{"id":240,"date":"2025-01-26T22:26:53","date_gmt":"2025-01-26T14:26:53","guid":{"rendered":"https:\/\/tiand.top\/?p=240"},"modified":"2025-01-26T22:38:42","modified_gmt":"2025-01-26T14:38:42","slug":"%e5%b0%8f%e9%bc%a0%e8%83%8c%e6%a0%b9%e7%a5%9e%e7%bb%8f%e8%8a%82%e7%bb%86%e8%83%9e%e7%9a%84%e5%88%86%e7%a6%bb%e5%92%8c%e5%9f%b9%e5%85%bb","status":"publish","type":"post","link":"https:\/\/tiand.top\/index.php\/2025\/01\/26\/%e5%b0%8f%e9%bc%a0%e8%83%8c%e6%a0%b9%e7%a5%9e%e7%bb%8f%e8%8a%82%e7%bb%86%e8%83%9e%e7%9a%84%e5%88%86%e7%a6%bb%e5%92%8c%e5%9f%b9%e5%85%bb\/","title":{"rendered":"\u5c0f\u9f20\u80cc\u6839\u795e\u7ecf\u8282\u7ec6\u80de\u7684\u5206\u79bb\u548c\u57f9\u517b"},"content":{"rendered":"\n

\u3010\u5b9e\u9a8c\u6750\u6599\u3011<\/h4>\n\n\n\n
    \n
  1. 4-6<\/strong>\u5468\u9f84\u5c0f\u9f20\uff08\u4e00\u822c<8\u5468\u9f84\uff09<\/strong><\/li>\n\n\n\n
  2. 40 \u00b5m \u7ec6\u80de\u6ee4\u7f51\uff0c70\u00b5m\u7ec6\u80de\u6ee4\u7f51\uff0c 0.22 \u00b5m\u6ee4\u5668\u3002<\/li>\n\n\n\n
  3. \u5c42\u7c98\u8fde\u86cb\u767d(Sigma, L2020)\uff1a<\/strong>\u5236\u5907 150ul \/\u5b54\uff086\u5b54\u677f\uff0910 \u03bcg\/mL \u6eb6\u6db2\uff081:100 \u7a00\u91ca\uff09\uff0c\u50a8\u5907\u6eb6\u6db2\uff081 mg\/mL \u5c42\u7c98\u8fde\u86cb\u767d\uff0c\u221220\u00b0C \u4e0b\u50a8\u5b58\uff09\u3002\u5728 5 mL PBS (1:100) \u4e2d\u7a00\u91ca 50 \u03bcL 1 mg\/mL \u5c42\u7c98\u8fde\u86cb\u767d\u50a8\u5907\u6eb6\u6db2\uff0c\u5de5\u4f5c\u6d53\u5ea6\u4e3a 10 \u03bcg\/mL\u3002<\/li>\n\n\n\n
  4. \u6df7\u5408\u6d88\u5316\u6db2<\/strong>\uff1a100\u500d\u7a00\u91ca100X collagenase type I\u548c100X dispase II\uff0c \u7ec8\u6d53\u5ea6\uff1a1mg\/ml collagenase type I\u548c5 mg\/ml dispase II\u7684Hanks\u5e73\u8861\u76d0\u6eb6\u6db2 \uff0c\u8fc7\u6ee4\u9664\u83cc<\/strong>\u3002<\/li>\n\n\n\n
  5. \u63d0\u53d6\u57f9\u517b\u57fa\uff1a<\/strong>DMEM\u3002<\/li>\n\n\n\n
  6. DRG<\/strong>\u6df7\u5408\u57f9\u517b\u57fa\uff1a<\/strong>Neurobasal\u57f9\u517b\u57fa(Thermo, 10888022)\u52a02% B-27(Thermo, 17504044)\u300110%\u80ce\u725b\u8840\u6e05\u30011\uff05\u53cc\u6297\u548c1\uff05L-\u8c37\u6c28\u9170\u80fa\uff0c48h\uff0c5ml\/\u5b54\u3002<\/li>\n\n\n\n
