目的利用流式细胞仪分选人脂肪干细胞(Human adipose-derived stem cells,hADSCs)并检测其体外成骨活性。方法分离hADSCs,通过流式细胞仪以CD105作为表面标志进行分选,所得细胞成骨诱导培养,以CD105-细胞、未分选细胞作为对照。2周后进行碱性磷酸酶(AKP)和骨钙蛋白(OCN)定量PCR和Western-blot检测。结果流式细胞分选所得CD105+hADSCs细胞约占单核细胞的50%。定量PCR和Western-blot检测均显示分选hADSCs成骨诱导后,AKP和OCN的表达均显著高于CD105-细胞组及未分选细胞组(P<0.05)。结论诱导分选纯化后的hADSCs有较好的成骨活性,可作为骨组织工程的种子细胞。

Objective To explore the osteogenesis ability of purified hADSCs through flow cytometry sorting in vitro.Methods hADSCs were isolated and separated through flow cytometry using CD105 as surface marker.These cells were cultured in osteogenic-induced medium,CD105-hADSCs and non-seperated hADSCs were served as control groups.The cells were examined for alkaline phosphatase(AKP) and osteocalcin(OCN) by real-time PCR and western-blot after 2 weeks culture.Results The percentage of CD105+ hADSCs in monocytes was about 50% through flow cytometry sorting.Real-time PCR and western-blot both revealed the AKP and OCN activity of CD105+ hADSCs were significantly higher than CD105-hADSCs and non-seperated hADSCs after 2 weeks osteogenic-induction(P<0.05).Conclusion The hADSCs purified have good osteogenesis ability and could be the seeding cells for bone engineering.