Characterization of 3D Organotypic Culture of Mouse Adipose-Derived Stem Cells

• Published in: International journal of molecular sciences Vol. 25; no. 7; p. 3931
• Main Authors: Son, Tae Gen, Seo, Yoojin, Kim, Won-Tae, Kim, Meesun, Choi, Seon Jeong, Choi, Si Ho, Sung, Byung-Jun, Min, Jae-Seok, Han, Eon Chul, Kim, Hyung-Sik
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.04.2024
• Subjects: Adipocytes; adipogenic differentiation; Adipose tissues; Adiposity; Animals; Body fat; Cell culture; Cell cycle; Gene expression; Genes; Investigations; Lipoprotein lipase; mesenchymal–epithelial transition (MET); Metabolism; Mice; Microphysiological Systems; Obesity; Organoids; Regenerative medicine; regenerative potential; Scientific equipment and supplies industry; Senescence; Stem cells; three-dimensional (3D) culture; United States; South Korea; regenerative potential; three-dimensional (3D) culture; mesenchymal–epithelial transition (MET); adipogenic differentiation
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25073931

Although stem cells are a promising avenue for harnessing the potential of adipose tissue, conventional two-dimensional (2D) culture methods have limitations. This study explored the use of three-dimensional (3D) cultures to preserve the regenerative potential of adipose-derived stem cells (ADSCs) and investigated their cellular properties. Flow cytometric analysis revealed significant variations in surface marker expressions between the two culture conditions. While 2D cultures showed robust surface marker expressions, 3D cultures exhibited reduced levels of CD44, CD90.2, and CD105. Adipogenic differentiation in 3D organotypic ADSCs faced challenges, with decreased organoid size and limited activation of adipogenesis-related genes. Key adipocyte markers, such as lipoprotein lipase (LPL) and adipoQ, were undetectable in 3D-cultured ADSCs, unlike positive controls in 2D-cultured mesenchymal stem cells (MSCs). Surprisingly, 3D-cultured ADSCs underwent mesenchymal-epithelial transition (MET), evidenced by increased E-cadherin and EpCAM expression and decreased mesenchymal markers. This study highlights successful ADSC organoid formation, notable MSC phenotype changes in 3D culture, adipogenic differentiation challenges, and a distinctive shift toward an epithelial-like state. These findings offer insights into the potential applications of 3D-cultured ADSCs in regenerative medicine, emphasizing the need for further exploration of underlying molecular mechanisms.