Effect assessment of a type of xeno-free and serum-free human adipose-derived mesenchymal stem cells culture medium by proliferation and differentiation capacities

Shanshan Chen; Li Meng; Shanshan Wang; Yan Xu; Wenbin Chen; Jianfeng Wei

doi:10.1007/s10616-023-00586-y

Effect assessment of a type of xeno-free and serum-free human adipose-derived mesenchymal stem cells culture medium by proliferation and differentiation capacities

Cytotechnology. 2023 Oct;75(5):403-420. doi: 10.1007/s10616-023-00586-y. Epub 2023 Jul 11.

Authors

Shanshan Chen^{1

2}, Li Meng¹, Shanshan Wang², Yan Xu¹, Wenbin Chen^{3

4}, Jianfeng Wei^{1

2}

Affiliations

¹ Jiangsu Key Laboratory of Brain Disease Bioinformation, Research Center for Biochemistry and Molecular Biology, Xuzhou Medical University, Xuzhou, China.
² Department of Histology and Embryology, School of Basic Medical Sciences, Xuzhou Medical University, Xuzhou, China.
³ Department of Plastic Surgery, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁴ Department of Burns and Plastic Surgery, The Affiliated Huai'an Hospital of Xuzhou Medical University and the Second People's Hospital of Huai'an City, Huai'an, China.

Abstract

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.

Keywords: Cell culture medium; Mesenchymal stem cells; Serum-free; Xeno-free.

© The Author(s), under exclusive licence to Springer Nature B.V. 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.