\u3002<\/p>\n

HE\u67d3\u8272\uff1a\u77f3\u8721\u5207\u7247\u7ecf\u8131\u8721\u3001\u68af\u5ea6\u9152\u7cbe\u8131\u6c34\u540e\uff0c\u4f7f\u7528\u82cf\u6728\u7cbe\u67d3\u6599\u8fdb\u884c\u67d3\u8272\uff0c\u6d41\u6c34\u7a0d\u6d17\u53bb\u67d3\u6599\uff0c\u518d\u75280.1%\u76d0\u9178\u4e59\u9187\u8fdb\u884c\u5206\u5316\uff0c\u7136\u540e\u7528\u4f0a\u7ea2\u6db2\u67d3\u8272\uff0c\u67d3\u8272\u5b8c\u6210\u540e\u7528\u6d41\u6c34\u6d17\u53bb\u591a\u4f59\u67d3\u6599\uff0c\u6c34\u6d17\u5b8c\u67d3\u6599\u540e\u7528\u4ece\u4f4e\u5230\u9ad8\u6d53\u5ea6\u7684\u9152\u7cbe\u8fdb\u884c\u9187\u6d17\u4ee5\u53ca\u8131\u6c34\uff0c\u4f7f\u7528\u4e8c\u7532\u82ef\u8fdb\u884c\u900f\u660e\uff0c\u6811\u8102\u8fdb\u884c\u5c01\u7247\u3002<\/p>\n

