Adipose Tissue-Derived Stem Cells Retain Their Adipocyte Differentiation Potential in Three-Dimensional Hydrogels and Bioreactors

Abstract

Osteoarthritis (OA) is a common joint disorder with a significant economic and healthcare impact. The knee joint is composed of cartilage and the adjoining bone, a synovial capsule, the infrapatellar fat pad (IPFP), and other connective tissues such as tendons and ligaments. Adipose tissue has recently been highlighted as a major contributor to OA through strong inflammation mediating effects. In this study, methacrylated gelatin (GelMA) constructs seeded with adipose tissue-derived mesenchymal stem cells (ASCs) and cultured in a 3D printed bioreactor were investigated for use in microphysiological systems to model adipose tissue in the knee joint. Four patient-derived ASC populations were seeded at a density of 20 million cells/mL in GelMA. Live/Dead and boron-dipyrromethene/4′,6-diamidino-2-phenylindole (BODIPY/DAPI) staining of cells within the constructs demonstrated robust cell viability after 28 days in a growth (control) medium, and robust cell viability and lipid accumulation in adipogenic differentiation medium. qPCR gene expression analysis and protein analysis demonstrated an upregulated expression of key adipogenesis-associated genes. Overall, these data indicate that ASCs retain their adipogenic potential when seeded within GelMA hydrogels and cultured within perfusion bioreactors, and thus can be used in a 3D organ-on-a-chip system to study the role of the IPFP in the pathobiology of the knee OA.

Caveolin-1 mediates soft scaffold-enhanced adipogenesis of human mesenchymal stem cells

“…Adipose tissue engineering provides a robust tool to regenerate lost or damaged fat. In vitro created adipose tissue has recently been used to model diseases and develop drugs [ 27 , 30 – 32 ]. There is thus a demonstrated need for functional fat tissues generated through tissue engineering.…”

“…Its recognized biosafety makes it a convenient biomaterial to generate adipose tissues for different applications. To date, gelatin sponge or gelatin hydrogels have been used to create adipose tissues from human MSCs [ 32 , 34 , 35 ]. In these studies, MSCs displayed reduced adipogenic potential in gelatin cultures when compared to monolayer cultures.…”

Human Adipose-Derived Stromal/Stem Cell Culture and Analysis Methods for Adipose Tissue Modeling In Vitro: A Systematic Review

“…Finally, none of the screened articles exclusively studied 2D culture; therefore, all articles included in this analysis report 3D culture methods. Data and analysis hereafter exclusively refer to the 27 articles [4,5,[8][9][10][11][12][13][15][16][17][19][20][21][22][23][24][25][26]28,30,31,33,38,44,45,48] identified to be most relevant to the central goal of this systematic review. Out of the 27 relevant articles, 26 articles used adipogenic medium to differentiate hASC into adipocytes in vitro.…”

“…Flow cytometry is a widely used assay for assessing the immunophenotype of mesenchymal stromal/stem cells (MSCs). According to the International Federation for Adipose Therapeutics (IFATS) and Science and the International Society for Cellular Therapy (ISCT), hASC in culture maintain markers similar to other MSCs as they are positive for CD90, CD73, CD105, and CD44 and negative for CD45 and CD31 [25]. Flow cytometry It can be challenging to image adipocytes using light microscopy because of their high lipid content which causes autofluorescence that is enhanced by light diffraction.…”

Breast Cancer Reconstruction: Design Criteria for a Humanized Microphysiological System

“…For example, the successful construction of both an adipose and bone marrow MPS have been described recently in the literature. [50][51][52][53] Using such MPS constructs, it would be possible to seed an adipose MPS with a primary breast cancer tumor and to monitor its ability to metastasize through a microperfusion circulatory system into a parallel bone marrow MPS. Alternatively, tissue from a bone marrow metastatic breast tumor could be seeded directly into a bone marrow microenvironment MPS.…”