Senescence during early differentiation reduced the chondrogenic differentiation capacity of mesenchymal progenitor cells

Chantal Voskamp; Wendy J L M Koevoet; Gerjo J V M Van Osch; Roberto Narcisi

doi:10.3389/fbioe.2023.1241338

Senescence during early differentiation reduced the chondrogenic differentiation capacity of mesenchymal progenitor cells

Front Bioeng Biotechnol. 2023 Aug 7:11:1241338. doi: 10.3389/fbioe.2023.1241338. eCollection 2023.

Authors

Chantal Voskamp¹, Wendy J L M Koevoet², Gerjo J V M Van Osch^{1

2

3}, Roberto Narcisi¹

Affiliations

¹ Department of Orthopaedics and Sports Medicine, University Medical Center Rotterdam, Rotterdam, Netherlands.
² Department of Otorhinolaryngology, University Medical Center Rotterdam, Rotterdam, Netherlands.
³ Department of Biomechanical Engineering, Faculty of Mechanical, Maritime and Materials Engineering, Delft University of Technology, Delft, Netherlands.

Abstract

Introduction: Mesenchymal stromal/progenitor cells (MSCs) are promising for cartilage cell-based therapies due to their chondrogenic differentiation capacity. However, MSCs can become senescent during in vitro expansion, a state characterized by stable cell cycle arrest, metabolic alterations, and substantial changes in the gene expression and secretory profile of the cell. In this study, we aimed to investigate how senescence and the senescence-associated secretory phenotype (SASP) affect chondrogenic differentiation of MSCs. Methods: To study the effect of senescence, we exposed MSCs to gamma irradiation during expansion or during chondrogenic differentiation (the pellet culture). Western blot analysis was used to evaluate MSCs response to the chondrogenic inductor TGF-β. Results: When senescence was induced during expansion or at day 7 of chondrogenic differentiation, we observed a significant reduction in the cartilage matrix. Interestingly, when senescence was induced at day 14 of differentiation, chondrogenesis was not significantly altered. Moreover, exposing chondrogenic pellets to the medium conditioned by senescent pellets had no significant effect on the expression of anabolic or catabolic cartilage markers, suggesting a neglectable paracrine effect of senescence on cartilage generation in our model. Finally, we show that senescent MSCs showed lower phosphorylated SMAD2 levels after TGFβ1 stimulation than control MSCs. Conclusion: Overall, these results suggest that the occurrence of senescence in MSCs during expansion or early differentiation could be detrimental for cartilage tissue engineering.

Keywords: MSC; SASP; TGFβ; cartilage; chondrogenesis; regenerative medicine; senescence; tissue engineering.

Grants and funding

This research was financially supported by the Dutch Arthritis Society (ReumaNederland; 16-1-201) and by a TTW Perspectief grant from NWO (William Hunter Revisited; P15-23). This study is part of the Medical Delta RegMed4D program.