Abstract
Hydrogels serve as three-dimensional scaffolds whose composition can be customized to allow attachment and proliferation of several different cell types. Extracellular matrix-derived hydrogels are considered close replicates of the tissue microenvironment. They can serve as scaffolds for in vitro tissue engineering and are a useful tool to study cell-scaffold interaction. The aim of the present study was to analyze the effect of adipose-derived stromal/stem cells (ASCs) and decellularized adipose tissue-derived (DAT) hydrogel interaction on ASC morphology, proliferation, differentiation, and DAT hydrogel microstructure. First, the ASCs were characterized using flow cytometry, adipogenic/osteogenic differentiation, colony-forming unit fibroblast assay and doubling time. The viability and proliferation assays showed that ASCs seeded in DAT hydrogel at different concentrations and cultured for 21 days remained viable and displayed proliferation. ASCs were seeded on DAT hydrogel and cultured in stromal, adipogenic, or osteogenic media for 14 or 28 days. The analysis of adipogenic differentiation demonstrated the upregulation of adipogenic marker genes and accumulation of oil droplets in the cells. Osteogenic differentiation demonstrated the upregulation of osteogenic marker genes and mineral deposition in the DAT hydrogel. The analysis of DAT hydrogel fiber metrics revealed that ASC seeding, and differentiation altered both the diameter and arrangement of fibers in the matrix. Matrix metalloproteinase-2 (MMP-2) activity was assessed to determine the possible mechanism for DAT hydrogel remodeling. MMP-2 activity was observed in all ASC seeded samples, with the osteogenic samples displaying the highest MMP-2 activity. These findings indicate that DAT hydrogel is a cytocompatible scaffold that supports the adipogenic and osteogenic differentiation of ASCs. Furthermore, the attachment of ASCs and differentiation along adipogenic and osteogenic lineages remodels the microstructure of DAT hydrogel.
Highlights
Human adipose-derived stromal/stem cells (ASCs) are extracted from adipose tissue by collagenase digestion [1]
Based on flow cytometry analysis, the ASC population was found to be positive for phenotypic markers CD73, CD90, and CD105 (Figure 1(a)), while negative for CD3, CD14, CD31, and CD45 (Figure 1(b)), consistent with the immunophenotype defined for ASC
Adipogenic differentiation of ASCs after 28 days of induction was confirmed by Oil Red O staining, where the cells stained positive for intracellular lipid vacuoles (Figure 1(c))
Summary
Human adipose-derived stromal/stem cells (ASCs) are extracted from adipose tissue by collagenase digestion [1]. In an effort to recapitulate in vivo conditions, recent research efforts have focused on culturing ASCs in three-dimensional (3D) systems like spheroids and hydrogels [5,6,7]. A hydrogel of decellularized adipose tissue (DAT) can serve as an alternative ECM for research, due to the abundant availability of source tissue [11, 12] and accumulating evidence of its cytocompatibility [13]. DAT hydrogel and its composites support in vitro ASC proliferation [14] and adipogenic differentiation [15,16,17] and have proven to be adipoinductive scaffolds [9, 18,19,20]. Intact DAT scaffolds have displayed positive outcomes in wound healing [21] and nerve repair [22]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
