Limited capacity of human adult islets expanded in vitro to redifferentiate into insulin-producing β-cells
Limited capacity of human adult islets expanded in vitro to redifferentiate into insulin-producing β-cells
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DOI：
10.2337/db06-1545
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发表时间：
2007-03-01
期刊：
影响因子：
7.7
通讯作者：
Hayek, Alberto
中科院分区：
文献类型：
--
作者：
Kayali, Ayse G.;Flores, Luis E.;Hayek, Alberto
有限的器官可用性是胰岛移植在1型糖尿病患者中广泛使用的障碍。为了解决这个问题，许多研究已经探索了体外扩增功能性人类胰岛用于糖尿病细胞治疗的方法。我们以前表明，胰岛细胞复制后，单层形成的影响下，肝细胞生长因子和选定的细胞外基质。然而，在这些条件下，衰老和胰岛素表达的丧失发生在> 15天后。相比之下，其他研究小组报告称，在单层中扩增数月的胰岛细胞通过可逆的上皮-间充质转化进展，并且在从培养物中去除血清后，形成了产生胰岛素的胰岛样结构（1）。本研究的目的是比较两种方法的胰岛扩张免疫染色，实时定量PCR和微阵列在以下时间点：抵达后，单层扩张后，1周后，在无血清培养基。此时，将细胞等分试样移植到裸鼠中以研究体内功能。两种方法在胰岛细胞扩增中显示出相似的结果。通过两种方法扩增后细胞分化的尝试未能一致地恢复β细胞表型。扩增后β细胞的再分化仍然是一个需要解决方案的挑战。
Limited organ availability is an obstacle to the widespread use of islet transplantation in type 1 diabetic patients. To address this problem, many studies have explored methods for expanding functional human islets in vitro for diabetes cell therapy. We previously showed that islet cells replicate after monolayer formation under the influence of hepatocyte growth factor and selected extracellular matrices. However, under these conditions, senescence and loss of insulin expression occur after > 15 doublings. In contrast, other groups have reported that islet cells expanded in monolayers for months progressed through a reversible epithelial-to-mesenchymal transition, and that on removal of serum from the cultures, islet-like structures producing insulin were formed (1). The aim of the current study was to compare the two methods for islet expansion using immunostaining, real-time quantitative PCR, and microarrays at the following time points: on arrival, after monolayer expansion, and after 1 week in serum-free media. At this time, cell aliquots were grafted into nude mice to study in vivo function. The two methods showed similar results in islet cell expansion. Attempts at cell differentiation after expansion by both methods failed to consistently recover a beta-cell phenotype. Redifferentiation of beta-cells after expansion is still a challenge in need of a solution.