An organoid model derived from human adipose stem/progenitor cells to study adipose tissue physiology

Autor: Markus Mandl, Hans P. Viertler, Florian M. Hatzmann, Camille Brucker, Sonja Großmann, Petra Waldegger, Tina Rauchenwald, Monika Mattesich, Marit Zwierzina, Gerhard Pierer, Werner Zwerschke
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Adipocyte, Vol 11, Iss 1, Pp 164-174 (2022)
Druh dokumentu: article
ISSN: 21623945
2162-397X
2162-3945
DOI: 10.1080/21623945.2022.2044601
Popis: We established a functional adipose organoid model system for human adipose stem/progenitor cells (ASCs) isolated from white adipose tissue (WAT). ASCs were forced to self-aggregate by a hanging-drop technique. Afterwards, spheroids were transferred into agar-coated cell culture dishes to avoid plastic-adherence and dis-aggregation. Adipocyte differentiation was induced by an adipogenic hormone cocktail. Morphometric analysis revealed a significant increase in organoid size in the course of adipogenesis until d 18. Whole mount staining of organoids using specific lipophilic dyes showed large multi- and unilocular fat deposits in differentiated cells indicating highly efficient differentiation of ASCs into mature adipocytes. Moreover, we found a strong induction of the expression of key adipogenesis and adipocyte markers (CCAAT/enhancer-binding protein (C/EBP) β, peroxisome proliferator-activated receptor (PPAR) γ, fatty acid-binding protein 4 (FABP4), adiponectin) during adipose organoid formation. Secreted adiponectin was detected in the cell culture supernatant, underscoring the physiological relevance of mature adipocytes in the organoid model. Moreover, colony formation assays of collagenase-digested organoids revealed the maintenance of a significant fraction of ASCs within newly formed organoids. In conclusion, we provide a reliable and highly efficient WAT organoid model, which enables accurate analysis of cellular and molecular markers of adipogenic differentiation and adipocyte physiology.
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