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西亚试剂:培养胰腺细胞的新方法

胰岛B细胞,即胰岛β细胞,是胰岛细胞的一种,属内分泌细胞的一种,能分泌胰岛素,与胰岛A细胞分泌的胰高血糖素一起起到调节血糖的作用。胰岛B细胞功能受损、胰岛素分泌绝对或相对不足(胰岛素抵抗),会使血糖升高,从而引发糖尿病。

 

相比于以往实验室培养的单细胞而言,三维培养的胰腺β细胞能存活更长的时间、分泌更多的胰岛素。这项新技术有利于研究胰腺疾病如糖尿病和测试新疗法。研究结果发表在TissueEngineering,PartC,Methods杂志上。

实验室培养胰腺细胞是一项具有挑战性的工作,部分原因是要想胰腺细胞存活下来以及正常工作必须要胰腺细胞之间能相互接触。美国科罗拉多大学博尔德分校、印第安纳大学-印第安纳波利斯的AbigailBernard、Chien-ChiLin博士和KristiAnseth博士开发出一种新方法,即使用光刻法创建微孔细胞培养的环境,该环境有利于形成三维胰岛β-细胞团,并且能控制细胞聚集体的大小。研究人员描述这项新技术能从微孔中去除这些细胞团,能将细胞团封装入凝胶中以便测试和种植。(生物谷:Bioon)

 

doi:10.1089/ten.tec.2011.0504
PMC:
PMID:
A Microwell Cell Culture Platform for the Aggregation of Pancreatic β-Cells.


Abigail B. Bernard, Chien-Chi Lin, Kristi S. Anseth.

Cell–cell contact between pancreatic β-cells is important for maintaining survival and normal insulin secretion. Various techniques have been developed to promote cell–cell contact between β-cells, but a simple yet robust method that affords precise control over three-dimensional (3D) β-cell cluster size has not been demonstrated. To address this need, we developed a poly(ethylene glycol) (PEG) hydrogel microwell platform using photolithography. This microwell cell-culture platform promotes the formation of 3D β-cell aggregates of defined sizes from 25 to 210?μm in diameter. Using this platform, mouse insulinoma 6 (MIN6) β-cells formed aggregates with cell–cell adherin junctions. These naturally formed cell aggregates with controllable sizes can be removed from the microwells for macroencapsulation, implantation, or other biological assays. When removed and subsequently encapsulated in PEG hydrogels, the aggregated cell clusters demonstrated improved cellular viability (>90%) over 7 days in culture, while the β-cells encapsulated as single cells maintained only 20% viability. Aggregated MIN6 cells also exhibited more than fourfold higher insulin secretion in response to a glucose challenge compared with encapsulated single β-cells. Further, the cell aggregates stained positively for E-cadherin, indicative of the formation of cell junctions. Using this hydrogel microwell cell-culture method, viable and functional β-cell aggregates of specific sizes were created, providing a platform from which other biologically relevant questions may be answered.