Wake Forest Institute for Regenerative Medicine, Wake Forest Baptist Medical Center, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
Wake Forest Institute for Regenerative Medicine, Wake Forest Baptist Medical Center, Medical Center Boulevard, Winston-Salem, NC 27157, USA; Virginia Tech, Wake Forest University School of Biomedical Engineering and Sciences, 320 ICTAS, Stanger St., Virginia Tech, Blacksburg, VA 24060, USA.
J Diabetes Res. 2016;2016:5807876. doi: 10.1155/2016/5807876. Epub 2016 Dec 6.
-Cell replacement through transplantation is the only curative treatment to establish a long-term stable euglycemia in diabetic patients. Owing to the shortage of donor tissue, attempts are being made to develop alternative sources of insulin-secreting cells. Stem cells differentiation and reprograming as well as isolating pancreatic progenitors from different sources are some examples; however, no approach has yet yielded a clinically relevant solution. Dissociated islet cells that are cultured in cell numbers by proliferation provide a promising platform for redifferentiation towards -cells phenotype. In this study, we cultured islet-derived cells and examined the expression of -cell genes during the proliferation. Islets were isolated from porcine pancreases and enzymatically digested to dissociate the component cells. The cells proliferated well in tissue culture plates and were subcultured for no more than 5 passages. Only 10% of insulin expression, as measured by PCR, was preserved in each passage. High glucose media enhanced insulin expression by about 4-18 fold, suggesting a glucose-dependent effect in the proliferated islet-derived cells. The islet-derived cells also expressed other pancreatic genes such as Pdx1, NeuroD, glucagon, and somatostatin. Taken together, these results indicate that pancreatic islet-derived cells, proliferated , retained the expression capacity for key pancreatic genes, thus suggesting that the cells may be redifferentiated into insulin-secreting -like cells.
通过移植细胞进行细胞替代是唯一能够使糖尿病患者长期稳定血糖的治愈性治疗方法。由于供体组织短缺，人们正在尝试开发替代的胰岛素分泌细胞来源。干细胞分化和重编程以及从不同来源分离胰腺祖细胞就是其中的一些例子；然而，迄今为止，还没有一种方法产生了具有临床相关性的解决方案。通过增殖培养的分离胰岛细胞为胰岛细胞表型的再分化提供了一个很有前途的平台。在这项研究中，我们培养了胰岛衍生细胞，并在增殖过程中检测了β细胞基因的表达。我们从猪胰腺中分离胰岛并进行酶消化以分离成分细胞。这些细胞在组织培养板中增殖良好，传代不超过 5 代。每个传代中只有 10%的胰岛素表达（通过 PCR 测量）得以保留。高葡萄糖培养基使胰岛素的表达增强了约 4-18 倍，这表明增殖的胰岛衍生细胞中存在葡萄糖依赖性效应。胰岛衍生细胞还表达了其他胰腺基因，如 Pdx1、NeuroD、胰高血糖素和生长抑素。综上所述，这些结果表明，增殖的胰岛衍生细胞保留了关键胰腺基因的表达能力，因此提示这些细胞可能被重分化为胰岛素分泌样细胞。
