Abstract
Stem cells derived from the subcutaneous adipose tissue of debrided burned skin represent an appealing source of adipose-derived stem cells (ASCs) for regenerative medicine. Traditional tissue culture uses fetal bovine serum (FBS), which complicates utilization of ASCs in human medicine. Human platelet lysate (hPL) is one potential xeno-free, alternative supplement for use in ASC culture. In this study, adipogenic and osteogenic differentiation in media supplemented with 10% FBS or 10% hPL was compared in human ASCs derived from abdominoplasty (HAP) or from adipose associated with debrided burned skin (BH). Most (95–99%) cells cultured in FBS were stained positive for CD73, CD90, CD105, and CD142. FBS supplementation was associated with increased triglyceride content and expression of adipogenic genes. Culture in hPL significantly decreased surface staining of CD105 by 31% and 48% and CD142 by 27% and 35% in HAP and BH, respectively (p < 0.05). Culture of BH-ASCs in hPL also increased expression of markers of osteogenesis and increased ALP activity. These data indicate that application of ASCs for wound healing may be influenced by ASC source as well as culture conditions used to expand them. As such, these factors must be taken into consideration before ASCs are used for regenerative purposes.
Highlights
Adipose tissue is a rich reservoir of mesenchymal stem cells (ASCs) with a high self-renewing capacity [1,2,3,4]
We investigated the functionality of ASCs derived from discarded human burn tissue (BH) from three individual patients (n = 3) ranging in age from 25 to 75 years of age and ASCs from subcutaneous adipose tissue from three abdominoplasty patients (HAP; n = 3) of approximately 45–60 years of age
Our study shows the following: (i) while fetal bovine serum (FBS) supports adipogenic differentiation in both groups, Human platelet lysate (hPL) may enhance basal lipid accumulation while hindering terminal adipogenesis, (ii) there is a higher inherent level of osteogenic transcripts in ASCs from burn samples, which is highly sensitive to external stimuli and to the factors within hPL, (iii) ASCs derived from abdominoplasty are less sensitive to osteogenic stimulus in comparison to Burned human skin ASCs (BH) ASCs but are not significantly different from the commercial ASCs from a healthy individual, and (iv) there are differences among ASCs that are dependent upon the donor, as evidenced by individual proliferation rates and CD142 surface staining (Supplemental Figure 2 and Supplemental Figure 3) and source that may influence differentiation potential
Summary
Adipose tissue is a rich reservoir of mesenchymal stem cells (ASCs) with a high self-renewing capacity [1,2,3,4]. Due to the unique ability of ASCs to elicit this beneficial healing response, clinical studies have been conducted using autologous stem cells for regenerative therapies [14,15,16,17,18]. According to the International Society of Cellular Therapy (ISCT) and International Federation of Adipose Therapeutics and Sciences (IFATS) panel of experts, freshly isolated stromal vascular fraction, as well as culture-expanded stem cells, are expected to express a positive panel of surface markers—CD13, CD73, CD90, and CD105—and remain negative for markers like CD31 and CD45. The stem cells must be checked for their ability to functionally differentiate into osteogenic, adipogenic, and chondrogenic lineages [19]
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