Human Adipose-Derived Mesenchymal Stromal Cells Exhibit High HLA-DR Levels and Altered Cellular Characteristics under a Xeno-free and Serum-free Condition

Dam, Phuong T.M.; Hoang, Van T.; Bui, Hue Thi Hong; Hang, Le Minh; Hoang, Duc M.; Nguyen, Hoang Phuong; Lien, Ha Thi; Tran, Huong Thi Thanh; Nguyen, Xuan Hung; Nguyen Thanh, Liem

Abstract

Background: We have observed an increased expression of negative markers in some clinical-grade, xeno- and serum-free cultured adipose-derived mesenchymal stem/stromal cell (ADMSC) samples. It gave rise to concern that xeno- and serum-free conditions might have unexpected effects on human ADMSCs. This study aims to test this hypothesis for two xeno- and serum-free media, Power Stem MSC1 media (PS) and Stem MACS MSC Expansion Media (SM), that support the in vitro expansion of ADMSCs.

Methods: We investigated the expression of negative markers in 42 clinical-grade ADMSC samples expanded in PS. Next, we cultured ADMSCs from seven donors in PS and SM and examined their growth and colony-forming ability, surface marker expression, differentiation, cell cycle and senescence, as well as genetic stability of two passages representing an early and late passage for therapeutic MSCs.

Results: 15 of 42 clinical-grade PS-expanded ADMSC samples showed an increased expression of negative markers ranging from 2.73% to 34.24%, which positively correlated with the age of donors. This rise of negative markers was related to an upregulation of Human Leukocyte Antigen – DR (HLA-DR). In addition, the PS-cultured cells presented decreased growth ability, lower frequencies of cells in S/G2/M phases, and increased ß-galactosidase activity in passage 7 suggesting their senescent feature compared to those grown in SM. Although MSCs of both PS and SM cultures were capable of multilineage differentiation, the PS-cultured cells demonstrated chromosomal abnormalities in passage 7 compared to the normal karyotype of their SM counterparts.

Conclusions: These findings suggest that the SM media is more suitable for the expansion of therapeutic ADMSCs than PS. The study also hints a change of ADMSC features at more advanced passages and with increased donor’s age. Thus, it emphasizes the necessity to cover these aspects in the quality control of therapeutic MSC products.

Citation

Dam, P. T., Hoang, V. T., Bui, H. T. H., Hang, L. M., Hoang, D. M., Nguyen, H. P., Lien, H. T., Tran, H. T. T., Nguyen, X. H., & Nguyen Thanh, L. (2021). Human Adipose-Derived Mesenchymal Stromal Cells Exhibit High HLA-DR Levels and Altered Cellular Characteristics under a Xeno-free and Serum-free Condition. Stem Cell Reviews and Reports, 17(6), 2291-2303. https://doi.org/10.1007/s12015-021-10242-7

• Journal Article Type: Article
• Acceptance Date: Aug 11, 2021
• Online Publication Date: Sep 11, 2021
• Publication Date: Dec 1, 2021
• Deposit Date: Oct 19, 2024
• Publicly Available Date: Oct 22, 2024
• Journal: Stem Cell Reviews and Reports
• Print ISSN: 1550-8943
• Electronic ISSN: 1558-6804
• Publisher: Springer Verlag
• Peer Reviewed: Peer Reviewed
• Volume: 17
• Issue: 6
• Pages: 2291-2303
• DOI: https://doi.org/10.1007/s12015-021-10242-7
• Keywords: Adipose-derived stem cells, Adipose-derived mesenchymal stem cells, Adipose-derived mesenchymal stromal cells, HLA-DR, Karyotype, Senescence
• Public URL: https://nottingham-repository.worktribe.com/output/40563066
• Publisher URL: https://link.springer.com/article/10.1007/s12015-021-10242-7
• Additional Information: Accepted: 11 August 2021; First Online: 11 September 2021; : ; : Sample collection and data analysis were approved by the Ethics Committee of Vinmec Healthcare System and were carried out in accordance with the Declaration of Helsinki. Informed consent was signed by all donors before any study-related interventions took place.; : Not applicable; : The authors of the study are employed by the not-for-profit Vinmec Healthcare System.