:: Volume 8, Issue 1 (Int J Mol Cell Med 2019) ::
Int J Mol Cell Med 2019, 8(1): 1-13 Back to browse issues page
Differentiation Potential of Nestin (+) and Nestin (-) Cells Derived from Human Bone Marrow Mesenchymal Stem Cells into Functional Insulin Producing Cells
Sahar Rashed1, Mahmoud Gabr1, Abdel-Aziz Abdel-Aziz2, Mahmoud Zakaria1, Sherry Khater1, Amani Ismail1, Ali Fouad 3, Ayman Refaie4
1- Department of Biotechnology, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
2- Biochemistry Division, Chemistry Department, Faculty of Science, Mansoura University, Mansoura, Egypt.
3- Department of Biotechnology, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt. , alifouad.b@gmail.com
4- Nephrology Department, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
Abstract:   (775 Views)
The feasibility of isolating and manipulating mesenchymal stem cells (MSCs) from human patients provides hope for curing numerous disease and disorders. Recent phenotypic analysis showed heterogeneity of MSCs. A 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 the anti-nestin antibody. Pluripotent genes were overexpressed in nestin (+) cells compared with nestin (-) cells as revealed by quantitative real time-PCR (qRT-PCR). Following in vitro 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 the 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.
Keywords: Human bone marrow derived mesenchymal stem cells, insulin producing cells, real time-quantitative PCR, mesenchymal stem cells, diabetes mellitus
Full-Text [PDF 366 kb]   (278 Downloads)    
Type of Study: Original Article | Subject: Stem Cell
Received: 2019/04/24 | Accepted: 2019/07/13 | Published: 2019/12/6



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