Chen Shanshan, Meng Li, Wang Shanshan, Xu Yan, Chen Wenbin, Wei Jianfeng
Jiangsu Key Laboratory of Brain Disease Bioinformation, Research Center for Biochemistry and Molecular Biology, Xuzhou Medical University, Xuzhou, China.
Department of Histology and Embryology, School of Basic Medical Sciences, Xuzhou Medical University, Xuzhou, China.
Cytotechnology. 2023 Oct;75(5):403-420. doi: 10.1007/s10616-023-00586-y. Epub 2023 Jul 11.
Human mesenchymal stem cells (hMSCs) possess broad prospects in pre-clinical research. In vitro amplification of hMSCs requires appropriate medium to reach the number of seed cells with clinical significance. However, the uncertainty of the heterologous components of the traditional fetal bovine serum (FBS) culture medium has great safety risks. Moreover, existing commercial hMSCs medium is very expensive, therefore a safer and more optimal hMSCs medium is urgently needed. Accordingly, we developed five components adipose-derived hMSCs (hADMSCs) medium without xenogenic components, named E5 SFM. which is mainly composed of knockout serum replacement (KSR), and additionally four components such as fibroblast growth factor and transferrin. Here, we mainly compared the E5 SFM with traditional FBS-containing medium and a commercial medium by surface markers testing, proliferation assay as well as osteogenic, adipogenic and chondrogenic differentiation assessment. We demonstrated that hADMSCs cultured in the E5 SFM showed similar morphological characteristics and immunophenotypes to those in other media. Notably, cell proliferative capability was similar to that in the commercial medium, but higher than that in the FBS-containing medium and other media. Additionally, their capabilities of adipogenic and osteogenic differentiation were significantly higher than those of other media. Consequently, we concluded that the E5 SFM medium can not only effectively promote cell proliferation of hMSCs, but also has optimal differentiative capacity and clear and simple ingredients.
人间充质干细胞(hMSCs)在临床前研究中具有广阔前景。hMSCs的体外扩增需要合适的培养基以获得具有临床意义的种子细胞数量。然而,传统胎牛血清(FBS)培养基中异源成分的不确定性存在很大安全风险。此外,现有的商业化hMSCs培养基非常昂贵,因此迫切需要一种更安全、更优化的hMSCs培养基。据此,我们开发了一种不含异种成分的五成分脂肪来源的hMSCs(hADMSCs)培养基,命名为E5 SFM。它主要由敲除血清替代品(KSR)组成,另外还有成纤维细胞生长因子和转铁蛋白等四种成分。在此,我们主要通过表面标志物检测、增殖测定以及成骨、成脂和成软骨分化评估,将E5 SFM与传统含FBS培养基和一种商业化培养基进行比较。我们证明,在E5 SFM中培养的hADMSCs与在其他培养基中培养的hADMSCs具有相似的形态特征和免疫表型。值得注意的是,其细胞增殖能力与商业化培养基中的相似,但高于含FBS培养基和其他培养基中的。此外,它们的成脂和成骨分化能力明显高于其他培养基。因此,我们得出结论,E5 SFM培养基不仅能有效促进hMSCs的细胞增殖,而且具有最佳的分化能力,成分清晰简单。