In Vitro Differentiation of Insulin Secreting Cells
from Mouse Bone Marrow Derived Stage-Specific
Embryonic Antigen 1 Positive Stem Cells
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 mar- row 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 characteriza- tion 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.
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 immunocytochem-
istry 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
Our study clearly demonstrates the potential of SSEA-1 positive cells to differentiate into insulin secreting cells in defined culture conditions for clinical ap- plications.