Abstract
Adipose tissue is an abundant source of stem cells. However, liposuction cannot yield cell quantities sufficient for direct applications in regenerative medicine. Therefore, the development of GMP-compliant ex vivo expansion protocols is required to ensure the production of a “cell drug” that is safe, reproducible, and cost-effective. Thus, we developed our own basal defined xeno- and serum-free cell culture medium (UrSuppe), specifically formulated to grow human adipose stem cells (hASCs). With this medium, we can directly culture the stromal vascular fraction (SVF) cells in defined cell culture conditions to obtain hASCs. Cells proliferate while remaining undifferentiated, as shown by Flow Cytometry (FACS), Quantitative Reverse Transcription PCR (RT-qPCR) assays, and their secretion products. Using the UrSuppe cell culture medium, maximum cell densities between 0.51 and 0.80 × 105 cells/cm2 (=2.55–4.00 × 105 cells/mL) were obtained. As the expansion of hASCs represents only the first step in a cell therapeutic protocol or further basic research studies, we formulated two chemically defined media to differentiate the expanded hASCs in white or beige/brown adipocytes. These new media could help translate research projects into the clinical application of hASCs and study ex vivo the biology in healthy and dysfunctional states of adipocytes and their precursors. Following the cell culture system developers’ practice and obvious reasons related to the formulas’ patentability, the defined media’s composition will not be disclosed in this study.
Highlights
Adipose tissue is usually classified by morphology into white, brown, or beige subsets [1,2]
As a first step in this direction, we developed xeno- and serum-free media, which induce the maturation of human adipose stem cells (hASCs) into white or beige adipocytes and which we named “UrSuppe-white adipose tissue” (US-WAT) and “UrSuppe-beige/brown adipose tissue”
As a first very positive result, we found that two signature adipokines for differentiated cells were very low or missing in the supernatant of non-induced hASCs, while present in cells cultured with the WAT or brown adipose tissue (BAT)
Summary
Adipose tissue is usually classified by morphology into white, brown, or beige subsets [1,2]. The primary white adipose tissue (WAT) depot is further subdivided as either subcutaneous (SAT) or visceral (VAT). The minor white adipose depots include the dermal WAT (dWAT) and bone marrow adipose tissue (MAT), which are considered distinct from SAT and VAT [3]. Rodents are commonly used to study human metabolism and obesity. It is unclear to what extent rodent fat pads are a suitable human adipogenesis model in health and disease. There are essential anatomical differences that should be considered [4,5].
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