3.RNA\u6d4b\u5e8f\uff1a\u5bf9\u4f7f\u75281 \u03bcmol\/L\u841d\u535c\u786b\u7d20\u5904\u7406\u540e\u7b2c40\u5929\u7684\u5b9e\u9a8c\u7ec4\u548c\u5bf9\u7167\u7ec4\u5927\u8111\u7c7b\u5668\u5b98\u540c\u65f6\u8fdb\u884cRNA\u6d4b\u5e8f\uff0c\u6bcf\u7ec43\u4e2a\u6837\u54c1\u3002\u5229\u7528\u771f\u6838\u751f\u7269\u5927\u90e8\u5206\u4fe1\u4f7fRNA\uff08messenger RNA\uff0cmRNA\uff09\u90fd\u5e26\u6709\u591a\u805a\u6838\u82f7\u9178\uff08polynucleotides\uff0cPolyA\uff09\u5c3e\u7684\u7ed3\u6784\u7279\u5f81\uff0c\u901a\u8fc7Oligo\uff08dT\uff09\u78c1\u73e0\uff08\u7f8e\u56fdThermo\u516c\u53f8\uff09\u5bcc\u96c6\u5e26\u6709Poly A\u5c3e\u7684mRNA\uff0c\u968f\u540e\u5728\u9650\u5236\u6027\u5185\u5207\u9176\uff08NEB\uff09RNA\u6587\u5e93\u51c6\u5907\u5de5\u5177\u5305\u4e2d\u7528\u4e8c\u4ef7\u9633\u79bb\u5b50\u5c06\u5f97\u5230\u7684mRNA\u968f\u673a\u6253\u65ad\uff0c\u6309\u7167NEB\u666e\u901a\u5efa\u5e93\u65b9\u5f0f\u6216\u94fe\u7279\u5f02\u6027\u5efa\u5e93\u65b9\u5f0f\u8fdb\u884c\u5efa\u5e93\u3002\u6d4b\u5e8f\u7684\u6838\u5fc3\u662f\u57fa\u56e0\u8868\u8fbe\u5dee\u5f02\u7684\u663e\u8457\u6027\u5206\u6790\uff0c\u91c7\u7528\u7edf\u8ba1\u5b66\u65b9\u6cd5\u6bd4\u8f832\u4e2a\u6761\u4ef6\u6216\u591a\u4e2a\u6761\u4ef6\u4e0b\u7684\u57fa\u56e0\u8868\u8fbe\u5dee\u5f02\uff0c\u4ece\u4e2d\u627e\u51fa\u4e0e\u6761\u4ef6\u76f8\u5173\u7684\u7279\u5f02\u6027\u57fa\u56e0\uff0c\u7136\u540e\u8fdb\u4e00\u6b65\u4f7f\u7528IPA\u8f6f\u4ef6\uff08Qiagen\u516c\u53f8\uff09\u5bf9\u6bcf\u7ec4\u7684mRNA\u4e30\u5ea6\u8fdb\u884c\u751f\u7269\u4fe1\u606f\u5b66\u8bc4\u4f30\u3002\u5dee\u5f02\u57fa\u56e0\u7684\u4eac\u90fd\u57fa\u56e0\u4e0e\u57fa\u56e0\u7ec4\u767e\u79d1\u5168\u4e66\uff08Kyoto Encyclopedia of Genes and Genomes\uff0cKEGG\uff09\u5206\u6790\u548c\u57fa\u56e0\u672c\u4f53\u8bba\uff08Gene Ontology\uff0cGO\uff09\u5206\u6790\uff1aKEGG\u662f\u6574\u5408\u4e86\u57fa\u56e0\u7ec4\u3001\u5316\u5b66\u548c\u7cfb\u7edf\u529f\u80fd\u4fe1\u606f\u7684\u7efc\u5408\u6027\u6570\u636e\u5e93\u3002KEGG\u901a\u8def\u5bcc\u96c6\u4ee5Padj<\/sub><0.05\u4f5c\u4e3a\u663e\u8457\u6027\u5bcc\u96c6\u7684\u9608\u503c\u3002GO\u662f\u63cf\u8ff0\u57fa\u56e0\u529f\u80fd\u7684\u7efc\u5408\u6027\u6570\u636e\u5e93\uff0c\u53ef\u5206\u4e3a\u751f\u7269\u8fc7\u7a0b\uff08biological process\uff09\u548c\u7ec6\u80de\u7ec4\u6210\uff08cellular component\uff09\u3001\u5206\u5b50\u529f\u80fd\uff08molecular function\uff093\u4e2a\u90e8\u5206\u3002GO\u529f\u80fd\u5bcc\u96c6\u4ee5Padj<\/sub><0.05\u4f5c\u4e3a\u663e\u8457\u6027\u5bcc\u96c6\u7684\u9608\u503c\u3002\u57fa\u56e0\u5bcc\u96c6\u5206\u6790\uff08gene set enrichment analysis\uff0cGSEA\uff09\uff1a\u662f\u4e00\u79cd\u9488\u5bf9\u5168\u57fa\u56e0\u7ec4\u8868\u8fbe\u8c31\u82af\u7247\u6570\u636e\u7684\u5206\u6790\u65b9\u6cd5\uff0c\u5c06\u57fa\u56e0\u4e0e\u9884\u5b9a\u4e49\u7684\u57fa\u56e0\u96c6\u8fdb\u884c\u6bd4\u8f83\u3002\u5373\u7efc\u5408\u73b0\u6709\u7684\u5bf9\u57fa\u56e0\u7684\u5b9a\u4f4d\u3001\u6027\u8d28\u3001\u529f\u80fd\u3001\u751f\u7269\u5b66\u610f\u4e49\u7b49\u4fe1\u606f\u57fa\u7840\uff0c\u6784\u5efa\u4e00\u4e2a\u5206\u5b50\u6807\u7b7e\u6570\u636e\u5e93\uff0c\u5728\u6b64\u6570\u636e\u5e93\u4e2d\u5c06\u5df2\u77e5\u57fa\u56e0\u6309\u7167\u67d3\u8272\u4f53\u4f4d\u7f6e\u3001\u5df2\u5efa\u7acb\u57fa\u56e0\u96c6\u3001\u6a21\u5e8f\u3001\u80bf\u7624\u76f8\u5173\u57fa\u56e0\u96c6\u548cGO\u57fa\u56e0\u96c6\u7b49\u591a\u4e2a\u529f\u80fd\u57fa\u56e0\u96c6\u8fdb\u884c\u5206\u7ec4\u4e0e\u5f52\u7c7b\u3002\u901a\u8fc7\u5206\u6790\u57fa\u56e0\u8868\u8fbe\u8c31\u6570\u636e\uff0c\u4e86\u89e3\u5176\u5728\u7279\u5b9a\u7684\u529f\u80fd\u57fa\u56e0\u96c6\u4e2d\u7684\u8868\u8fbe\u72b6\u51b5\uff0c\u4ee5\u53ca\u8fd9\u79cd\u8868\u8fbe\u72b6\u51b5\u662f\u5426\u5b58\u5728\u7edf\u8ba1\u5b66\u610f\u4e49\u3002GSEA\u4e0d\u9700\u8981\u6307\u5b9a\u660e\u786e\u7684\u5dee\u5f02\u57fa\u56e0\u9608\u503c\uff0c\u628a\u57fa\u56e0\u6309\u7167\u57282\u7ec4\u6837\u672c\u4e2d\u7684\u5dee\u5f02\u8868\u8fbe\u7a0b\u5ea6\u8fdb\u884c\u6392\u5e8f\uff0c\u7136\u540e\u91c7\u7528\u7edf\u8ba1\u5b66\u65b9\u6cd5\u68c0\u9a8c\u9884\u5148\u8bbe\u5b9a\u7684\u57fa\u56e0\u96c6\u5408\u662f\u5426\u5728\u6392\u5e8f\u8868\u7684\u9876\u7aef\u6216\u4f4e\u6bb5\u5bcc\u96c6\u3002GSEA\u4e3b\u8981\u5305\u62ec3\u4e2a\u6b65\u9aa4\uff1a\u8ba1\u7b97\u5bcc\u96c6\u5f97\u5206\uff08enrichment score\uff09\uff0c\u4f30\u8ba1\u5bcc\u96c6\u5f97\u5206\u7684\u663e\u8457\u6027\u6c34\u5e73\uff0c\u591a\u91cd\u5047\u8bbe\u68c0\u9a8c\u3002<\/p>\n

