近來研究上細胞移植治療已經廣泛的應用於組織工程及腫瘤治療等領域，而所採用的細胞來源也因不同的治療目的，主要分為自體移植、同種移植、異種移植及細胞株等。但由於細胞一經由針筒方式注射至目標區域後，受限於注射量的限制以及單顆懸浮細胞不易停留於該位置貼附生長，造成治療效果有限。因此本研究目的為以新型的細胞團塊(spheroid)作為體外實驗模型，研究三維空間下細胞團塊的各種生長機制，並藉由溫度敏感型水膠，建立一個細胞不貼的三維多孔培養系統(multi-well culture system)，以控制細胞數形成合適均一的螢光細胞團塊（直徑200μm）。從MTT活性分析驗證，培養一天的細胞團具備最佳的增殖活性;培養時間過長下，反而會因為細胞計劃性死亡（apoptosis），造成活性下降。另外，由live/dead staining及細胞貼附性分析，證實此團塊中間細胞存活率相當高且在通過針頭注射回種於培養皿後，細胞並未受到傷害，而其貼附能力更由細胞本身的EGFP及免疫染色觀察fibronection，都驗證其貼附生長後，細胞形態與平面培養並無差異。另外，我們也以活細胞培養系統，成功的証實單顆懸浮細胞於水膠上不會貼附，而以聚集成團塊細胞的行為生長，此外，亦以此系統發現團塊細胞依舊保有回貼生長的能力。最後我們以免疫螢光染色，發現細胞間的連結蛋白E-cadherin，培養一天情況下，廣泛的分佈於細胞間，推測其乃是幫助鬆散的細胞團塊逐漸變為緊密的主要因素。另外，也發現團塊表面帶有豐富的ECM蛋白，fibronection、laminin、collagen type I、collagen type III等，有助於體內注射後，在目標位置滯留而貼附生長，改善細胞治療中細胞容易流失，不易停留於目標位置的缺點，有效提昇細胞治療的效果。

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