HUMAN BONE MARROW CONTRIBUTES TO ISLET REGNERATION IN VITRO

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。I型糖尿病的特征是胰腺中产生胰岛素的β-胰岛细胞被破坏，导致高血糖和相关的发病率。为了探讨人骨髓细胞(BMC)体外分化为功能性胰岛样细胞的可能性，并检测胰岛样细胞对糖尿病的治疗作用，我们进行了GFP阳性标记的小鼠BMC与胰腺共培养的初步研究。结果发现，小鼠骨髓基质细胞能够聚集形成胰岛样簇(胰岛素原免疫组织化学鉴定)。这些发现使我们能够提出人类BMC转分化为胰岛素产生细胞的可能性。在这个试验性的方案中，假设人的BMC在某些情况下能够分化为胰岛素产生细胞，导致体内葡萄糖的调节。为了验证这一假设，首先，需要优化人BMC的培养液，使其能够分化为新的功能正常的胰岛素产生细胞，并测试BMC是否能够在体外修复受损的胰岛；其次，将评估胰岛素产生细胞的特异性标志物在基因和蛋白质水平的表达。之后，将胰岛样簇植入非肥胖糖尿病严重联合免疫缺陷(NOD-SCID)小鼠体内，将测试这些骨髓来源的胰岛样簇的植入和功能。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Type I diabetes is characterized by the destruction of insulin-producing beta-islet cells in the pancreas, resulting in hyperglycemia and associated morbidity. To explore the possibility of human bone marrow cells (BMC) in vitro differentiating into functional islet-like cells and to test the diabetic therapeutic potency of islet-like cells, GFP positive labeled mice BMC was co-culture with the pancreas in our preliminary studies. It was found that mice BMC are able to aggregate in forming islet like clusters (identified with proinsulin immunohistochemistry). The findings enable us to propose the possibility of human BMC transdifferentiating into insulin producing cells. In this pilot proposal, it is hypothesized that human BMC is able to differentiate into insulin producing cells under certain circumstances, leading to the regulation of glucose in vivo. To test this hypothesis, first, the optimization of the medium for human BMC allowing the differentiation to new functional insulin producing cells will be necessary and along with testing whether BMC will be able to repair damaged islets in vitro; second, the expression of the specific markers for insulin producing cell in gene and protein levels will be evaluated. Thereafter, implantation of the islet like cluster into a non-obese diabetic severe combined immunodeficient (NOD-SCID) mice will test the engraftment and function of these bone marrow derived islet like producing clusters.