4.\u5b9e\u65f6\u5b9a\u91cf\u53cd\u8f6c\u5f55PCR\uff08real-time quantitative reverse transcriptase-mediated polymerase chain reaction\uff0cqRT-PCR\uff09\u9a8c\u8bc1\uff1a\u901a\u8fc7\u4e8c\u4ee3\u6d4b\u5e8f\u7ed3\u679c\uff0c\u9009\u53d6\u5b9e\u9a8c\u7ec4\u548c\u5bf9\u7167\u7ec4\u53d8\u5316\u500d\u65704\u500d\u4ee5\u4e0a\u4e14\u5177\u6709\u663e\u8457\u6027\u7684\u8fdb\u884cqRT-PCR\u7684\u9a8c\u8bc1\uff0c\u9009\u51fa\u6bd4\u8f83\u663e\u8457\u57fa\u56e0\u3002\u4f7f\u7528RNAfast200\u8bd5\u5242\u76d2\uff08\u4e0a\u6d77\u98de\u6377\u751f\u7269\u6280\u672f\u6709\u9650\u516c\u53f8\uff09\u63d0\u53d62\u4e2a\u4e0d\u540c\u7ec6\u80de\u682a\u841d\u535c\u786b\u7d20\u4e0e\u5bf9\u7167\u7ec4\u5927\u8111\u7c7b\u5668\u5b98\uff08\u6bcf\u4e2a\u6837\u54c13\u4e2a\u590d\u5b54\uff09\u7684\u603bRNA\uff0cNanoDrop2000\u5fae\u91cf\u5206\u5149\u5149\u5ea6\u8ba1\uff08\u7f8e\u56fdThermo\u516c\u53f8\uff09\u6d4b\u5b9aRNA\u6d53\u5ea6\uff0c\u4f7f\u7528\u53cd\u8f6c\u5f55\u8bd5\u5242\u76d2\u5c06RNA\u9006\u8f6c\u5f55\u5408\u6210cDNA\uff0c\u5728LightCycler 480\uff08\u7f57\u6c0f\u516c\u53f8\uff09\u901a\u8fc7\u5b9e\u65f6\u8367\u5149\u5b9a\u91cfPCR\u6cd5\u5bf9\u57fa\u56e0\u8868\u8fbe\u8fdb\u884c\u76f8\u5bf9\u5b9a\u91cf\u5206\u6790\u3002<\/p>\n