  7. DRG<\/strong>\u6362\u6db2\u57f9\u517b\u57fa<\/strong>\uff1a Neurobasal\u57f9\u517b\u57fa+2%B27+1\uff05\u8c37\u6c28\u9170\u80fa\uff0b1\uff05\u53cc\u6297+20ng\/mlNGF\uff08\u539f\u6db21\uff1a5000\uff09+ 50\u00b5M 5-FU+150\u00b5m uridine \uff08\u4ee5\u4e0a\u4e24\u9879\u5728\u57f9\u517b\u65f6\u73b0\u52a0\uff09\u3002<\/li>\n\n\n\n
  8. 100X collagenase type I<\/strong> (Thermo, 17100-017)\uff1a\u50a8\u5b58\u6db2100 mg\/ml\uff0c500mg+5mL Hanks\u7a00\u91ca\uff0c100ul\u52301ml \/tube\u3002-20\u2103\u5b58\u50a8\uff0c\u6709\u6548\u671f6\u4e2a\u6708\u3002<\/li>\n\n\n\n
  9. 100X dispase II<\/strong> (Sigma, D4693)\uff1a\u50a8\u5b58\u6db2500 mg\/ml\uff0c1g+2ml Hanks\u7a00\u91ca\uff0c100ul\u52301ml \/tube. -20\u2103\u5b58\u50a8\uff0c\u6709\u6548\u671f6\u4e2a\u6708\u3002<\/li>\n\n\n\n
  10. 100X 5-FU<\/strong> (Sigma, F0503)\uff1a10mg\u6eb6\u51658ml PBS, \u5f975mM\uff0c\u7a00\u91ca100\u500d\u5f9750\u00b5m\u5de5\u4f5c\u6db2\u3002-20\u2103\u5b58\u50a8\uff0c\u73b0\u7528\u73b0\u52a0<\/strong>\u5165\u57f9\u517b\u57fa\u3002<\/li>\n\n\n\n
  11. 66 X uridine<\/strong> (MCE, HY-B1449):<\/strong> 150\u00b5l\u6eb6\u516510ml\u57f9\u517b\u57fa\uff08\u7a00\u91ca66\u500d\uff09\u5f97150\u00b5m\u5de5\u4f5c\u6db2\u3002-20\u2103\u5b58\u50a8\uff0c\u73b0\u7528\u73b0\u52a0<\/strong>\u5165\u57f9\u517b\u57fa\u3002<\/li>\n\n\n\n
  12. 0.5%<\/strong>\u620a\u5df4\u6bd4\u59a5\u94a0<\/strong>\uff1a0.05g\u6eb6\u4e8e10ml\u751f\u7406\u76d0\u6c34\u3002<\/li>\n<\/ol>\n\n\n\n

    \u3010\u5b9e\u9a8c\u65b9\u6cd5\u3011<\/h4>\n\n\n\n
      \n
    1. \u7ec4\u7ec7\u57f9\u517b\u677f\u7684\u5c42\u7c98\u8fde\u86cb\u767d\u6d82\u5c42\uff1a\u63d0\u524d20min\u5c06 150 \u03bcL 10 \u03bcg\/mL \u5c42\u7c98\u8fde\u86cb\u767d\u6eb6\u6db2\u6ef4\u52306\u5b54\u677f\u6bcf\u4e2a\u5b54\u4e2d\u5fc3\uff0c\u653e\u5165 37\u00b0C \u57f9\u517b\u7bb1\u4e2d\uff0c\u76f4\u81f3\u51c6\u5907\u597d\u5b9e\u9a8c\u3002\u5728\u5b54\u4e4b\u95f4\u6dfb\u52a0\u6c34\u6216 PBS\u3002<\/li>\n\n\n\n
    2. \u5c0f\u9f20\u5907\u76ae\uff1a\u8fc7\u9ebb\u5904\u6b7b\uff0c\u5f85\u5b8c\u5168\u6b7b\u4ea1\u540e\u7f6e\u5165\u63d0\u524d\u9884\u51b7\u768475%\u4e59\u9187\u6eb6\u6db2\u6d78\u6ce1\u6d88\u6bd23~5 min\u3002\uff08\u6ce8\u610f\u526a\u5f00\u4e4b\u540e\u4e0d\u8981\u7528\u4e59\u9187\uff0c\u4f1a\u635f\u4f24\u795e\u7ecf\u7ec6\u80de\uff09<\/li>\n\n\n\n
