摘要
目的探讨血小板裂解物体外培养、扩增人骨髓基质干细胞的可行性。方法将富含血小板血浆以冻融裂解法处理,培养人骨髓基质干细胞;体系中加入不同浓度的裂解物,MTT法检测细胞体外增殖情况;流式细胞仪分析细胞表型特征,并应用细胞化学染色法观察细胞体外成骨和成脂肪能力。结果血小板裂解物培养的骨髓基质干细胞呈典型的成纤维细胞形态,均一呈现CD14、CD31、CD34、CD45及HLA-DR阴性和CD73、CD90、CD105、CD166阳性,具备体外成骨、成脂分化能力。此外,与经筛选的胎牛血清比较,低浓度血小板裂解物即具有支持骨髓基质干细胞扩增的能力。结论血小板裂解物可替代胎牛血清,用于骨髓基质干细胞的体外扩增。
Objective To investigate the suitability and proliferation capacity of human platelet lysate in the production of bone-marrow stromal cells. Methods Human platelet lysates （PL） were prepared by freezing and thawing the platelet rich plasma. Marrow stromal cells （MSCs） were cultured in media containing PL and their proliferation status was evaluated by MTT assay. Furthermore, cell phenotypic characteristics were analyzed by flow cytometry and the differentiation along adipogenic and osteogenic pathways were assessed by histological staining in vitro. Results MSCs cultured in medium containing platelet lysates displayed a typical fibroblast-like morphology. Flow cytometry showed that they were homogenously negative for CD14, CD31, CD34, CD45 and HLA-DR and positive for CD73, CD90, CD105 and CD166. Differentiation capacity was preserved throughout the long-term culture. Compared with fecal calf sera （FCS） selected from lots, platelet lysates exhibited more potent activities to support the expansion of MSCs. Conclusion It is concluded that human platelet lysates might be an optional substitute to FCS for MSCs proliferation in vitro.
出处
《组织工程与重建外科杂志》
2009年第2期61-64,共4页
Journal of Tissue Engineering and Reconstructive Surgery
基金
国家高技术发展计划项目(863)(2007AA02Z454)
国家自然科学基金(30871018)
关键词
骨髓基质细胞
血小板
细胞培养
扩增
Bone marrow stromal cells
Platelet
Cell culture
Proliferation
