Sun Yu, Zhang Mengchao, Ji Shangwei, Liu Lin
Department of Intervention, China‑Japan Union Hospital of Jilin University, Changchun, Jilin 130033, P.R. China.
Department of Radiology, China‑Japan Union Hospital of Jilin University, Changchun, Jilin 130033, P.R. China.
Mol Med Rep. 2015 Nov;12(5):6835-40. doi: 10.3892/mmr.2015.4305. Epub 2015 Sep 9.
Mesenchymal stem cells (MSCs) with the ability to differentiate into insulin‑producing cells (IPCs) have become the most promising means of therapy for diabetes mellitus. Adipose‑derived stromal cells (AdSCs), having similar characteristics to those of derived MSCs, are known to exhibit extensive proliferation potential and are able to undergo multi‑lineage differentiation. Whether AdSCs can differentiate into insulin‑producing cells (IPCs), however, has not been sufficiently elucidated. Therefore, the present study sought to investigate the in vitro differentiation of rabbit (r)AdSCs into IPCs, which may provide an abundant source of cells to treat diabetes. rADSCs were obtained from liposuction aspirates and then induced with glucagon‑like peptide‑1 and nicotinamide to differentiate into insulin‑secreting cells. Differentiation was evaluated by the analysis of morphology, dithizone (DTZ) staining, reverse transcription polymerase chain reaction (RT‑PCR), western blot analysis and a glucose challenge assay with detection of insulin secretion by ELISA. Morphological phase‑contrast microscopic observation revealed typical islet‑like cell clusters following 21 days of differentiation. DTZ staining also showed that differentiated cells were positive and undifferentiated cells were negative for insulin production. Furthermore, RT‑PCR analysis confirmed the mRNA expression of insulin, PDX1 and GLUT2 in differentiated cells. Western blot analysis showed that insulin was expressed by the differentiated cells. The glucose challenge assay showed that insulin secretion of the IPCs was in a glucose dependent manner. These findings implied that AdSCs are able to differentiate into IPC in vitro, and are therefore promising candidates for the treatment of diabetes.
具有分化为胰岛素生成细胞(IPC)能力的间充质干细胞(MSC)已成为治疗糖尿病最有前景的手段。脂肪来源的基质细胞(AdSC)与来源的MSC具有相似的特征,已知其具有广泛的增殖潜力,并能够进行多谱系分化。然而,AdSC是否能够分化为胰岛素生成细胞(IPC)尚未得到充分阐明。因此,本研究旨在探讨兔(r)AdSC在体外分化为IPC的情况,这可能为治疗糖尿病提供丰富的细胞来源。rADSC从抽脂抽吸物中获得,然后用胰高血糖素样肽-1和烟酰胺诱导分化为胰岛素分泌细胞。通过形态学分析、双硫腙(DTZ)染色、逆转录聚合酶链反应(RT-PCR)、蛋白质印迹分析以及用ELISA检测胰岛素分泌的葡萄糖激发试验来评估分化情况。形态学相差显微镜观察显示,分化21天后出现典型的胰岛样细胞簇。DTZ染色还表明,分化细胞对胰岛素产生呈阳性,未分化细胞呈阴性。此外,RT-PCR分析证实了分化细胞中胰岛素、PDX1和GLUT2的mRNA表达。蛋白质印迹分析表明分化细胞表达胰岛素。葡萄糖激发试验表明,IPC的胰岛素分泌呈葡萄糖依赖性。这些发现表明,AdSC能够在体外分化为IPC,因此有望成为治疗糖尿病的候选细胞。