    3. \u79bb\u65ad\u810a\u67f1\uff1a\u6cbf\u80cc\u90e8\u6b63\u4e2d\u7ebf\u526a\u5f00\u5c0f\u9f20\u76ae\u80a4\uff0c\u5411\u4e24\u4fa7\u949d\u6027\u5206\u79bb\u80cc\u90e8\u7b4b\u819c\uff0c\u624b\u672f\u5200\u7247\u6cbf\u97e7\u5e26\u68d8\u7a81\u4ea4\u754c\u5904\u8fc5\u901f\u79bb\u65ad\u767d\u8272\u97e7\u5e26\uff0c\u968f\u540e\u4f7f\u7528\u5200\u67c4\u5411\u4e24\u4fa7\u949d\u6027\u5206\u79bb\u5c0f\u9f20\u80cc\u90e8\u808c\u8089\u7fa4\uff0c\u9732\u51fa\u808c\u8089\u4e0b\u9644\u7740\u4e8e\u6a2a\u7a81\u7684\u97e7\u5e26\uff0c\u5200\u7247\u5206\u79bb\u540e\uff0c\u8fc5\u901f\u6e05\u7406\u5468\u56f4\u808c\u8089\u7ec4\u7ec7\uff0c\u5200\u7247\u79bb\u65ad\u810a\u67f1\u3002\u810a\u67f1\u9644\u5e26\u90e8\u5206\u808b\u9aa8\u653e\u5165\u542bHBSS\u768410cm\u5927\u76bf\uff0c\u51b0\u4e0a\u653e\u7f6e\u3002<\/li>\n\n\n\n
    4. \u6e05\u7406\u810a\u67f1\uff1a\u4f7f\u7528\u542b\u53cc\u6297PBS\u51b2\u6d17\uff0c\u6e05\u9664\u591a\u4f59\u8840\u6db2\u53ca\u808c\u8089\u3002<\/li>\n\n\n\n
    5. \u5206\u79bbDRG\uff1a\uff08\u4ee5\u4e0b\u64cd\u4f5c\u5747\u5728\u4f53\u89c6\u663e\u5fae\u955c\u4e0b\u64cd\u4f5c\uff09\u6cbf\u6b63\u4e2d\u7ebf\u81ea\u690e\u95f4\u5b54\u5904\u526a\u5f00\u9525\u4f53,\u5206\u4e3a\u5de6\u53f3\u4e24\u534a; \u5fc5\u8981\u65f6\u526a\u53bb\u68d8\u7a81\u4ee5\u65b9\u4fbf\u5206\u79bb\u9525\u4f53\uff0c\u4f7f\u7528\u5438\u5165PBS\u6ce8\u5c04\u5668\u51b2\u6d17\u690e\u95f4\u5b54\uff0c\u6e05\u7406\u591a\u4f59\u8840\u6db2\uff0c\u5373\u53ef\u89c1\u4fa7\u9690\u7a9d\uff1bDRG\u5904\u4e8e\u4fa7\u9690\u7a9d\u5185,\u4e3a\u6de1\u9ec4\u8272\u7ed3\u8282\u72b6\u7ec4\u7ec7\uff0c\u4e24\u7aef\u5747\u6709\u795e\u7ecf\u7ea4\u7ef4\u7ec4\u7ec7,\u4f7f\u7528\u5e26\u9f7f\u773c\u79d1\u663e\u5fae\u954a\u6444\u53d6DRG\u7ec4\u7ec7\uff0c\u8f7b\u8f7b\u63d0\u8d77\u540e\u4f7f\u7528\u663e\u5fae\u89d2\u819c\u526a\uff08\u76f4\u526a\/\u5f2f\u526a\uff09\u526a\u65ad\u4e24\u7aef\u795e\u7ecf\u7ea4\u7ef4\u3002<\/li>\n\n\n\n
    6. \u6536\u96c6DRG\uff1a\u4fee\u7406\u591a\u4f59\u795e\u7ecf\u7ea4\u7ef4\u540e,\u5c06DRG\u7f6e\u4e8e6\u5b54\u677f\u4e2d\u5e76\u8865\u5145 DMEM\uff0c\u7ee7\u7eed\u4ece\u6240\u6709\u690e\u95f4\u5b54\u6536\u96c6\u6bcf\u4e2a DRG\u3002<\/li>\n\n\n\n
