Couchet, M.* ; Gao, H.* ; Klingelhuber, F. ; Jalkanen, J.* ; De Castro Barbosa, T.* ; Omar-Hmeadi, M.* ; Massier, L.* ; Krahmer, N. ; Mejhert, N.* ; Rydén, M.*
Adipogenic characterization of immortalized CD55+ progenitor cells from human white adipose tissue.
Adipocyte 14:2283213 (2023)
BACKGROUND: Mature adipocytes are notoriously difficult to study ex vivo and alternative cell culture systems have therefore been developed. One of the most common models are human adipose progenitor cells (hAPCs). Unfortunately, these display replicative senescence after prolonged culture conditions, which limits their use in mechanistic studies. METHODS: Herein, we knocked in 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 differentiation into adipocytes. RESULTS: Immortalized TERT-hAPCs retained proliferative and adipogenic capacities comparable to those of early-passage wild type hAPCs for > 80 passages. In line with this, our integrative transcriptomic and proteomic analyses revealed that TERT-hAPCs displayed robust adipocyte expression profiles in comparison to wild type hAPCs. This was confirmed by functional analyses of lipid turnover where TERT-hAPCs exhibited pronounced responses to insulin and pro-lipolytic stimuli such as isoprenaline, dibutyrul cAMP and tumour necrosis factor alpha. In addition, TERT-hAPCs could be readily cultured in both standard 2D and 3D-cultures and proteomic analyses revealed that the spheroid culture conditions improved adipogenesis. CONCLUSION: Through descriptive and functional studies, we demonstrate that immortalization of human CD55+ hAPCs is feasible and results in cells with stable proliferative and adipogenic capacities over multiple passages. As these cells are cryopreservable, they provide the additional advantage over primary cells of allowing repeated studies in both 2D and 3D model systems with the same genetic background. (234/250).
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Publication type
Article: Journal article
Document type
Scientific Article
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Keywords
Adipocyte ; Adipogenesis ; Cell Model ; Crispr/cas9 ; Fat Cell ; Genetic Engineering ; Lipolysis ; Multiomics ; Spheroid; Stem-cells; Differentiation
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Language
english
Publication Year
2023
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0
HGF-reported in Year
2023
ISSN (print) / ISBN
2162-3945
e-ISSN
2162-397X
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Volume: 14,
Issue: 1,
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Article Number: 2283213
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Landes Bioscience
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530 Walnut Street, Ste 850, Philadelphia, Pa 19106 Usa
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Peer reviewed
POF-Topic(s)
30201 - Metabolic Health
90000 - German Center for Diabetes Research
Research field(s)
Helmholtz Diabetes Center
PSP Element(s)
G-502200-001
G-501900-221
Grants
Swedish Research Council
Stockholm Region
Novo Nordisk Fonden
Knut och Alice Wallenbergs Stiftelse
European Research Council
European Foundation for the Study of Diabetes (EASD)
Swedish Diabetes Foundation
Center for Innovative Medicine (CIMED)
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Erfassungsdatum
2023-12-19