不同个体来源和低氧条件对人骨髓间充质干细胞生物学特性的影响

Effects of different donor sources and hypoxia conditions on the biological characteristics of human bone marrow-derived mesenchymal stem cells

目的研究6株不同个体(男:M1、M2、M3;女:F1、F2、F3)来源人骨髓间充质干细胞(BM-MSC)在不同低氧条件下生物学特性的差异。方法流式及诱导分化实验鉴定间充质干细胞的标志物表达及三系分化能力。Transwell法检测迁移能力。CCK8法检测5%及1%氧气下细胞活力差异。结果6株BM-MSC阳性标志物表达均高于99%,阴性标志物总表达均低于2%,且均具有三系分化能力。不同个体来源BM-MSC成骨、成脂分化效率和细胞活力具有差异,性别分层统计显示成脂分化差异可能与性别相关;不同低氧条件下BM-MSC的细胞活力和细胞迁移能力具有差异,5%氧气条件显著优于1%氧气条件。同时考虑氧气和性别因素时,发现不同低氧条件下男性来源的BM-MSC活力相对女性均具有较高的趋势,而迁移能力则是女性来源的BM-MSC相对更强。结论提示应根据人骨髓间充质干细胞的应用目的所需特性对供者个体和扩增环境设立特定标准。

Objective To investigate the differences in biological properties of human bone marrow-derived mesenchymal stem cells(BM-MSCs)from six different individual donors(male:M1,M2,M3;female:F1,F2,F3)under different low oxygen concentration.Methods Flow cytometry and in vitro differentiation assays were performed to identify the biomarker expressions and tri-lineage differentiation abilities of MSCs.Transwell method was performed to assess cell migration ability.Cell viability under 5%and 1%oxygen were measured by CCK8 test.Results All six BM-MSC strains showed a high expression level(above 99%)for positive markers and a low expression level(below 2%)for negative markers,and had the ability to differentiate into three lineages.There are differences in the osteogenic and adipogenic differentiation efficiencies as well as cell viability of BM-MSC derived from different donors.And the differences in adipogenic differentiation may be associated with gender.Both cell viability and migration ability of BM-MSC showed variations between different oxygen conditions,with 5%oxygen conditions showing significantly better outcomes than 1%oxygen conditions,When both oxygen and gender factors were considered,it was found that BM-MSC derived from male donors showed a greater tendency for higher viability under various hypoxic conditions,whereas BM-MSC derived from female donors exhibited relatively stronger migration ability.Conclusion The study suggests that specific standards should be established for the individual donor and the expansion conditions based on the desired characteristics for BM-MSC applications.