    7. \u6d88\u5316\uff1a\u5c06DRG\u572810ml\u6df7\u5408\u6d88\u5316\u9176\u6eb6\u6db2\u4e2d\u5728 37\u00b0C \u57f9\u517b\u7bb1\u4e2d\u6447\u52a8\uff08150 rpm\uff0915 \u5206\u949f\uff0c\u52a01ml 10 % FBS DMEM\uff0c\u5439\u6253\u81f3\u770b\u4e0d\u89c1\u5927\u5757\u7ec4\u7ec7\uff0c\u5c06\u7ec6\u80de\u60ac\u6d6e\u6db2\u901a\u8fc770 \u00b5m \u7ec6\u80de\u6ee4\u7f51\u8fc7\u6ee4\u3002<\/li>\n\n\n\n
    8. \u6d17\u6da4\uff1a\u572825\u00b0C\u4e0b300g\u79bb\u5fc35\u5206\u949f\uff0c\u540e\u5f03\u53bb\u4e0a\u6e05.<\/li>\n\n\n\n
    9. \u91cd\u60ac\uff1a\u5f03\u53bb\u4e0a\u6e05\u6db2\uff0c\u52a0\u51652mL\u6df7\u5408\u57f9\u517b\u57fa\u91cd\u60ac\u6eb6\u6db2\u3002<\/li>\n\n\n\n
    10. \u5236\u5907\u5355\u7ec6\u80de\u60ac\u6db2:\u5300\u901f\u5439\u6253\u5236\u6210\u5355\u7ec6\u80de\u60ac\u6db2\u3002<\/li>\n\n\n\n
    11. \u5c06\u4e0a\u8ff0\u7ec6\u80de\u60ac\u6db2\u752840 \u00b5m \u7ec6\u80de\u6ee4\u7f51\u9664\u53bb\u5927\u5757\u7ea4\u7ef4\uff0c300g\uff0c2min\u79bb\u5fc3\u6765\u83b7\u53d6\u795e\u7ecf\u5143\u3002<\/li>\n\n\n\n
    12. \u5f03\u5c42\u7c98\u8fde\u86cb\u767d\u6eb6\u6db2\uff08\u53ef\u91cd\u590d\u4f7f\u75282\u301c3\u6b21\uff09\uff0c2 mL PBS \u6e05\u6d17\u57f9\u517b\u76bf3\u6b21\uff0c\u75285ml\u6df7\u5408\u57f9\u517b\u57fa\u5c06\u7ec6\u80de\u91cd\u60ac\u540e\u63a5\u79cd\u5230\u5c42\u7c98\u8fde\u86cb\u767d\u5305\u88ab\u8fc7\u76846\u5b54\u677f\u4e2d\uff0c\u653e\u5165\u5b75\u7bb1\u3002<\/li>\n\n\n\n
    13. \u6362\u6db2\uff1a48\u5c0f\u65f6\u540e\uff0c\u66f4\u6362\u4e3a\u8865\u5145\u6362\u6db2\u57f9\u517b\u57fa\uff1b\u6b64\u540e\uff0c\u6bcf3\u5929\u66f4\u6362\u4e00\u6b21\u6362\u6db2\u57f9\u517b\u57fa\u3002<\/li>\n\n\n\n
    14. \u57f9\u517b\u4e09\u5929\u540e\u7ec6\u80de\u72b6\u6001\u8fbe\u5230\u7a33\u5b9a\uff0c\u53ef\u8fdb\u884c\u540e\u7eed\u5b9e\u9a8c\u3002<\/li>\n<\/ol>\n\n\n\n

      \u3010Note\u3011<\/h4>\n\n\n\n
        \n
      1. \u63d0\u53d6\u8fc7\u7a0b\u4e2d\u63d0\u53d6\u57f9\u517b\u57fa\u5168\u7a0b\u7f6e\u4e8e\u51b0\u4e0a\uff0c\u6bcf\u53ea\u5c0f\u9f20\u81f3\u5c11\u63d0\u53d620\u4e2aDRG\u3002<\/li>\n\n\n\n
