Autophagy Is Involved in Mesenchymal Stem Cell Death in Coculture with Chondrocytes
| Autor: | Carlo Alberto Paggi, Yao Fu, Marcel Karperien, Andre J. van Wijnen, Catalina Galeano Garces, Mario Hevesi, Daniela Galeano Garces, Amel Dudakovic, Allan B. Dietz |
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| Přispěvatelé: | TechMed Centre, Developmental BioEngineering |
| Rok vydání: | 2020 |
| Předmět: |
mesenchymal stem cells
autophagy 0303 health sciences Chemistry Mesenchymal stem cell Autophagy UT-Hybrid-D chondrocytes Biomedical Engineering Cell Differentiation Physical Therapy Sports Therapy and Rehabilitation Coculture Techniques In vitro Cell biology 03 medical and health sciences 0302 clinical medicine In vivo pellet 030220 oncology & carcinogenesis Humans Immunology and Allergy coculture Clinical Research papers 030304 developmental biology |
| Zdroj: | Cartilage Cartilage, 13(2_suppl), 969S-979S. SAGE Publications |
| ISSN: | 1947-6043 1947-6035 |
| DOI: | 10.1177/1947603520941227 |
| Popis: | Objective Cartilage formation is stimulated in mixtures of chondrocytes and human adipose–derived mesenchymal stromal cells (MSCs) both in vitro and in vivo. During coculture, human MSCs perish. The goal of this study is to elucidate the mechanism by which adipose tissue–derived MSC cell death occurs in the presence of chondrocytes. Methods Human primary chondrocytes were cocultured with human MSCs derived from 3 donors. The cells were cultured in monoculture or coculture (20% chondrocytes and 80% MSCs) in pellets (200,000 cells/pellet) for 7 days in chondrocyte proliferation media in hypoxia (2% O2). RNA sequencing was performed to assess for differences in gene expression between monocultures or coculture. Immune fluorescence assays were performed to determine the presence of caspase-3, LC3B, and P62. Results RNA sequencing revealed significant upregulation of >90 genes in the 3 cocultures when compared with monocultures. STRING analysis showed interconnections between >50 of these genes. Remarkably, 75% of these genes play a role in cell death pathways such as apoptosis and autophagy. Immunofluorescence shows a clear upregulation of the autophagic machinery with no substantial activation of the apoptotic pathway. Conclusion In cocultures of human MSCs with primary chondrocytes, autophagy is involved in the disappearance of MSCs. We propose that this sacrificial cell death may contribute to the trophic effects of MSCs on cartilage formation. |
| Databáze: | OpenAIRE |
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