\u4e09\u3001\u7edf\u8ba1\u5b66\u5904\u7406<\/p>\n

\u514d\u75ab\u67d3\u8272\u56fe\u50cf\u4f7f\u7528Image Pro Plus 6.0\u8fdb\u884c\u7ec6\u80de\u8ba1\u6570\u3002\u5229\u7528GraphPad Prism 8\u8f6f\u4ef6\u8fdb\u884c\u514d\u75ab\u8367\u5149\u4e0d\u540c\u57fa\u56e0\u56fe\u50cf\u53caqRT-PCR\u57fa\u56e0\u8868\u8fbe\u91cf\u7edf\u8ba1\uff0c\u6570\u636e\u91c7\u7528\u5747\u6570\u00b1\u6807\u51c6\u5dee\u8868\u793a\uff0c\u91c7\u7528t\u68c0\u9a8c\u6bd4\u8f832\u7ec4\u514d\u75ab\u8367\u5149\u7ec6\u80de\u6570\u53caqRT-PCR\u57fa\u56e0\u8868\u8fbe\u91cf\uff0cP<0.05\u6216P<0.01\u4e3a\u5dee\u5f02\u6709\u7edf\u8ba1\u5b66\u610f\u4e49\u3002\u751f\u7269\u4fe1\u606f\u5b66\u5206\u6790\u91c7\u7528R\u8bed\u8a00\u8f6f\u4ef6\u7b5b\u9009\u5dee\u5f02\u57fa\u56e0\uff0c\u7136\u540e\u4f7f\u7528IPA\u8f6f\u4ef6\u8fdb\u884cGO\u3001KEGG\u53caGSEA\u5206\u6790\u3002<\/p>\n

\u7ed3\u679c<\/p>\n

\u4e00\u3001\u841d\u535c\u786b\u7d20\u4fc3\u8fdb\u5927\u8111\u7c7b\u5668\u5b98\u751f\u957f<\/p>\n

HiPSCs\u5728\u6b63\u5e38\u5e72\u7ec6\u80de\u57f9\u517b\u6761\u4ef6\u4e0b\u96c6\u843d\u6837\u751f\u957f\uff0c\u5e76\u8868\u8fbe\u591a\u80fd\u5e72\u7ec6\u80de\u6807\u8bb0OCT4\u3001SSEA4\u3001Ki-67\u548cCaspas-3\u3002\u5728\u5e72\u7ec6\u80de\u5b9a\u5411\u5206\u5316\u8fc7\u7a0b\u4e2d\uff0cHiPSCs\u9010\u6b65\u5206\u5316\u5f62\u6210\u62df\u80da\u4f53\u3001\u7b2c8\u5929\u5de6\u53f3\u5f62\u6210\u795e\u7ecf\u5916\u80da\u5c42\u3001\u7b2c10\u5929\u5de6\u53f3\u5f62\u6210\u795e\u7ecf\u4e0a\u76ae\u7ec4\u7ec7\uff0c\u957f\u671f\u57f9\u517b\u7684\u5927\u8111\u7c7b\u5668\u5b98\u5176\u7ec4\u7ec7\u5f62\u72b6\u8fc5\u901f\u589e\u5927\uff0c\u5e76\u5206\u5316\u5f62\u6210\u4e0d\u540c\u8111\u533a\u7ed3\u6784\uff08\u56fe1A\uff09\u3002HiPSCs\u5b9a\u5411\u5206\u5316\u5f62\u6210\u76843D\u5927\u8111\u7c7b\u5668\u5b98\u80fd\u5728\u4e00\u5b9a\u7a0b\u5ea6\u4e0a\u6a21\u62df\u4eba\u7c7b\u80ce\u513f\u5927\u8111\u53d1\u80b2\u8fdb\u7a0b\u3002\u5728\u5927\u8111\u7c7b\u5668\u5b98\u5206\u5316\u7684\u795e\u7ecf\u5916\u80da\u5c42\u9636\u6bb5\u52a0\u5165\u4e0d\u540c\u6d53\u5ea6\u7684\u841d\u535c\u786b\u7d20\u8fdb\u884c\u5904\u7406\uff0c\u76f8\u6bd4\u4e8e\u5bf9\u7167\u7ec4\uff0c\u5b9e\u9a8c\u7ec4\u663e\u793a\u51fa\u8f83\u5feb\u7684\u751f\u957f\u901f\u5ea6\u3002HE\u67d3\u8272\u793a\u52a0\u5165\u841d\u535c\u786b\u7d2040 d\u5de6\u53f3\u51fa\u73b0\u660e\u663e\u7684\u751f\u957f\uff0c\u57f9\u517b70 d\u540e\u5b9e\u9a8c\u7ec4\u4f53\u79ef\u76f8\u5bf9\u4e8e\u5bf9\u7167\u7ec4\u660e\u663e\u589e\u5927\uff0c\u7ed3\u6784\u66f4\u52a0\u5b8c\u6574\uff1b\u89c1\u56fe1B\u53ca\u56fe2\u3002<\/p>\n<\/p>\n

