Comparison of Growth Supplements on the Differentiation of Adipose Derived Stem Cells from Burned and Non‐Burned Human Skin

Severe traumatic injuries, such as large total body surface area burns, limit the availability of autologous cells for wound repair. Previous experiments from our lab have demonstrated that adipose derived stem cells (ASCs) from burned debrided skin are viable, display an immunophenotype indicative of stemness, and are capable of tri‐lineage differentiation. Traditional tissue culture utilizes undefined, animal‐derived components that present numerous complications for the utilization of ASCs in human medicine. Accordingly, there is a need for defined, xeno‐free cell culture media. Human platelet lysate (hPL) is one potential media supplement for the replacement of fetal bovine serum in stem cell culture. In this study, burned human ASCs (BH‐ASCs) and ASCs derived from abdominoplasty (HAP‐ASCs) were compared in regard to the ability to proliferate and differentiate in media supplemented with traditional 10% FBS, 10% hPL, or grown in MesenPRO RS™. ASCs were obtained from the stromal vascular fractions of subcutaneous adipose tissue from burned and non‐burned skin. Cells were cultured in DMEM with 10% FBS or 10% hPL, or in MesenPRO RS™. Cells were induced to differentiate towards an adipogenic or osteogenic lineage at an early passage utilizing traditional differentiation media supplemented with FBS or 10% hPL. DMEM or αMEM was used for adipogenic and osteogenic lineages, respectively, with the addition of standard induction agents. Adipogenic differentiation was assessed after 14 days by Oil red O staining, triglyceride content, and expression of markers of adipogenesis, including PPARγ, leptin, and adiponectin. Osteogenic differentiation was assessed by alizarin red S staining and expression of markers of ostogenesis, including BMP‐2 and RUNX2. Results indicate that adipogenic and osteogenic differentiation appears to be influenced by both the ASC source (burned vs. non‐burned skin) and by the supplement in which they are grown and differentiated. Use of MesenPRO RS™ and supplementation with hPL appears to enhance differentiation towards both lineages over standard FBS. Prior experiments in our labs have indicated that both burn and non‐burn ACSs are capable of trilineage differentiation, however there is a need to characterize these cells, particularly the BH‐ASCs in xeno‐free media, with regard to their stemness and immunosuppresive capabilites if they are to be utilized in any capacity for regenerative medicine. Ongoing research efforts in our laboratory seek to address these issues to better define the utility of BH‐ASCs as a burn wound therapeutic. Support or Funding Information This work was possible due to the generous funding from the US Army Medical Research and Material Command and ORISE