Department of Medicine V, University of Heidelberg, Heidelberg, Germany.
Cytotherapy. 2010 Nov;12(7):888-98. doi: 10.3109/14653249.2010.501788.
Culture medium for mesenchymal stromal cells (MSC) is frequently supplemented with fetal calf serum (FCS). FCS can induce xenogeneic immune reactions, transmit bovine pathogens and has a high lot-to-lot variability that hampers reproducibility of results. Several studies have demonstrated that pooled human platelet lysate (HPL) provides an attractive alternative for FCS. However, little is known about the variation between different platelet lysates.
We compared activities of individual HPL on initial fibroblastoid colony-forming units (CFU-F), proliferation, in vitro differentiation and long-term culture. These data were correlated with chemokine profiles of HPL.
Isolation of MSC with either HPL or FCS resulted in similar CFU-F frequency, colony morphology, immunophenotype and adipogenic differentiation potential. Osteogenic differentiation was even more pronounced in HPL than FCS. There were significant differences in MSC proliferation with different HPL, but it was always higher in comparison with FCS. Cell growth correlated with the concentration of platelet-derived growth factor (PDGF) and there was a moderate association with platelet counts. All HPL facilitated expansion for more than 20 population doublings.
Taken together, reliable long-term expansion was possible with all HPL, although there was some variation in platelet lysates of individual units. Therefore the use of donor recipient-matched or autologous HPL is feasible for therapeutic MSC products.
间充质基质细胞(MSC)的培养基通常添加胎牛血清(FCS)。FCS 会引起异种免疫反应、传播牛病原体,并且批次间差异很大,这会影响结果的可重复性。有几项研究表明,人血小板裂解液(HPL)是 FCS 的一种有吸引力的替代品。然而,对于不同血小板裂解液之间的差异,人们知之甚少。
我们比较了单个 HPL 对初始成纤维细胞集落形成单位(CFU-F)、增殖、体外分化和长期培养的活性。这些数据与 HPL 的趋化因子谱相关。
使用 HPL 或 FCS 分离 MSC 会导致相似的 CFU-F 频率、集落形态、免疫表型和脂肪分化潜能。与 FCS 相比,HPL 中的成骨分化更为明显。不同 HPL 之间 MSC 增殖存在显著差异,但与 FCS 相比总是更高。细胞生长与血小板衍生生长因子(PDGF)的浓度相关,与血小板计数存在中度相关性。所有 HPL 都能进行超过 20 次的群体倍增。
总之,虽然个别单位的血小板裂解液存在一些差异,但所有 HPL 都可以实现可靠的长期扩增。因此,对于治疗性 MSC 产品,使用供体-受者匹配或自体 HPL 是可行的。