Edamura Kazuya, Nasu Koko, Iwami Yukiko, Ogawa Hiroyuki, Sasaki Nobuo, Ohgawara Hisako
Division of Cell Replacement and Regenerative Medicine, Medical Research Institute, School of Medicine, Tokyo Women's Medical University, Japan.
Cell Transplant. 2003;12(4):439-46. doi: 10.3727/000000003108746867.
The effect of either adhesion or collagen molecules on cell attachment, insulin secretion, and glucose responsiveness was investigated in adult porcine pancreatic endocrine (PE) cells that were cultured for a longer term in vitro. Six different types of molecules--laminin, fibronectin, poly-L-lysine (PLL), type I collagen, gelatin, and Matrigel--were used. Approximately 2.0 x 10(5) cells per dish of each molecule type were cultured for 4 weeks. In the laminin group, the insulin accumulation was maintained at a significantly higher level than in the control group at 4 weeks of culture, and glucose-stimulated insulin secretion and the insulin-positive rate were also higher than in the control group. In the Matrigel group, islet-like cell clusters were formed, but insulin accumulation rapidly decreased at 3-4 weeks of culture. A large number of PE cells attached tightly and spread in the fibronectin group until the fourth week of culture, but their function was not better than those in the control group. In the PLL and gelatin groups, the PE cell function was not significantly different from that of the control group. In the type I collagen group, insulin secretion was inferior to that of the other groups. The results of this study suggest that laminin is the most suitable extracellular matrix for the long-term culture preservation of PE cells.
在体外长期培养的成年猪胰腺内分泌(PE)细胞中,研究了黏附分子或胶原蛋白分子对细胞附着、胰岛素分泌和葡萄糖反应性的影响。使用了六种不同类型的分子——层粘连蛋白、纤连蛋白、聚-L-赖氨酸(PLL)、I型胶原蛋白、明胶和基质胶。每种分子类型的每皿约2.0×10⁵个细胞培养4周。在层粘连蛋白组中,培养4周时胰岛素积累水平显著高于对照组,葡萄糖刺激的胰岛素分泌和胰岛素阳性率也高于对照组。在基质胶组中,形成了胰岛样细胞簇,但培养3 - 4周时胰岛素积累迅速下降。在纤连蛋白组中,大量PE细胞紧密附着并铺展至培养第四周,但其功能并不优于对照组。在PLL和明胶组中,PE细胞功能与对照组无显著差异。在I型胶原蛋白组中,胰岛素分泌低于其他组。本研究结果表明,层粘连蛋白是PE细胞长期培养保存最合适的细胞外基质。