      2. \u6e05\u7406\u690e\u95f4\u5b54\u5185\u7684\u810a\u9ad3\u7ec4\u7ec7\u4e0e\u795e\u7ecf\u7ea4\u7ef4\u65f6\u6ce8\u610f\u52a8\u4f5c\u8f7b\u67d4\u7f13\u6162\uff0c\u4ee5\u9632\u8bef\u5c06DRG\u8fde\u540c\u810a\u9ad3\u7ec4\u7ec7\u4e0e\u795e\u7ecf\u7ea4\u7ef4\u4e00\u540c\u6e05\u7406\uff1b<\/li>\n\n\n\n
      3. 5-\u6c1f\u5c3f\u5627\u5576\u7528\u4e8e\u6291\u5236\u536b\u661f\u80f6\u8d28\u7ec6\u80de\u589e\u6b96\uff0c\u540c\u65f6\u5bf9\u975e\u589e\u6b96\u6027\u7ec6\u80de\uff08\u795e\u7ecf\u5143\u7ec6\u80de\uff09\u5f71\u54cd\u8f83\u5c0f\u3002<\/li>\n\n\n\n
      4. \u57f9\u517b\u65f6\u5b9c\u752835 mm\u57f9\u517b\u76bf\/6\u5b54\u677f\uff0c\u7ef4\u6301\u795e\u7ecf\u5143\u7ec6\u80de\u4e4b\u95f4\u7684\u7a81\u89e6\u7684\u9ad8\u6548\u8fde\u63a5<\/li>\n\n\n\n
      5. \u5439\u6253\u6b21\u6570\u8d8a\u5c11\uff0c\u5bf9\u4e8e\u7ec6\u80de\u635f\u4f24\u8d8a\u5c0f\u3002<\/li>\n<\/ol>\n\n\n\n
        \"\"<\/figure>\n\n\n\n
        \"\"<\/figure>\n\n\n\n
        \"\"<\/figure>\n\n\n\n
        \n
        \n
        \"\"<\/figure>\n<\/div>\n\n\n\n
        \n
        \"\"<\/figure>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"

        \u3010\u5b9e\u9a8c\u6750\u6599\u3011 \u3010\u5b9e\u9a8c\u65b9\u6cd5\u3011 \u3010Note\u3011<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_jetpack_memberships_contains_paid_content":false,"footnotes":""},"categories":[8],"tags":[],"class_list":["post-240","post","type-post","status-publish","format-standard","hentry","category-protocols"],"jetpack_featured_media_url":"","jetpack_sharing_enabled":true,"_links":{"self":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts\/240","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/comments?post=240"}],"version-history":[{"count":5,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts\/240\/revisions"}],"predecessor-version":[{"id":251,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts\/240\/revisions\/251"}],"wp:attachment":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/media?parent=240"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/categories?post=240"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/tags?post=240"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}