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class="art-info"> 發布時間： 2022-10-19　　點擊次數： 447次 </div> <div class="art-body"> 　　<a href="http://www.51player.com/Products-36775386.html" target="_blank">082703基質膠</a>是從小鼠腫瘤組織提取的基底膜成分（基底膜是動物體內上皮細胞基底面的一層基質膜），所形成的基質膠。主要成分為層粘連蛋白，Ⅳ型膠原蛋白，巢蛋白。同時，基質膠也包含多種生長因子，例如表皮生長因子EFG，血小板衍生生長因子PDGF，神經生長因子NGF等等。<div>　　</div><div>　　操作方法：</div><div>　　一、類器官培養（操作所需時間為1小時）</div><div>　　1、將基質膠于4°C冰箱里的冰上過夜解凍。</div><div>　　2、將24孔板放置于培養箱預熱。</div><div>　　3、用預冷的槍頭將解凍的基質膠進行分裝。</div><div>　　4、準備病人或者動物組織（或者類器官）來源的，細胞總數為1X105的單細胞懸液。300g離心5min。</div><div>　　5、取50µL的OrganoGel基質膠與離心所得的細胞進行混勻。</div><div>　　6、取出預熱的24孔板，立即將基質膠與細胞的混合物滴加在孔板里，立即將24孔板放入培養箱。</div><div>　　7、約10min后，基質膠凝固后，向孔板里加入500µL類器官培養基，放于培養箱進行培養。</div><div>　　</div><div>　　二、腫瘤類器官藥敏實驗（操作所需時間為2小時）</div><div>　　1、將擴增好足夠量的腫瘤類器官（實驗組）以及正常類器官（對照組）用4°C預冷的基礎培養基進行重懸，緩慢機械吹打，促進膠液化溶解，并保持類器官結構完整（可用于懸浮培養于有5%基質膠溶液的基質膠表面）（Guillenetal.2022）?；蛘咄ㄟ^TryplE酶消化獲得單細胞懸液。</div><div>　　2、離心收集類器官細胞，并進行細胞計數。</div><div>　　3、加入OrganoGel基質膠原液，與細胞進行混勻。</div><div>　　4、將細胞與膠的混合物，通過排槍加入37°C預熱過的96孔板，或者384孔板，立即將孔板放入培養箱。</div><div>　　5、大約10min后，OrganoGel基質膠將凝固，加入相應體積的類器官培養液進行培養。</div><div>　　6、類器官形成后（如果是機械吹打，類器官將在傳代24h后就會形成；如果是酶消化，類器官會在3-5天后形成），每個孔分別加入含有PI染料以及不同種類、不同濃度的待篩選的抗腫瘤藥物。</div><div>　　7、用高內涵顯微鏡上進行活細胞成像，測定腫瘤類器官對各種藥物的敏感性。</div> 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