Generation of Mesenchymal Cell Lines Derived from Aged Donors

• Published in: International journal of molecular sciences 2021-10, Vol.22 (19), p.10667
• Main Authors: Piñeiro-Ramil, María, Sanjurjo-Rodríguez, Clara, Rodríguez-Fernández, Silvia, Castro-Viñuelas, Rocío, Hermida-Gómez, Tamara, Blanco-García, Francisco J, Fuentes-Boquete, Isaac, Díaz-Prado, Silvia
• Format: Article
• Language: English
• Subjects: Adipocytes; Aged; Antigen T (large); Antigens; Biomedical materials; bone repair; Cartilage; Cartilage diseases; Cell culture; Cell Culture Techniques; Cell cycle; Cell Differentiation; Cell Line; Cell lines; Cell Proliferation; Cell self-renewal; Cells, Cultured; Differentiation; DNA damage; Gene Expression; Humans; immortalization; Immunohistochemistry; Lipids; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - metabolism; mesenchymal stromal cells; Mesenchyme; Mineralization; Osteoarthritis; Osteogenesis; Phenotypes; Regenerative medicine; RNA-directed DNA polymerase; Senescence; Stains & staining; Stromal cells; Telomerase; Telomerase reverse transcriptase; Tissue Donors; Tissue engineering; Transcription factors; Transduction, Genetic
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms221910667

Mesenchymal stromal cells (MSCs) have the capacity for self-renewal and multi-differentiation, and for this reason they are considered a potential cellular source in regenerative medicine of cartilage and bone. However, research on this field is impaired by the predisposition of primary MSCs to senescence during culture expansion. Therefore, the aim of this study was to generate and characterize immortalized MSC (iMSC) lines from aged donors. Primary MSCs were immortalized by transduction of simian virus 40 large T antigen (SV40LT) and human telomerase reverse transcriptase (hTERT). Proliferation, senescence, phenotype and multi-differentiation potential of the resulting iMSC lines were analyzed. MSCs proliferate faster than primary MSCs, overcome senescence and are phenotypically similar to primary MSCs. Nevertheless, their multi-differentiation potential is unbalanced towards the osteogenic lineage. There are no clear differences between osteoarthritis (OA) and non-OA iMSCs in terms of proliferation, senescence, phenotype or differentiation potential. Primary MSCs obtained from elderly patients can be immortalized by transduction of SV40LT and hTERT. The high osteogenic potential of iMSCs converts them into an excellent cellular source to take part in in vitro models to study bone tissue engineering.