Fetal porcine pancreatic stem cells differentiating into Insulin-producing cells in vitro and their curative effect on diabetic nude mice | 胎猪胰腺干细胞体外诱导分化为胰岛素分泌细胞及其移植治疗裸鼠糖尿病的研究

中国人. 缺乏供体细胞制约着胰岛替代疗法在玉型糖尿病治疗过程中的应用。本研究对实验室分离保存的胎猪胰腺干细胞(fetal porcine pancreatic stem cells, FPPSCs)，体外诱导分化为胰岛素分泌细胞以及移植治疗裸鼠糖尿病的能力进行了深入研究，以探讨该细胞株作为异种供体细胞的潜力。研究结果表明，该细胞体外增殖活力旺盛，其形态及表面抗原表达特征与间充质干细胞极其相似，除表达胰腺干细胞相关标志外，FPPSCs还表达一些胚胎干细胞的相关标志。采用无血清诱导方案诱导2周后，FPPSCs 形成DTZ染色阳性的胰岛样细胞团(islet-like cell cluster, ICC)，该细胞团表达胰岛素、Glut-2 等胰岛素分泌细胞功能相关的蛋白。RT-PCR 结果显示，诱导后，胰腺干细胞相关标志表达减弱，而成熟胰岛素分泌细胞相关标志表达增强。葡萄糖刺激实验结果显示，诱导后，FPPSCs合成、分泌胰岛素和C- 肽的能力显著增强(P0.05)。将诱导2 周后的细胞移植到糖尿病裸鼠腹腔内，可缓解其高血糖状况。结果提示，FPPSCs具有间充质干细胞的特性，体外能够诱导分化为胰岛素分泌细胞，并具有改善糖尿病裸鼠高血糖症状的能力，有望为进一步研究异种胰腺干细胞移植治疗糖尿病提供种子细胞。

英语. Islet replacement therapy is limited by the shortage of donor islet cells. The aim of this article is to study the potential capacity of fetal porcine pancreatic stemcells (FPPSCs) as a xenograft donor cells. FPPSCs have a strong proliferation ability, and the expression of it's surface antigen was similar to the mesenchymal stem cells. It expressed not only the markers of pancreatic stem cells, but also the markers of ES cells. After 2 weeks induction using a serum-free protocol, the DTZ-positive islet-like cell clusters (ICC) were formed, and these ICCs expressed insulin and Glut-2. After induction, the expression of markers related to pancreatic stem cells decreased, while that related to insulin-producing cells increased, and the insulin and C-peptide content synthesized and secreted by FPPSCs increased remarkably(P0.05). The blood glucose level of diabetic nude mice in differentiated cells transplantation group decreased, but quickly reached and maintained at a high level (16.7 mmol/L). These results demonstrated that FPP SCs can be induced to differentiate into insulin-producing cells in vitro，and ameliorate the hyperglycemia of diabetic nude mice. It might provide unlimited resources for islet replacement therapy.