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Transcriptomic Characterization of CDK4/hTERT Immortalized Ovine Satellite Cells

ABSTRACT: CDK4/hTERT immortalized ovine satellite cells were established from primary lamb latissimus dorsi muscle and comprehensively characterized using RNA-seq across three populations: primary lamb satellite cells (PLM), early-passage immortalized cells (EPILM, passage 5), and late-passage immortalized cells (LPILM, passage 30). Differential gene expression and functional enrichment analyses were performed to assess immortalization-driven molecular reprogramming, myogenic competence, safety, and suitability for cultivated meat applications.

ORGANISM(S): Ovis aries

PROVIDER: GSE336565 | GEO | 2026/09/03

REPOSITORIES: GEO

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Project description:The secretome of mesenchymal stromal cells (MSCs) can efficiently stimulate regener-ation and therefore is a tempting remedy for “cell-free cellular therapy”. However, the usage of primary MSC cultures as secretome-producers for translation studies has ob-vious obstacles, including rapid aging of MSC cultures, the need for a large number of verified donors and donor-to-donor variability of secretome content. MSCs immortal-ization allows to overcome those limitations and to obtain secretome-producing cul-tures with prolonged lifetime. However, the efficacy and safety of such secretomes are critical issues, which limit their usage as therapeutic agents. In this study we have tested in large detail how the immortalization of MSC cultures affects the content, bi-ological activity and safety of their secretome. MSCs immortalization via overexpres-sion of human TERT gene does not significantly alter the qualitative and quantitative composition of their secretome or its activity according to the results of proteomic analysis, ELISA, qPCR and functional tests in vitro. Moreover, we have demonstrated that the secretome of immortalized MSCs does not contain detectable amounts of te-lomerase and does not possess any transforming activity. Altogether, our data suggest that immortalized MSC cultures may become a reliable source for obtaining standard-ized active secretome in large-scale quantities for clinical use.The secretome of mesenchymal stromal cells (MSCs) can efficiently stimulate regener-ation and therefore is a tempting remedy for “cell-free cellular therapy”. However, the usage of primary MSC cultures as secretome-producers for translation studies has ob-vious obstacles, including rapid aging of MSC cultures, the need for a large number of verified donors and donor-to-donor variability of secretome content. MSCs immortal-ization allows to overcome those limitations and to obtain secretome-producing cul-tures with prolonged lifetime. However, the efficacy and safety of such secretomes are critical issues, which limit their usage as therapeutic agents. In this study we have tested in large detail how the immortalization of MSC cultures affects the content, bi-ological activity and safety of their secretome. MSCs immortalization via overexpres-sion of human TERT gene does not significantly alter the qualitative and quantitative composition of their secretome or its activity according to the results of proteomic analysis, ELISA, qPCR and functional tests in vitro. Moreover, we have demonstrated that the secretome of immortalized MSCs does not contain detectable amounts of te-lomerase and does not possess any transforming activity. Altogether, our data suggest that immortalized MSC cultures may become a reliable source for obtaining standard-ized active secretome in large-scale quantities for clinical use.

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Transcriptomic Characterization of CDK4/hTERT Immortalized Ovine Satellite Cells

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