Efficient differentiation of mouse embryonic stem cells into insulin-producing cells.

Experimental Diabetes Research Pub Date : 2012-01-01 Epub Date: 2012-08-05 DOI:10.1155/2012/201295
Szu-Hsiu Liu, Lain-Tze Lee
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引用次数: 14

Abstract

Embryonic stem (ES) cells are a potential source of a variety of differentiated cells for cell therapy, drug discovery, and toxicology screening. Here, we present an efficacy strategy for the differentiation of mouse ES cells into insulin-producing cells (IPCs) by a two-step differentiation protocol comprising of (i) the formation of definitive endoderm in monolayer culture by activin A, and (ii) this monolayer endoderm being induced to differentiate into IPCs by nicotinamide, insulin, and laminin. Differentiated cells can be obtained within approximately 7 days. The differentiation IPCs combined application of RT-PCR, ELISA, and immunofluorescence to characterize phenotypic and functional properties. In our study, we demonstrated that IPCs produced pancreatic transcription factors, endocrine progenitor marker, definitive endoderm, pancreatic β-cell markers, and Langerhans α and δ cells. The IPCs released insulin in a manner that was dose dependent upon the amount of glucose added. These techniques may be able to be applied to human ES cells, which would have very important ramifications for treating human disease.

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小鼠胚胎干细胞向胰岛素生成细胞的有效分化。
胚胎干细胞是多种分化细胞的潜在来源,可用于细胞治疗、药物发现和毒理学筛选。在这里,我们提出了一种将小鼠胚胎干细胞分化为胰岛素生成细胞(IPCs)的有效策略,通过两步分化方案,包括(i)通过激活素a在单层培养中形成最终内胚层,以及(ii)通过烟酰胺、胰岛素和层粘胶蛋白诱导单层内胚层分化为胰岛素生成细胞。分化细胞可在约7天内获得。分化IPCs联合应用RT-PCR、ELISA和免疫荧光表征表型和功能特性。在我们的研究中,我们证明了IPCs产生胰腺转录因子、内分泌祖细胞标记物、最终内胚层、胰腺β细胞标记物和朗格汉斯α和δ细胞。IPCs以一种剂量依赖于葡萄糖添加量的方式释放胰岛素。这些技术可能应用于人类胚胎干细胞,这将对治疗人类疾病产生非常重要的影响。
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来源期刊
Experimental Diabetes Research
Experimental Diabetes Research 医学-内分泌学与代谢
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审稿时长
3-8 weeks
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