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Comparison of Differentiation of Induced Pluripotent Stem Cells and Bone-Marrow Mesenchymal Stem Cells to Osteoblast: Osteogenesis versus Pluripotency.
Foroutan T
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BACKGROUND: Derivation of induced pluripotent stem cells (iPSCs) from various adult somatic cells through over-expression of pluripotent genes could allow for the unlimited autologous supply in regenerative medicine. On the other hand the generation of various progenitors from bone-marrow mesenchymal stem cells (MSCs) is justly well established.
OBJECTIVE: In this study we compared the expression level of pluripotent genes oct4, c-myc, sox-2, nanog, klf4 and lin28 in iPSCs and MSCs derived from bone marrow. Also the potential of osteogenesis of iPSCs and bone-marrow MSCs were compared.
METHODS: We analyzed the expression level of oct4, sox-2, c-myc, klf4, nanog and lin28 genes in human MSCs derived from iPSCs and MSCs by cell culture and real-time PCR. Also the expression level of osteocalcin and osteopontin in both groups were evaluated.
RESULTS: We found that the expression of osteogenic markers in differentiated iPSCs to osteoblast were higher than bone-marrow MSCs. While the levels of pluripotency genes oct4, c-myc and klf4 in iPSCs were significantly (p<0.05) higher than bone-marrow MSCs, MSCs showed higher expression of sox-2, nanog and lin28 compared with iPSCs (p=NS).
CONCLUSION: It seems that the higher expression of osteopontin and osteocalcin in MSCs compared to iPSCs may be due to other factors (besides pluripotency) required for differentiation of stem cells to osteoblast.
Figure 2. Light microscopy image of alizarin red stained MSCs and iPSCs (b) differentiated into osteoblast
Figure 3. Real-time PCR results of osteocalcin (a) and osteopontin (b) genes in iPSCs and MSCs differentiated into osteoblast. *p<0.05
Figure 4. Comparative real-time PCR analysis of oct4, sox-2, c-myc, nanog, klf4 and lin28 genes expression in MSCs and iPSCs. Expression of oct4 and c-myc and klf4 genes in iPSCs is significantly higher than that in MSCs. A significantly (p<0.05) lower level of sox-2, nanog, and lin28 genes expression was detected in iPSCs compared with MSCs significantly
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