Author/Authors :
Abouzaripour، Morteza نويسنده Department of Anatomical Sciences, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran , , Pasbakhsh، Parichehr نويسنده , , Atlasi، Nader نويسنده Department of Anatomical Sciences, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran , , Shahverdi، abdol Hossein نويسنده , , Mahmoudi، Reza نويسنده Cellular and Molecular Research Center, Yasuj University of Medical Sciences, Yasuj, Iran , , Ragerdi Kashani، Iraj نويسنده ,
Abstract :
Objective: Bone marrow has recently been recognized as a novel source of stem cells
for the treatment of wide range of diseases. A number of studies on murine bone marrow
have shown a homogenous population of rare stage-specific embryonic antigen 1
(SSEA-1) positive cells that express markers of pluripotent stem cells. This study focuses
on SSEA-1 positive cells isolated from murine bone marrow in an attempt to differentiate
them into insulin-secreting cells (ISCs) in order to investigate their differentiation potential
for future use in cell therapy.
Materials and Methods: This study is an experimental research. Mouse SSEA-1 positive
cells were isolated by Magnetic-activated cell sorting (MACS) followed by characterization
with flow cytometry. Induced SSEA-1 positive cells were differentiated into ISCs with
specific differentiation media. In order to evaluate differentiation quality and analysis,
dithizone (DTZ) staining was use, followed by reverse transcription polymerase chain
reaction (RT-PCR), immunocytochemistry and insulin secretion assay. Statistical results
were analyzed by one-way ANOVA.
Results: The results achieved in this study reveal that mouse bone marrow contains a
population of SSEA-1 positive cells that expresses pluripotent stem cells markers such as
SSEA-1, octamer-binding transcription factor 4 (OCT-4) detected by immunocytochemistry
and C-X-C chemokine receptor type 4 (CXCR4) and stem cell antigen-1 (SCA-1)
detected by flow cytometric analysis. SSEA-1 positive cells can differentiate into ISCs
cell clusters as evidenced by their DTZ positive staining and expression of genes such
as Pdx1 (pancreatic transcription factors), Ngn3 (endocrine progenitor marker), Insulin1
and Insulin2 (pancreaticB-cell markers). Additionally, our results demonstrate expression
of PDX1 and GLUT2 protein and insulin secretion in response to a glucose challenge in
the differentiated cells.
Conclusion: Our study clearly demonstrates the potential of SSEA-1 positive cells
to differentiate into insulin secreting cells in defined culture conditions for clinical applications.
