Investigating the suspension culture on aggregation and function of mouse pancreatic β-cells.
Investigating the suspension culture on aggregation and function of mouse pancreatic β-cells.
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研究悬浮培养物对小鼠胰腺β细胞聚集和功能的影响。
DOI：
10.1002/jbm.a.34547
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Hsuan
中科院分区：
文献类型：
--
作者：
Kai;Chang‐Chin Wu;Shu;C. Chiu;S. Sumi;Hsuan
胰岛的完整性和层次结构极大地影响β细胞的生理学。能够维持或改善β细胞聚集的培养基质将有利于糖尿病患者的细胞治疗。在这项研究中，使用未经处理的 IV 型胶原蛋白、Lipidure 和超低附着培养皿来培养鼠 β 细胞系 MIN-6。研究了伪胰岛的形成和生物学性能。结果表明，β 细胞在未经处理的培养皿上形成松散且不规则的聚集体。相反，假胰岛在其他三种基质上形成。 Lipidure和IV型胶原培养皿上的大多数假胰岛直径在100-150μm之间，存活率较高，而在超低附着培养皿上发现了中央严重坏死的大假胰岛（>250μm）。蛋白质印迹分析显示，相对于单细胞，假胰岛具有相对较高的连接蛋白 36 蛋白产量。葡萄糖刺激胰岛素分泌试验显示，IV型胶原上的假胰岛具有较高的刺激指数。来自 TCPS 培养皿的单层细胞和来自 IV 型胶原或 Lipidure 培养皿的假胰岛被进一步移植到糖尿病小鼠中。接受单细胞和伪胰岛的动物血糖水平降低并恢复了体重。组织学检查显示所有植入物均成功植入，胰岛素染色呈阳性。有趣的是，腹膜内葡萄糖耐量试验的曲线下面积显示假胰岛具有较高的葡萄糖消失率。这项研究表明，可以在 IV 型胶原或 Lipidure 培养皿上培养分离的胰岛或产生胰岛素的细胞，以在移植前改善/保持完整性。
The integrity and hierarchical structure of islet influence β-cells physiology dramatically. A culture substrate which can maintain or improve β-cells aggregation shall benefit cell therapy for diabetics. In this study, nontreated, type IV collagen, Lipidure, and ultralow attachment dishes were used to culture a murine β-cell line, MIN-6. The formation and biological performances of pseudoislets were investigated. Results showed that β-cells formed loose and irregular aggregates on nontreated dishes. Oppositely, pseudoislets formed on other three substrates. Most pseudoislets on Lipidure and type IV collagen dishes had a diameter between 100-150 μm with high survival rate, while large pseudoislets (>250 μm) with seriously central necrosis were found on ultralow attachment dishes. Western blot analysis revealed that pseudoislets had relatively higher connexin 36 protein productions relative to single cells. The glucose-stimulated insulin secretion test showed pseudoislets on type IV collagen have high stimulation index. Monolayers from TCPS dishes and pseudoislets from type IV collagen or Lipidure dishes were further transplanted into diabetic mice. Animals received both single cells and pseudoislets had decreasing blood glucose level and regained body weight. Histologic examination revealed that all implants successfully engrafted with positive insulin staining. Interestingly, the area under curve for the intraperitoneal glucose tolerance test showed pseudoislets had superior glucose disappearance rate. This study reveals that isolated islets or insulin-producing cells can be cultured on type IV collagen or Lipidure dishes to improve/maintain integrity prior to transplantation.