Department of Pediatric Endocrinology and Metabolism, Mofid Children's Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Zabol University of Medical Sciences, Zabol, Iran.
Mol Biol Rep. 2022 May;49(5):3539-3548. doi: 10.1007/s11033-022-07194-7. Epub 2022 Feb 2.
Mesenchymal stem cells (MSCs) from human adipose tissue and bone marrow have a great potential for use in cell therapy due to their ease of isolation, expansion, and differentiation. Our intention was to isolate and promote in vitro expansion and differentiation of MSCs from human adipose and bone marrow tissue into cells with a pancreatic endocrine phenotype and to compare the potency of these cells together.
MSCs were pre-induced with nicotinamide, mercaptoethanol, B-27 and b-FGF in L-DMEM for 2 days and re-induced again in supplemented H-DMEM for another 3 days. Expression of five genes in differentiated beta cells was evaluated by Real-time PCR and western blotting and the potency of insulin release in response to glucose stimulation was evaluated by insulin and C-peptide ELISA kit. The differentiated cells were evaluated by immunocytochemistry staining for Insulin and PDX-1. Quantitative RT-PCR results showed up-regulation of four genes in differentiated beta-islet cells (Insulin, Ngn-3, Pax-4 and Pdx-1) compared with the control. Western blot analysis showed that MSCs cells mainly produced proinsulin and insulin after differentiation but nestin was more expressed in pre-differentiated stem cells. Glucose and insulin secretion assay showed that insulin levels and C-peptide secretion were significantly increased in response to 10 mM glucose.
Our study showed that both adipose and bone marrow stem cells could differentiate into functional beta-islet cells but it seems that adipose stem cells could be a better choice for treatment of diabetes mellitus according to their higher potency.
由于易于分离、扩增和分化，源自人脂肪组织和骨髓的间充质干细胞（MSCs）在细胞治疗中具有巨大的应用潜力。我们的目的是分离并促进人脂肪和骨髓组织中的 MSC 在体外扩增和分化为具有胰腺内分泌表型的细胞，并比较这些细胞的潜力。
MSCs 在 L-DMEM 中先用烟酰胺、巯基乙醇、B-27 和 b-FGF 预诱导 2 天，然后在补充 H-DMEM 中再次诱导 3 天。通过实时 PCR 和 Western blot 评估分化的β细胞中五个基因的表达，并通过胰岛素和 C-肽 ELISA 试剂盒评估对葡萄糖刺激的胰岛素释放的效力。通过胰岛素和 PDX-1 的免疫细胞化学染色评估分化细胞。定量 RT-PCR 结果显示，与对照组相比，分化的β胰岛细胞（胰岛素、Ngn-3、Pax-4 和 Pdx-1）中四个基因的表达上调。Western blot 分析表明，MSCs 细胞在分化后主要产生胰岛素原和胰岛素，但巢蛋白在预分化的干细胞中表达更多。葡萄糖和胰岛素分泌测定表明，胰岛素水平和 C-肽分泌在 10mM 葡萄糖刺激下显著增加。
我们的研究表明，脂肪和骨髓干细胞都可以分化为功能性β胰岛细胞，但根据其更高的效力，脂肪干细胞似乎是治疗糖尿病的更好选择。
