人血小板裂解液对骨髓间质干细胞成骨成脂分化的影响

目的探讨人血小板裂解液(HPL)体外培养人骨髓间质干细胞(MSCs)及其对MSCs成骨和成脂分化能力的影响。方法采用贴壁法分离培养3株人骨髓来源MSCs,每株细胞在培养时平行分为2个实验组,分别用含10%胎牛血清的低糖培养基和含7.5%HPL的低糖培养基培养,均培养MSCs至第5代,分别用成骨及成脂诱导液对每组MSCs做成骨、成脂分化诱导,诱导成功后分别用茜素红、油红O对成骨和成脂分化结果染色鉴定,并进一步对成骨、成脂诱导结果做定量分析:成骨分化染色后洗脱茜素红在562nm波长下测定OD值并对每个培养孔细胞做蛋白定量,以茜素红OD值与蛋白定量值的比值作为成骨分化的定量值;成脂分化染色后用异丙醇洗脱油红O,在510nm波长下测定OD值,以之作为成脂分化的定量值。对所得的结果做统计学分析。结果对第5代的MSCs做成骨诱导12d后,光镜下可见致密结节形成,茜素红染色及定量鉴定结果显示2种培养液培养的MSCs均能成功向成骨细胞分化,而7.5%HPL培养的MSCs成骨能力比10%FBS培养的MSCs强,定量鉴定结果分别为16.548±4.397、4.151±5.631(P<0.05)。成脂诱导12d后,细胞内有明显脂滴形成,油红O染色及定量分析结果显示MSCs成功向脂肪细胞分化,但7.5%HPL培养的MSCs成脂能力与10%FBS培养的MSCs相比较弱,定量鉴定结果分别为0.239±0.030、0.497±0.105(P<0.05)。结论 HPL可以促进MSCs成骨分化,抑制其成脂分化。

Effect of human platelet lysate on the multiple differentiation capacities of bone marrow mesenchymal stem cells

Objective To investigate the effect of human platelet lysate(HPL) on the multiple differentiation capacities of mesenchymal stem cells(MSCs) from the bone marrow,and to provide guidance for the applications of MSCs in tissue engineering and certain diseases.Methods MSCs were isolated from the bone marrow of 3 healthy donors,and then were cultured in 10%FBS or 7.5%HPL for induction of osteogenic or adipogenic cells using the cultured cells at the 5th passage.Osteogenic and adipogenic differentiation potential were analyzed by alizarin red S staining or Oil red O staining,and quantitative determination.Results We found that MSCs cultured in both 10%FBS and 7.5%HPL media could be induced successfully into osteogenic or adipogenic cells.The quantitative data(16.548±4.397 vs 4.151±5.631) showed that MSCs cultured in HPL media had,in comparison to MSCs pre-cultured with FBS,a higher potential for osteogenic differentiation(P<0.05) .On the contrary,the adipogenic differentiation of MSCs cultured in HPL media were reduced(0.239±0.030 vs 0.497±0.105,P<0.05) .Conclusion Based on the results of the present study,HPL enhances the osteogenic differentiation of MSCs,while impairing the adipogenic differentiation.