\u56fe1 1 \u03bcmol\/L\u6d53\u5ea6\u841d\u535c\u786b\u7d20\u5904\u7406\u540e\u4e0d\u540c\u65f6\u95f4\u70b9\u5b9e\u9a8c\u7ec4\uff08n=3\uff09\u548c\u5bf9\u7167\u7ec4\uff08n=3\uff09\u5927\u8111\u7c7b\u5668\u5b98\u7684\u751f\u957f\u60c5\u51b5<\/p>\n<\/p>\n

\u56fe2 \u4e0d\u540c\u6d53\u5ea6\u841d\u535c\u786b\u7d20\u5904\u7406\u540e\u7684\u5b9e\u9a8c\u7ec4\uff08n=3\uff09\u4e0e\u5bf9\u7167\u7ec4\uff08n=3\uff0940 d\u548c70 d\u5927\u8111\u7c7b\u5668\u5b98\u751f\u957f\u60c5\u51b5 HE \u67d3\u8272<\/p>\n

\u4e8c\u3001\u841d\u535c\u786b\u7d20\u4fc3\u8fdb\u5927\u8111\u7c7b\u5668\u5b98\u524d\u989d\u53d1\u80b2\u548c\u795e\u7ecf\u5143\u5206\u5316<\/p>\n

\u514d\u75ab\u8367\u5149\u67d3\u8272\u793a\u57f9\u517b40 d\u540e\uff0c2\u7ec4\u7c7b\u5668\u5b98\u4e2d\u5404\u4e2a\u8111\u533a\u6807\u5fd7\u7269\u5982\u524d\u8111\u6807\u5fd7\u7269FOXG1\u3001\u8111\u5ba4\u6837\u7ed3\u6784\u6807\u5fd7\u7269N-cadherin\u3001\u524d\u989d\u53f6\u76ae\u8d28AUST2\u548c\u6d77\u9a6c\u5e38\u89c1\u6807\u5fd7\u7269Frizzled 9\u5747\u51fa\u73b0\u8868\u8fbe\uff0c\u524d\u8111\u3001\u8111\u5ba4\u6837\u7ed3\u6784\u3001\u524d\u989d\u53f6\u76ae\u8d28\u548c\u6d77\u9a6c\u6837\u7ed3\u6784\u5f62\u6210\u3002\u4e0e\u5bf9\u7167\u7ec4\u5927\u8111\u7c7b\u5668\u5b98\u6bd4\u8f83\uff0c\u52a0\u5165\u841d\u535c\u786b\u7d20\u5904\u7406\uff081 \u03bcmol\/L\uff0940 d\u7684\u5927\u8111\u7c7b\u5668\u5b98\u4e2d\u53ef\u89c1\u524d\u8111\u6807\u5fd7\u86cb\u767dFOXG1\u3001\u795e\u7ecf\u5143\u548c\u76ae\u8d28\u53d1\u80b2\u6807\u5fd7\u86cb\u767dPAX6\u3001\u524d\u989d\u53f6\u76ae\u8d28AUST2\u3001\u795e\u7ecf\u5e72\u7ec6\u80de\u6807\u5fd7\u86cb\u767dSOX2\u8868\u8fbe\u660e\u663e\u589e\u5f3a\uff08\u56fe3\uff09\u3002\u5728\u57f9\u517b70 d\u7684\u5927\u8111\u7c7b\u5668\u5b98\u4e2d\uff0c\u52a0\u5165\u4e0d\u540c\u6d53\u5ea6\u841d\u535c\u786b\u7d20\u5904\u7406\u540e\uff0c\u5927\u8111\u7c7b\u5668\u5b98\u4e2d\u6df1\u5c42\u76ae\u8d28\u6807\u5fd7\u7269TBR1\uff0c\u795e\u7ecf\u5143\u6807\u5fd7\u86cb\u767dTUJ1\u3001NeuN\u3001\u795e\u7ecf\u5fae\u7ba1MAP2\u8868\u8fbe\u589e\u5f3a\uff08\u56fe4\uff09\u3002\u5927\u8111\u7c7b\u5668\u5b98\u7ec6\u80de\u7c7b\u578b\u6807\u5fd7\u7269TUJ1\/GFAP\u7684\u6bd4\u4f8b40 d\u4e3a0.55\uff0c70 d\u4e3a2.57\uff0c70 d\u65f6\u795e\u7ecf\u5143\u7684\u8868\u8fbe\u6bd4\u80f6\u8d28\u7ec6\u80de\u66f4\u591a\u3002<\/p>\n<\/p>\n

