Transformation of bone marrow mesenchymal stem cells in three-dimension scaffolds with dynamic environment in vitro

[[abstract]]將人類骨髓間葉幹細胞(hBMSC)植入包括第二型膠原蛋白改質的PLGA/PLLA 支架(BCII)與幾丁聚醣/動物明膠混合支架(CG) 的兩種三維支架，接著以含有TGF-β3 誘導因子之培養基靜態或動態培養23 天後，進行細胞支架中細胞數量、基質含量、細胞型態、細胞分化及機械性質之分析。結果發現在靜態培養下兩種支架中，細胞數量皆會因添加TGF-β3而增加。而不論是否添加TGF-β3，CG支架內的細胞數量較BCII支架為高。以s-100 protein抗體進行組織免疫學分析，在兩種支架中均發現TGF-β3能成功將骨髓間葉幹細胞分化。CG支架在動態培養，並以添加TGF-β3 之培養基進行培養後細胞數量較靜態培養明顯增加37%，但細胞分化數量卻下降。故可歸結出支架種類並不影響TGF-β3 促進骨髓間葉幹細胞之分化；動態培養並不幫助骨髓間葉幹細胞之分化。 Human bone marrow mesenchymal stem cells (hBMSC) were seeded into two scaffolds, including blended polymers of PLGA50/50 and PLLA modified by type II collagen (BCII) and complexes of chitosan and gelatin (CG). Then scaffolds were incubated in culture medium with TGF-β3 for 23 days. Cell number, amount of matrix, cell morphology and cell differentiation in constructs were determined. The cell number in two scaffolds was higher when TGF-β3 was added into culture medium. Cell numbers in CG scaffolds were higher than those in BCII scaffolds. Anti s-100 protein was used as the marker for differentiated cells in constructs. Many differentiated cells in two scaffolds were observed when TGF-β3 was employed. The materials of the scaffolds had no effect on TGF-β3 induced hBMSC transformation into differentiated cells