Kadam Sachin, Muthyala Sudhakar, Nair Prabha, Bhonde Ramesh
National Center for Cell Science, Ganeshkhind, Pune 411007, MS, India.
Rev Diabet Stud. 2010 Summer;7(2):168-82. doi: 10.1900/RDS.2010.7.168. Epub 2010 Aug 10.
Placental tissue holds great promise as a source of cells for regenerative medicine due to its plasticity, and easy availability. Human placenta-derived mesenchymal stem cells (hPDMSCs) have the potential to differentiate into insulin-producing cells. Upon transplantation, they can reverse experimental diabetes in mice. However, it is not known whether culture-expanded undifferentiated hPDMSCs are capable of restoring normoglycemia upon transplantation in streptozotocin (STZ)-induced diabetic mice. Hence we prepared long-term cultures of hPDMSCs from the chorionic villi of full-term human placenta. Flow cytometry analyses and immunocytochemistry study revealed bonafide mesenchymal nature of the isolated hPDMSCs. These cultures could differentiate into adipogenic, oesteogenic, chondrogenic, and neuronal lineages on exposure to lineage-specific cocktails. Furthermore, we showed that hPDMSCs can form islet-like cell clusters (ILCs) on stepwise exposure to serum-free defined media containing specific growth factors and differentiating agents. qRT-PCR showed the expression of insulin, glucagon, and somatostatin in undifferentiated hPDMSCs and in ILCs. Differentiated ILCs were found to express human insulin, glucagon, and somatostatin by immunocytochemistry. Additionally, ILCs also showed abundance of pancreatic transcription factors ngn3 and isl1. Both undifferentiated hPDMSCs and ILCs exihibited insulin secretion in response to glucose. Transplantation of hPDMSCs or ILCs derived from hPDMSCs in STZ-induced diabetic mice led to restoration of normoglycemia. Our results demonstrate, for the first time, reversal of hyperglycemia by undifferentiated hPDMSCs and ILCs derived from hPDMSCs. These results suggest human placenta-derived MSCs as an alternative source for cell replacement therapy in diabetes.
胎盘组织因其可塑性和易于获取，作为再生医学的细胞来源具有巨大潜力。人胎盘来源的间充质干细胞（hPDMSCs）有分化为胰岛素分泌细胞的潜力。移植后，它们可逆转小鼠的实验性糖尿病。然而，尚不清楚培养扩增的未分化hPDMSCs在链脲佐菌素（STZ）诱导的糖尿病小鼠中移植后是否能够恢复正常血糖水平。因此，我们从足月人胎盘的绒毛膜绒毛中制备了hPDMSCs的长期培养物。流式细胞术分析和免疫细胞化学研究揭示了分离出的hPDMSCs具有真正的间充质特性。这些培养物在接触特定谱系的混合试剂后可分化为脂肪生成、成骨、软骨生成和神经谱系。此外，我们表明，hPDMSCs在逐步接触含有特定生长因子和分化剂的无血清限定培养基时可形成胰岛样细胞簇（ILCs）。qRT-PCR显示未分化的hPDMSCs和ILCs中存在胰岛素、胰高血糖素和生长抑素的表达。通过免疫细胞化学发现分化的ILCs表达人胰岛素、胰高血糖素和生长抑素。此外，ILCs还显示出丰富的胰腺转录因子ngn3和isl1。未分化的hPDMSCs和ILCs在对葡萄糖的反应中均表现出胰岛素分泌。将hPDMSCs或源自hPDMSCs的ILCs移植到STZ诱导的糖尿病小鼠中可导致正常血糖水平的恢复。我们的结果首次证明未分化的hPDMSCs和源自hPDMSCs的ILCs可逆转高血糖。这些结果表明人胎盘来源的间充质干细胞可作为糖尿病细胞替代治疗的替代来源。
