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class="art-title">ABW基質膠在類器官(PDO)培養實驗中的應用</h1> <div class="art-info"> 發布時間： 2022-06-07　　點擊次數： 451次 </div> <div class="art-body"> 　　<a href="http://www.51player.com/ParentList-2335935.html" target="_blank">ABW基質膠</a>是從富含胞外基質蛋白的小鼠腫瘤細胞中提取出基底膜基質，其主要成分有層粘連蛋白、Ⅳ型膠原、巢蛋白、硫酸肝素糖蛋白，還包含生長因子和基質金屬蛋白酶等。在室溫條件下，Matrigengel聚合形成具有生物學活性的三維基質，模擬體內細胞基底膜的結構、組成、物理特性和功能，有利于體外細胞的培養和分化，可用于對細胞形態、生化功能、遷移、侵染和基因表達等的研究。<div>　　</div><div>　　類器官是一個在Matrigengel的支撐下形成三維“微器官模型”，可以模擬天然器官的結構和功能。與傳統的二維培養模型相比，類器官可以模擬出整個生物體的生理過程，具有更接近體內細胞組成和行為，基因組更穩定，更適合生物轉染和高通量篩選的優點。與動物模型相比，類器官模型的操作不僅更簡單，而且快速成型，兩到三周即可得到所需數據。</div><div>　　一、實驗材料：</div><div>　　細胞：病人腫瘤細胞</div><div>　　培養基：F12K/DMEM培養基，PDO培養基</div><div>　　耗材：預冷槍頭、預熱24孔板、預冷0.5ml離心管</div><div>　　其它：Matrigengel（ABW貨號0827035）</div><div>　　</div><div>　　二、實驗步驟：</div><div>　　A.實驗準備</div><div>　　1.提前24h將Matrigengel放置于4°冰箱慢慢融化，(為避免反復凍融，收到貨以后分裝成小規格保存，于-20°保存)</div><div>　　2.需要提前制冰。</div><div>　　B.細胞獲?。簭尼t院將病人的腫瘤組織，快速按照獲取原代細胞的方法操作，獲得腫瘤細胞。</div><div>　　C.細胞種板：</div><div>　　1.按每孔5000個細胞稀釋，將細胞懸液放置在冰上，按照比例加入Matrigengel，混合均勻，將混合液滴入10ul-20ul到24孔板底部，每孔約3-4滴。</div><div>　　2.將培養板放入37°培養箱中，孵育30min，待Matrigengel凝固。</div><div>　　3.準備所需量PDO培養基，放入37°水浴鍋預熱。</div><div>　　4.膠滴凝固以后，每孔加入500ul培養基。</div><div>　　5.將培養板放會培養箱中繼續培養。</div><div>　　6.每隔兩天采集圖像記錄。</div> 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