National Center for cell Science, Ganeshkhind, Pune, India.
Islets. 2010 Mar-Apr;2(2):112-20. doi: 10.4161/isl.2.2.11280.
The scarcity of islets for transplantation calls for an alternative sources of islets. The human umbilical cord has been shown to be a reservoir of multipotent stem cells with capacity to differentiate into ectodermal, mesodermal and endodermal lineages. The present investigation deals with isolation and characterization of mesenchymal stem sells (MSC) derived from human umbilical cord and their differentiation into functional islets. Since these MSCs were found to constitutively express nestin we hypothesized that these would be ideal candidates for islet neogenesis without any further manipulation. Human umbilical cord matrix stem cells (hUCMSCs) were found to express CD29, CD44, CD73, CD90, CD105, smooth muscle actin, nestin, vimentin, proliferation marker Ki67 and embryonic markers Oct4, SSEA4. These were found to be negative for CD33, CD34, CD45 and HLA DR. Human UCMSCs exhibited high proliferating capacity for extended period indicating potential for scaling up. When subjected to a cocktail of specific differentiating factors, these cells differentiated into fat, cartilage, bone, neurons and islet like clusters (ILCs). These ILCs stained positive for diphenylthiocarbazone (DTZ) and expressed human C-peptide, insulin and glucagon. Real time qPCR analysis of newly generated islets further demonstrated abundance of Pdx-1, Ngn3, insulin, glucagon and somatostatin transcripts. On transplantation in experimental diabetic mice these ILCs restored normoglycemia, body weight and exhibited normal glucose tolerance test indicating their functional status. Thus, the present study demonstrates potential of constitutively expressing nestin positive progenitor from umbilical cord as a novel source for islet neogenesis and their usage in cell replacement therapy for diabetes.
胰岛供体的匮乏促使人们寻求胰岛的替代来源。人脐带被证实是多能干细胞的储存库，具有分化为外胚层、中胚层和内胚层谱系的能力。本研究涉及从人脐带中分离和鉴定间充质干细胞（MSC）及其向功能性胰岛的分化。由于这些 MSC 被发现持续表达巢蛋白，我们假设它们是胰岛新生的理想候选者，无需进一步操作。人脐带基质干细胞（hUCMSCs）表达 CD29、CD44、CD73、CD90、CD105、平滑肌肌动蛋白、巢蛋白、波形蛋白、增殖标志物 Ki67 和胚胎标志物 Oct4、SSEA4。它们被发现 CD33、CD34、CD45 和 HLA-DR 阴性。hUCMSCs 具有长期高增殖能力，表明其具有扩大规模的潜力。当用特定的分化因子混合物处理时，这些细胞分化为脂肪、软骨、骨、神经元和胰岛样簇（ILC）。这些 ILC 对二苯基硫代卡巴腙（DTZ）染色呈阳性，并表达人 C 肽、胰岛素和胰高血糖素。新生成的胰岛的实时 qPCR 分析进一步证明了 Pdx-1、Ngn3、胰岛素、胰高血糖素和生长抑素转录本的丰富性。在实验性糖尿病小鼠移植中，这些 ILC 恢复了正常血糖水平、体重，并表现出正常的葡萄糖耐量试验，表明其功能状态。因此，本研究表明，脐带中持续表达巢蛋白的阳性祖细胞作为胰岛新生的一种新来源具有潜力，并可用于糖尿病的细胞替代治疗。
