Cell-Matrix Interactions Improve β-Cell Survival and Insulin Secretion in Three-Dimensional Culture
Cell-Matrix Interactions Improve β-Cell Survival and Insulin Secretion in Three-Dimensional Culture
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DOI：
10.1089/ten.tea.2007.0238
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发表时间：
2008-12-01
影响因子：
4.1
通讯作者：
Anseth, Kristi S.
中科院分区：
文献类型：
--
作者：
Weber, Laney M.;Hayda, Kirsten N.;Anseth, Kristi S.
受控的基质相互作用在聚（乙二醇）水凝胶中的三维培养物中呈现给胰腺β细胞。将分散的MIN6 B细胞封装在包含以下细胞外基质（ECM）蛋白的凝胶环境中：I型胶原蛋白，IV型胶原蛋白，IV型，纤维蛋白原，纤维蛋白，层粘连蛋白，层蛋白和玻璃纤维。在含ECM的凝胶中，β细胞存活率明显优于10天内没有ECM的凝胶。相应地，在每个基质蛋白的存在下，封装的β细胞中的凋亡较少，这表明单个基质相互作用可以防止基质信号传导相关的细胞凋亡（Anoikis）。在所有其他实验条件下，在含有IV胶原型胶原蛋白或层粘连蛋白的凝胶中培养的MIN6β细胞对葡萄糖刺激的响应比β细胞分泌更多的胰岛素。胶原型IV型或10 mu g/ml和250 mu g/ml之间的胶原蛋白浓度的变化不会影响胰岛素分泌。最后，呈现IV型胶原蛋白和层粘连蛋白的水凝胶中的β细胞功能揭示了协同相互作用。总蛋白质浓度为100 mU g/ml，测试了三种胶原蛋白与层粘连蛋白不同比率的凝胶组成（25:75、50:50和75:25）。在存在25 mu g/mL的IV型胶原蛋白和75 mu g/ml的层粘连蛋白的情况下，β细胞胰岛素分泌比单独的层粘连蛋白或胶原蛋白大。这些结果表明，特定的，合理设计的细胞外环境促进了孤立的β细胞存活和功能。
Controlled matrix interactions were presented to pancreatic beta-cells in three-dimensional culture within poly (ethylene glycol) hydrogels. Dispersed MIN6 b-cells were encapsulated in gel environments containing the following entrapped extracellular matrix (ECM) proteins:collagen type I, collagen type IV, fibrinogen, fibronectin, laminin, and vitronectin. In ECM-containing gels, beta-cell survival was significantly better than in gels without ECM over 10 days. Correspondingly, apoptosis in encapsulated beta-cells was less in the presence of each matrix protein, suggesting the ability of individual matrix interactions to prevent matrix signaling-related apoptosis (anoikis). MIN6 beta-cells cultured in gels containing collagen type IV or laminin secreted more insulin in response to glucose stimulation than beta-cells in all other experimental conditions. Variations in collagen type IV or laminin concentration between 10 mu g/mL and 250 mu g/mL did not affect insulin secretion. Finally, beta-cell function in hydrogels presenting both collagen type IV and laminin revealed synergistic interactions. With a total protein concentration of 100 mu g/mL, three gel compositions of varying ratios of collagen type IV to laminin (25:75, 50:50, and 75:25) were tested. In the presence of 25 mu g/mL of collagen type IV and 75 mu g/mL of laminin, beta-cell insulin secretion was greater than with laminin or collagen type IV individually. These results demonstrate that specific, rationally designed extracellular environments promote isolated beta-cell survival and function.