Rashed Sahar, Gabr Mahmoud, Abdel-Aziz Abdel-Aziz, Zakaria Mahmoud, Khater Sherry, Ismail Amani, Fouad Ali, Refaie Ayman
Department of Biotechnology, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
Biochemistry Division, Chemistry Department, Faculty of Science, Mansoura University, Mansoura, Egypt.
Int J Mol Cell Med. 2019 Winter;8(1):1-13. doi: 10.22088/IJMCM.BUMS.8.1.1. Epub 2019 Jul 17.
The feasibility of isolating and manipulating mesenchymal stem cells (MSCs) from human patients provides hope for curing numerous diseases and disorders. Recent phenotypic analysis has shown heterogeneity of MSCs. Nestin progenitor cell is a subpopulation within MSCs which plays a role in pancreas regeneration during embryogenesis. This study aimed to separate nestin cells from human bone marrow MSCs, and differentiate these cells into functional insulin producing cells (IPCs) compared with nestin cells. Manual magnetic separation was performed to obtain nestin cells from MSCs. Approximately 91±3.3% of nestin cells were positive for anti-nestin antibody. Pluripotent genes were overexpressed in nestin cells compared with nestin cells as revealed by quantitative real time-PCR (qRT-PCR). Following differentiation, flow cytometric analysis showed that 2.7±0.5% of differentiated nestin cells were positive for anti-insulin antibody in comparison with 0.08±0.02% of nestin cells. QRT-PCR showed higher expression of insulin and other endocrine genes in comparison with nestin cells. While immunofluorescence technique showed the presence of insulin and C-peptide granules in nestin cells. Therefore, our results introduced nestin cells as a pluripotent subpopulation within human MSCs which is capable to differentiate and produce functional IPCs.
从人类患者中分离和操纵间充质干细胞（MSCs）的可行性为治愈众多疾病和病症带来了希望。最近的表型分析显示了MSCs的异质性。巢蛋白祖细胞是MSCs中的一个亚群，在胚胎发育过程中的胰腺再生中发挥作用。本研究旨在从人骨髓MSCs中分离巢蛋白⁺细胞，并将这些细胞与巢蛋白⁻细胞相比分化为功能性胰岛素产生细胞（IPCs）。进行手动磁性分离以从MSCs中获得巢蛋白⁺细胞。约91±3.3%的巢蛋白⁺细胞抗巢蛋白抗体呈阳性。定量实时PCR（qRT-PCR）显示，与巢蛋白⁻细胞相比，巢蛋白⁺细胞中多能基因过度表达。分化后，流式细胞术分析显示，分化的巢蛋白⁺细胞中有2.7±0.5%抗胰岛素抗体呈阳性，而巢蛋白⁻细胞为0.08±0.02%。qRT-PCR显示，与巢蛋白⁻细胞相比，巢蛋白⁺细胞中胰岛素和其他内分泌基因的表达更高。免疫荧光技术显示巢蛋白⁺细胞中存在胰岛素和C肽颗粒。因此，我们的结果表明巢蛋白⁺细胞是人类MSCs中的一个多能亚群，能够分化并产生功能性IPCs。
