Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct

Robert Bender; Michelle McCarthy; Theodore Brown; Joanna Bukowska; Stanley Smith; Rosalyn D Abbott; David L Kaplan; Christopher Williams; James W Wade; Andrea Alarcon; Xiying Wu; Frank Lau; Jeffrey M Gimble; Trivia Frazier

doi:10.1155/2020/4242130

Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct

Stem Cells Int. 2020 Jun 10:2020:4242130. doi: 10.1155/2020/4242130. eCollection 2020.

Authors

Robert Bender^{1

2}, Michelle McCarthy², Theodore Brown², Joanna Bukowska^{2

3}, Stanley Smith¹, Rosalyn D Abbott⁴, David L Kaplan⁵, Christopher Williams⁶, James W Wade⁷, Andrea Alarcon¹, Xiying Wu^{1

8}, Frank Lau⁹, Jeffrey M Gimble^{1

2

8

10

11

12}, Trivia Frazier^{1

8

10}

Affiliations

¹ LaCell LLC, New Orleans LA, USA.
² Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans LA, USA.
³ Polish Academy of Science, Olsztyn, Poland.
⁴ Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh PA, USA.
⁵ Departments of Biomedical Engineering and Chemical and Biological Engineering, Bioengineering and Biotechnology Center, Tufts University, Medford MA, USA.
⁶ Department of Pharmacology, Xavier University of Louisiana, New Orleans LA, USA.
⁷ Plastic and Reconstructive Surgery, Baton Rouge LA, USA.
⁸ Obatala Sciences Inc., New Orleans LA, USA.
⁹ LSU Department of Plastic Surgery, New Orleans, LA, USA.
¹⁰ Department of Structural and Cell Biology, Tulane University School of Medicine, New Orleans LA, USA.
¹¹ Department of Medicine, Tulane University School of Medicine, New Orleans LA, USA.
¹² Department of Surgery, Tulane University School of Medicine, New Orleans LA, USA.

Abstract

Obesity, defined as a body mass index of 30 kg/m² or above, has increased considerably in incidence and frequency within the United States and globally. Associated comorbidities including cardiovascular disease, type 2 diabetes mellitus, metabolic syndrome, and nonalcoholic fatty liver disease have led to a focus on the mechanisms promoting the prevention and treatment of obesity. Commonly utilized in vitro models employ human or mouse preadipocyte cell lines in a 2-dimensional (2D) format. Due to the structural, biochemical, and biological limitations of these models, increased attention has been placed on "organ on a chip" technologies for a 3-dimensional (3D) culture. Herein, we describe a method employing cryopreserved primary human stromal vascular fraction (SVF) cells and a human blood product-derived biological scaffold to create a 3D adipose depot in vitro. The "fat-on-chip" 3D cultures have been validated relative to 2D cultures based on proliferation, flow cytometry, adipogenic differentiation, confocal microscopy/immunofluorescence, and functional assays (adipokine secretion, glucose uptake, and lipolysis). Thus, the in vitro culture system demonstrates the critical characteristics required for a humanized 3D white adipose tissue (WAT) model.