A new method for generating insulin-secreting cells from human pancreatic epithelial cells after islet isolation transformed by NeuroD1.
A new method for generating insulin-secreting cells from human pancreatic epithelial cells after islet isolation transformed by NeuroD1.
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DOI：
10.1089/hgtb.2013.122
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发表时间：
2014-05
影响因子：
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通讯作者：
M. Shimoda;Shuyuan Chen;H. Noguchi;M. Takita;K. Sugimoto;T. Itoh;D. Chujo;S. Iwahashi;B. Naziruddin;M. Levy;S. Matsumoto;P. Grayburn
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文献类型：
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作者：
M. Shimoda;Shuyuan Chen;H. Noguchi;M. Takita;K. Sugimoto;T. Itoh;D. Chujo;S. Iwahashi;B. Naziruddin;M. Levy;S. Matsumoto;P. Grayburn
从非内分泌胰腺上皮细胞（NEPEC）产生的胰岛素分泌细胞已被证明在治疗糖尿病中具有潜在的临床用途。然而，以前的方法要么疗效有限，要么需要病毒载体，这阻碍了临床应用。本研究的目的是建立一种无病毒载体的NEPEC胰岛素分泌细胞培养方法。我们使用来自脑死亡供体的研究级人类胰腺和全胰腺切除术后的临床胰腺的非胰岛部分自体胰岛移植治疗慢性胰腺炎。值得注意的是，一些胰岛可以混合在非胰岛部分中，但它们的影响可能有限。NeuroD1基因被诱导到NEPEC使用一个有效的三重脂质体转染方法没有病毒载体，产生胰岛素分泌细胞。通过向培养基中加入生长因子混合物来促进分化。使用研究级人类胰腺，这种有效的方法在NEPEC分化为胰岛素阳性细胞方面表现出很高的效率，这些细胞在葡萄糖刺激下分泌胰岛素，并在移植到糖尿病无胸腺小鼠后改善糖尿病。即使是慢性胰腺炎的胰腺，也能获得相似的疗效。总之，我们采用三重脂质转染方法的有效分化方案使我们能够在没有病毒载体的情况下从人NEPEC获得非常有效的胰岛素分泌细胞。该方法为同种异体和自体胰岛移植提供了补充胰岛素分泌细胞移植的可能性。
The generation of insulin-secreting cells from nonendocrine pancreatic epithelial cells (NEPEC) has been demonstrated for potential clinical use in the treatment of diabetes. However, previous methods either had limited efficacy or required viral vectors, which hinder clinical application. In this study, we aimed to establish an efficient method of insulin-secreting cell generation from NEPEC without viral vectors. We used nonislet fractions from both research-grade human pancreata from brain-dead donors and clinical pancreata after total pancreatectomy with autologous islet transplantation to treat chronic pancreatitis. It is of note that a few islets could be mingled in the nonislet fractions, but their influence could be limited. The NeuroD1 gene was induced into NEPEC using an effective triple lipofection method without viral vectors to generate insulin-secreting cells. The differentiation was promoted by adding a growth factor cocktail into the culture medium. Using the research-grade human pancreata, the effective method showed high efficacy in the differentiation of NEPEC into insulin-positive cells that secreted insulin in response to a glucose challenge and improved diabetes after being transplanted into diabetic athymic mice. Using the clinical pancreata, similar efficacy was obtained, even though those pancreata suffered chronic pancreatitis. In conclusion, our effective differentiation protocol with triple lipofection method enabled us to achieve very efficient insulin-secreting cell generation from human NEPEC without viral vectors. This method offers the potential for supplemental insulin-secreting cell transplantation for both allogeneic and autologous islet transplantation.