Differentiation Potential of Nestin (+) and Nestin (-) Cells Derived from Human Bone Marrow Mesenchymal Stem Cells into Functional Insulin Producing Cells

نویسندگان

  • Abdel-Aziz Abdel-Aziz Biochemistry Division, Chemistry Department, Faculty of Science, Mansoura University, Mansoura, Egypt.
  • Ali Fouad Department of Biotechnology, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
  • Amani Ismail Department of Biotechnology, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
  • Ayman Refaie Nephrology Department, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
  • Mahmoud Gabr Department of Biotechnology, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
  • Mahmoud Zakaria Department of Biotechnology, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
  • Sahar Rashed Department of Biotechnology, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
  • Sherry Khater Department of Biotechnology, Urology and Nephrology Center, Mansoura University, Mansoura, Egypt.
چکیده

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.

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عنوان ژورنال:

دوره 8  شماره 1

صفحات  0- 0

تاریخ انتشار 2019-05

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