\u56fe3 1 \u03bcmol\/L\u6d53\u5ea6\u841d\u535c\u786b\u7d20\u5904\u740640 d\u5b9e\u9a8c\u7ec4\u4e0e\u5bf9\u7167\u7ec4\u5927\u8111\u7c7b\u5668\u5b98SOX2\u3001PAX6\u3001FOXG1\u3001AUST2\u8868\u8fbe\u589e\u5f3a \u514d\u75ab\u8367\u5149\u67d3\u8272<\/p>\n<\/p>\n

\u56fe4 1 \u03bcmol\/L\u6d53\u5ea6\u841d\u535c\u786b\u7d20\u5904\u740670 d\u5b9e\u9a8c\u7ec4\uff08n=3\uff09\u4e0e\u5bf9\u7167\u7ec4\uff08n=3\uff09\u5927\u8111\u7c7b\u5668\u5b98TBR1\u3001TUJ1\u3001NeuN\u3001MAP2\u8868\u8fbe\u589e\u5f3a \u514d\u75ab\u8367\u5149\u67d3\u8272<\/p>\n

\u4e09\u3001\u57fa\u56e0\u6d4b\u5e8f\u751f\u7269\u4fe1\u606f\u5206\u6790<\/p>\n

1.\u5dee\u5f02\u57fa\u56e0\u7b5b\u9009\uff1a\u4f7f\u7528R\u8bed\u8a00edgeR\u5305\u7b5b\u9009\u51fa\u7b26\u5408\u6761\u4ef6\u7684\u5dee\u5f02\u57fa\u56e0\uff08\u4ee5log2\u4e3a\u5e95\u8ba1\u7b97\u6837\u672c\u95f4\u8868\u8fbe\u91cf\u7684\u5dee\u5f02\u500d\u6570\u503c\u2265 2\u6216\u4ee5log2\u4e3a\u5e95\u8ba1\u7b97\u6837\u672c\u95f4\u8868\u8fbe\u91cf\u7684\u5dee\u5f02\u500d\u6570\u503c\u2264-2\uff0cP<0.05\uff09\u3002\u76f8\u6bd4\u4e8e\u5bf9\u7167\u7ec4\uff0c1 \u03bcmol\/L\u841d\u535c\u786b\u7d20\u5904\u7406\u5b9e\u9a8c\u7ec4\u57fa\u56e0\u4e0a\u8c03105\u4e2a\uff0c\u57fa\u56e0\u4e0b\u8c03512\u4e2a\uff0c\u5171617\u4e2a\uff08\u56fe5\uff09\u3002<\/p>\n<\/p>\n

\u56fe5 1 \u03bcmol\/L\u841d\u535c\u786b\u7d20\u5904\u7406\u5b9e\u9a8c\u7ec4\uff08n=3\uff09\u4e0e\u5bf9\u7167\u7ec4\uff08n=3\uff09\u5dee\u5f02\u57fa\u56e0\u7684\u706b\u5c71\u56fe<\/p>\n

2.\u5dee\u5f02\u57fa\u56e0\u7684KEGG\u548cGO\u5206\u6790\uff1aKEGG\u5206\u6790\u663e\u793a\u6392\u540d\u524d10 \u7684\u4e3b\u8981\u5f71\u54cd\u901a\u8def\u89c1\u56fe6\u3002\u5728GO\u901a\u8def\u4e2d\u841d\u535c\u786b\u7d20\u5bf9\u795e\u7ecf\u5143\u5f62\u6001\u6784\u5efa\u3001\u7a81\u89e6\u8fde\u63a5\u8d77\u7740\u91cd\u8981\u4f5c\u7528\uff0c\u8be6\u89c1\u56fe6\u3002<\/p>\n<\/p>\n

\u56fe6 \u5b9e\u9a8c\u7ec4\uff08n=3\uff09\u4e0e\u5bf9\u7167\u7ec4\uff08n=3\uff09\u5927\u8111\u7c7b\u5668\u5b98\u5dee\u5f02\u8868\u8fbe\u57fa\u56e0\u5728KEGG\u53ca\u751f\u7269\u8fc7\u7a0b\u3001\u7ec6\u80de\u7ec4\u5206\u548c\u5206\u5b50\u529f\u80fd\u5c5e\u6027\u65b9\u9762\u7684GO\u5206\u6790<\/p>\n

