Redox-mediated enrichment of self-renewing adult human pancreatic cells that possess endocrine differentiation potential
Redox-mediated enrichment of self-renewing adult human pancreatic cells that possess endocrine differentiation potential
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DOI：
10.1097/00006676-200410000-00015
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发表时间：
2004-10-01
期刊：
影响因子：
2.9
通讯作者：
Olson, LK
中科院分区：
文献类型：
--
作者：
Linning, KD;Tai, MH;Olson, LK
目的：可移植的人类胰岛的有限可获得性刺激了分离能够自我更新和内分泌分化的成人胰腺干/祖细胞的方法的发展。本研究的目的是确定N-乙酰-L半胱氨酸(NAC)对细胞内氧化还原状态的调节是否允许成人胰腺组织中的胰腺干/祖细胞增殖。方法：采用添加NAC的无血清低钙培养液从人胰腺组织中扩增细胞，并检测其在不同生长条件下的分化能力。结果：人胰腺细胞培养共表达α-淀粉酶、白蛋白、波形蛋白和巢蛋白。然而，HPC培养没有表达与分化的胰腺外分泌、导管或内分泌细胞相关的其他基因。许多参与内分泌细胞发育的转录因子，包括Beta 2、Islet-1、Nkx6.1、Pax4和Pax6在HPC培养中都有不同程度的表达。相比之下，胰腺十二指肠同源异型盒因子1(PDX-1)的表达极低，有时检测不到。在HPC中过表达PDX-1可刺激生长抑素、胰升糖素和碳酸氢酶的表达，但对胰岛素基因的表达无影响。HPC可形成三维的胰岛样细胞聚集体，这与生长抑素和胰升糖素的表达有关，但与胰岛素无关。在添加烟酰胺、Exendin-4和/或磷脂酰肌醇-3激酶抑制剂LY294002的分化培养液中培养的HPC，可刺激胰岛素mRNA和蛋白的表达。结论：细胞内氧化还原调节用于丰富具有自我更新和内分泌分化能力的胰腺干/祖细胞。
Objectives: The limited availability of transplantable human islets has stimulated the development of methods needed to isolate adult pancreatic stem/progenitor cells capable of self-renewal and endocrine differentiation. The objective of this study was to determine whether modulation of intracellular redox state with N-acetyl-L-cysteine (NAC) would allow for the propagation of pancreatic stem/progenitor cells from adult human pancreatic tissue.Methods: Cells were propagated from human pancreatic tissue using a serum-free, low-calcium medium supplemented with NAC and tested for their ability to differentiate when cultured under different growth conditions.Results: Human pancreatic cell (HPC) cultures coexpressed alpha-amylase, albumin, vimentin, and nestin. The HPC cultures, however, did not express other genes associated with differentiated pancreatic exocrine, duct, or endocrine cells. A number of transcription factors involved in endocrine cell development including Beta 2, Islet-1, Nkx6.1, Pax4, and Pax6 were expressed at variable levels in HPC cultures. In contrast, pancreatic duodenal homeobox factor 1 (Pdx-1) expression was extremely low and at times undetectable. Overexpression of Pdx-1 in HPC cultures stimulated somatostatin, glucagon, and carbonic anhydrase expression but had no effect on insulin gene expression. HPC cultures could form 3-dimensional islet-like cell aggregates, and this was associated with expression of somatostatin and glucagon but not insulin. Cultivation of HPCs in a differentiation medium supplemented with nicotinamide, exendin-4, and/or LY294002, an inhibitor of phosphatidylinositol-3 kinase, stimulated expression of insulin mRNA and protein.Conclusion: These data support the use of intracellular redox modulation for the enrichment of pancreatic stem/progenitor cells capable of self-renewal and endocrine differentiation.