【摘 要】
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Background: Recent studies have shown that induced pluripotent stem cells (iPSCs) could be maintained in an undifferentiated state and subsequently be differentiated into mesenchymal stem cells (iPSC-
【机 构】
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Department of Bioengineering, College of Chemistry, Chemical Engineering and Biotechnology,Donghua U
【出 处】
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第十一届全国再生医学(干细胞与组织工程)学术研讨会暨第七届全国组织工程与再生医学大会
论文部分内容阅读
Background: Recent studies have shown that induced pluripotent stem cells (iPSCs) could be maintained in an undifferentiated state and subsequently be differentiated into mesenchymal stem cells (iPSC-MSCs) with notable advantages over iPSCs per se.To regulate iPSC-MSCs specific differentiation for bone tissue engineering, the present study was purposefully designed to explore the efficacy of osteoblast-conditioned medium (CM) in directing mouse iPSC-MSCs differentiation into osteogenic lineage.Methods: Osteoblasts were harvested from one-day-old ICR mice cranium by adherent culture and enzyme digestion methods.Osteogenic differentiation medium was prepared by mixing the CM with basic medium (BM) at ratio 3∶7, 5∶5 and 7∶3, respectively.iPSC-MSCs were derived from mouse iPSCs by treatment with retinoic acid (RA) during later embryonic bodies formation.Different concentrations of CM were administered to iPSC-MSCs culture throughout 14 days.At predefined time points, iPSC-MSC-induced cells were evaluated by alkaline phosphatase activity (ALP), quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and immunohistochemistry staining.Results: Osteoblast CM at all concentrations distinctly promoted iPSC-MSCs to become poly-edged morphologically after induction for 14 days.Histological analysis revealed the characteristics of osteogenic differentiation with strong staining for ALP, immunochemical expression of osteogenic marker Runx2, and enhanced expressions of ? osteogenesis-related genes, alp, runx2, col and ocn mRNA.The most effective CM concentration was identified to be 30%.Conclusions: Defined mouse osteoblast conditioned culture medium can significantly induce the differentiation of iPS-MSCs towards osteoblasts, which may serve as the potential source of osteogenic seed cells for bone tissue engineering.
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