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

Abstract

Human adipose-derived stromal/stem cells (hASC) are widely used for in vitro modeling of physiologically relevant human adipose tissue. These models are useful for the development of tissue constructs for soft tissue regeneration and 3-dimensional (3D) microphysiological systems (MPS) for drug discovery. In this systematic review, we report on the current state of hASC culture and assessment methods for adipose tissue engineering using 3D MPS. Our search efforts resulted in the identification of 184 independent records, of which 27 were determined to be most relevant to the goals of the present review. Our results demonstrate a lack of consensus on methods for hASC culture and assessment for the production of physiologically relevant in vitro models of human adipose tissue. Few studies have assessed the impact of different 3D culture conditions on hASC adipogenesis. Additionally, there has been a limited use of assays for characterizing the functionality of adipose tissue in vitro. Results from this study suggest the need for more standardized culture methods and further analysis on in vitro tissue functionality. These will be necessary to validate the utility of 3D MPS as an in vitro model to reduce, refine, and replace in vivo experiments in the drug discovery regulatory process.

Adipose Stem Cells in Regenerative Medicine: Looking Forward

“…Scaffolds, synthetic or natural “biologically-derived”, to be used in tissue engineering applications, must recapitulate the extracellular matrix (ECM) and imitate an in vivo like microenvironment favorable for stem cell attachment and proliferation. Synthetic scaffolds are made up of polyesters, polyethers, polyethylene glycol and polylactic acid (PLLA) ( Gibler et al, 2021b ; Reddy et al, 2021 ), while natural scaffolds “commonly used with ASCs” are comprised of collagen, fibrin, gelatin, vitronectin, laminin, alginate, hyaluronic acid, or decellularized materials (DAT) ( Sadeghi-Ataabadi et al, 2017 ; Vinson et al, 2017 ; Kook et al, 2018 ; Mohiuddin O. A. et al, 2019 ; Colle et al, 2020 ; O’Donnell et al, 2020 ).…”

“…In addition, researchers have recently been designing new 3D biomaterials by combining ASCs with biomimetic scaffolds composed of either natural or synthetic materials. These 3D biomaterials have proven effective in tissue repair and organ regeneration ( Storti et al, 2019 ; Gibler et al, 2021 ). These scaffolds have biological and physical properties that imitate the native ECM niche, which is crucial for stem cell adhesion, growth, proliferation and differentiation along particular lineages.…”

Adipose microtissue-on-chip: a 3D cell culture platform for differentiation, stimulation, and proteomic analysis of human adipocytes

“…9 3D cell culture formats, instead of traditional monolayer cultures [10][11][12] have been suggested to enhance the maturation and differentiation of hASCs. 7,13,14 Compared with monolayer culture formats, 3D cell cultures represent the organotypic microenvironment of native adipose tissue. 6 For 3D culture, approaches such as embedding hASCs in Matrigel 15 or other hydrogels, 5,7 low-attachment well plates, 6 hanging drops, 16 magnetic levitation systems, 17 permeable membranes, 18 and microfluidic systems 4,19 have all been exploited.…”

Enhanced Adipogenic Differentiation of Human Dental Pulp Stem Cells in Enzymatically Decellularized Adipose Tissue Solid Foams

“…There is a variety of methodologies to produce in vitro 3D tissue models, which, typically, required the use of biomaterials as scaffolds for cells to grow. Scaffold-based 3D cell cultures are expected to yield results with higher predictive value for clinical outcomes, and are well suited to drug discovery to obtain more accurate results [ 14 , 15 ]. Although substantial advances have been made, the development of an ideal tissue engineered human 3D adipose tissue model is still technically challenging.…”