猪脂肪干细胞体外多向分化潜能的实验研究

Hetero-Differentiation of Porcine Adipose-Derived Stem Cells in Vitro

目的探讨猪脂肪干细胞(Adipose-Derived Stem Cells,ADSCs)在体外是否具有多向分化潜能。方法从猪项背部皮下脂肪组织中获取ADSCs,经过体外培养扩增至第2代分别进行成软骨细胞、成骨细胞和成脂肪细胞诱导分化,通过免疫荧光和RT-PCR对其相关表型进行检测。结果猪ADSCs在体外成软骨诱导分化2周后有特异软骨基质Ⅱ型胶原和Aggrecan表达,RT-PCR检测显示有Ⅱ型胶原、Aggrecan和Sox-9mRNA的表达;体外成骨诱导分化2周后有与成骨相关的OPN、OCN和ONN表达;体外成脂肪诱导2周后,经油红染色显示细胞内有脂滴形成。结论猪ADSCs在体外具有向成软骨细胞、成骨细胞和成脂肪细胞分化的潜能。

Objective In this study we investigated whether the porcine adipose-derived stem cells （ADSCs） have muhipotential in vitro. Methods ADSCs were harvested from porcine back adipose tissue and cultured in vitro. Cells of passage 2 were cultured in osteogenic, chondrogenic and adipogenic respectively for two weeks. Muhipotential differentiation phenotype of ADSCs was evaluated by immunofluorescence methods and RT-PCR analysis. Results Expression of Collagen Ⅱ, Aggrecan and Sox-9 were observed in micromass culture that was induced in chondrogenic medium for two weeks; Expression of OPN, OCN, and ONN were observed that was induced in osteogenic medium for two weeks; ADSCs induced in vitro in adipogenic medium after two weeks, Oil red staining showed inter cellular oil droplet deposition. Conclusion The porcine ADSCs can be induced to osteogenic, chondrogenic as well as adipogenic cells.