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Abstract

Background Mature adipocytes are difficult to study ex vivo, prompting the use of human adipose progenitor cells (hAPCs). However, hAPCs undergo replicative senescence, limiting their utility in long-term studies. Methods We inserted human telomerase reverse transcriptase (TERT) into the AAVS1 locus of CD55+ hAPCs derived from abdominal subcutaneous adipose tissue, and characterized the cells before and after adipogenic differentiation. Results TERT-hAPCs retained proliferative and adipogenic capacities for over 80 passages, comparable to early-passage wild type hAPCs. Transcriptomic and proteomic analyses confirmed strong adipocyte gene expression. Functionally, TERT-hAPCs responded to insulin and lipolytic stimuli (isoprenaline, dibutyryl cAMP, TNF-α). They adapted well to both 2D and 3D cultures, with improved adipogenesis under spheroid conditions. Conclusion Immortalization of CD55+ hAPCs yields cells with stable proliferative and adipogenic capacity across passages. Being cryopreservable and suitable for both 2D and 3D cultures, TERT-hAPCs offer a reliable, reusable model system for adipocyte studies using cells with a consistent genetic background.

Keywords

Genetic engineering; CRISPR/Cas9; multiomics; spheroid; fat cell; adipocyte; cell model; adipogenesis; lipolysis

Published in

Adipocyte
2025, volume: 14, number: 1, article number: 2283213

SLU Authors

UKÄ Subject classification

Cell and Molecular Biology

Publication identifier

  • DOI: https://doi.org/10.1080/21623945.2023.2283213

Permanent link to this page (URI)

https://res.slu.se/id/publ/142064