3.GSEA\u5206\u6790\uff1a\u52a0\u5165\u841d\u535c\u786b\u7d20\u6d4b\u5e8fGSEA\u5bcc\u96c6\u5206\u6790\u65f6\u53d1\u73b0\u7684\u5177\u6709\u4ee3\u8868\u6027\u75be\u75c5\u548c\u751f\u7269\u529f\u80fd\u57fa\u56e0\u96c6\uff1a\uff081\uff09\u7ec6\u80de\u53d1\u80b2\uff08\u5982\u7ec6\u80de\u589e\u6b96\u3001\u795e\u7ecf\u5143\u548c\u7a81\u89e6\u53d1\u80b2\u3001\u8f74\u7d22\u5f62\u6210\u548c\u795e\u7ecf\u7cfb\u7edf\u5206\u5316\uff09\uff1b\uff082\uff09\u795e\u7ecf\u53d1\u80b2\u548c\u529f\u80fd\uff08\u5982\u5927\u8111\u5f62\u6001\u3001\u795e\u7ecf\u5143\u8fc1\u79fb\u3001\u8f74\u7a81\u5206\u652f\u548c\u5f62\u6001\u4ee5\u53ca\u957f\u671f\u7a81\u89e6\u6291\u5236\uff09\uff1b\uff083\uff09\u795e\u7ecf\u75be\u75c5\uff08\u5982\u8ba4\u77e5\u529f\u80fd\u969c\u788d\u3001\u7f3a\u8840\u6027\u4e2d\u98ce\u3001\u766b\u75eb\u53d1\u4f5c\u3001\u795e\u7ecf\u969c\u788d\uff09\uff1b\uff084\uff09\u7ec6\u80de\u95f4\u4fe1\u53f7\u548c\u76f8\u4e92\u4f5c\u7528\uff08\u5982\u795e\u7ecf\u4f20\u9012\u3001\u7535\u4f4d\u548c\u7a81\u89e6\u7ec4\u7ec7\uff09\uff1b\uff085\uff09\u5fc3\u7406\u969c\u788d\uff08\u5982\u7126\u8651\u3001\u667a\u529b\u8fdf\u949d\u3001\u8a00\u8bed\u548c\u8bed\u8a00\u969c\u788d\u3001\u6291\u90c1\uff09\u3002GSEA\u5bcc\u96c6\u5206\u6790\u4e2d\u57fa\u56e0\u8868\u8fbe\u8f83\u5f3a\u7684\u5bcc\u96c6\u56fe\uff1a\u5927\u8111\u76ae\u8d28\u653e\u5c04\u72b6\u7ec6\u80de\u8fc1\u79fb\u57fa\u56e0\u96c6\uff08GO\uff1a0021799\uff09\uff0c\u7ebf\u7c92\u4f53\u751f\u7269\u53d1\u751f\u7684\u8f6c\u5f55\u6fc0\u6d3b\u57fa\u56e0\u96c6\uff08R-HSA-2151201\uff09\u3001NOTCH\u4fe1\u53f7\u901a\u8def\u57fa\u56e0\u96c6\uff08HSA04330\uff09\u3001\u8111\u795e\u7ecf\u75be\u75c5\u57fa\u56e0\u96c6\uff08DOID\uff1a5656\uff09\u3001\u7ec4\u86cb\u767dH4\u4e59\u9170\u5316\u57fa\u56e0\u96c6\uff08GO\uff1a0043967\uff09\uff1b\u89c1\u56fe7\u3002<\/p>\n<\/p>\n

\u56fe7 \u5b9e\u9a8c\u7ec4\uff08n=3\uff09\u4e0e\u5bf9\u7167\u7ec4\uff08n=3\uff09\u5927\u8111\u7c7b\u5668\u5b98\u5dee\u5f02\u8868\u8fbe\u57fa\u56e0GSEA\u5206\u6790<\/p>\n

4. qRT-PCR\u9a8c\u8bc1\u5dee\u5f02\u6027\u8868\u8fbe\u57fa\u56e0\uff1a\u901a\u8fc7qRT-PCR\u5206\u6790\u5dee\u5f02\u8868\u8fbe\u57fa\u56e0\u5728\u5927\u8111\u7c7b\u5668\u5b98\u4e2d\u7684\u8868\u8fbe\u60c5\u51b5\uff0c\u9a8c\u8bc1\u7684\u5dee\u5f02\u8868\u8fbe\u57fa\u56e0\u53ca\u5176\u5f15\u7269\u5e8f\u5217\u89c1\u88681\u3002\u7ed3\u679c\u663e\u793a\uff0c\u4e0e\u5bf9\u7167\u7ec4\u5927\u8111\u7c7b\u5668\u5b98\u76f8\u6bd4\uff0cSFTPC\u3001AKR1C3\u3001CXCR6\u3001PRAP1\u3001TMC8\u3001GPR182\u3001F2RL2\u3001KCNJ10\u57fa\u56e0\u53d8\u5316\u8d8b\u52bf\u4e0e\u6d4b\u5e8f\u7ed3\u679c\u4e00\u81f4\uff0c\u5176\u8f6c\u5f55\u6c34\u5e73\u4e0a\u8c03\uff0c\u5dee\u5f02\u6709\u7edf\u8ba1\u5b66\u610f\u4e49\uff08\u56fe8\uff09\u3002<\/p>\n<\/p>\n

\u56fe8 qRT-PCR\u9a8c\u8bc1\u5b9e\u9a8c\u7ec4\uff08n=3\uff09\u4e0e\u5bf9\u7167\u7ec4\uff08n=3\uff09\u5728\u57fa\u56e0\u8868\u8fbe\u8f6c\u5f55\u6c34\u5e73\u5dee\u5f02\u6709\u7edf\u8ba1\u5b66\u610f\u4e49<\/p>\n

\u8ba8\u8bba<\/p>\n

\u6211\u4eec\u5bf9\u4e0d\u540c\u6d53\u5ea6\u841d\u535c\u786b\u7d20\u5904\u7406\u7684HiPSCs\u6765\u6e90\u7684\u5927\u8111\u7c7b\u5668\u5b98\u6a21\u578b\u8fdb\u884c\u7814\u7a76\uff0c\u63a2\u7d22\u841d\u535c\u786b\u7d20\u5bf9\u5927\u8111\u7c7b\u5668\u5b98\u751f\u957f\u7684\u5f71\u54cd\u53ca\u53ef\u80fd\u7684\u76f8\u5173\u4fe1\u53f7\u901a\u8def\uff0c\u7814\u7a76\u7ed3\u679c\u663e\u793a\u841d\u535c\u786b\u7d20\u5bf9\u5927\u8111\u7c7b\u5668\u5b98\u5728\u7ec4\u7ec7\u3001\u7ec6\u80de\u548c\u57fa\u56e0\u8868\u8fbe\u6c34\u5e73\u4e0a\u6709\u4e00\u5b9a\u7684\u4fc3\u8fdb\u751f\u957f\u548c\u5206\u5316\u4f5c\u7528\u3002\u841d\u535c\u786b\u7d20\u5904\u7406\u5927\u8111\u7c7b\u5668\u5b9840 d\uff0c\u5b9e\u9a8c\u7ec4\u4e0e\u5bf9\u7167\u7ec4\u76f8\u6bd4\uff0c\u5176\u524d\u989d\u53f6\u76ae\u8d28\u3001\u524d\u8111\u3001\u795e\u7ecf\u5e72\u7ec6\u80de\u6807\u5fd7\u86cb\u767d\u8868\u8fbe\u660e\u663e\u589e\u5f3a\uff1b70 d\u65f6\uff0c\u5b9e\u9a8c\u7ec4\u8f83\u5bf9\u7167\u7ec4\u795e\u7ecf\u5143\u8868\u8fbe\u660e\u663e\u589e\u5f3a\u3002\u751f\u7269\u4fe1\u606f\u5b66\u5206\u6790\u8868\u660e\uff0c\u841d\u535c\u786b\u7d20\u5bf9\u5927\u8111\u7c7b\u5668\u5b98\u7684\u4fc3\u8fdb\u4f5c\u7528\u53ef\u80fd\u4e0e\u795e\u7ecf\u5143\u5f62\u6001\u6784\u5efa\u3001\u7a81\u89e6\u8fde\u63a5\u6709\u5173\u3002<\/p>\n

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