Abstract
Human adipose-derived stem cells (ASCs) have potential to improve wound healing; however, their equivalents from domestic animals have received less attention as an alternative cell-based therapy for animals or even humans. Hypoxia is essential for maintaining stem cell functionality in tissue-specific niches. However, a cellular response to low oxygen levels has not been demonstrated in pig ASCs. Hence, the goal of our study was to characterize ASCs isolated from the subcutaneous fat of domestic pigs (pASCs) and examine the effect of hypoxia on their proteome and functional characteristics that might reproduce pASCs wound healing ability. Analysis of immunophenotypic and functional markers demonstrated that pASCs exhibited characteristics of mesenchymal stem cells. Proteomic analysis revealed 70 differentially abundant proteins between pASCs cultured under hypoxia (1% O2) or normoxia (21% O2). Among them, 42 proteins were enriched in the cells exposed to low oxygen, whereas 28 proteins showed decrease expression following hypoxia. Differentially expressed proteins were predominantly involved in cell metabolism, regulation of focal and intracellular communication, and attributed to wound healing. Functional examination of hypoxic pASCs demonstrated acquisition of contractile abilities in vitro. Overall, our results demonstrate that hypoxia pre-conditioning impacts the pASC proteome signature and contractile function in vitro and hence, they might be considered for further cell-based therapy study on wound healing.
Highlights
Adipose-derived stem cells (ASCs) are increasingly recognized as a vital cell source for cell-based therapy in regenerative medicine[1,2,3]
Clonogenicity assays revealed that pASCs plated at low concentrations (10, 100, or 200 cells/well) demonstrated the potential to form colony-forming units (CFUs) (Fig. 1D), while high-density plating (400 or 1000 cells/well) led to reduced clonogenicity regarding the extent of establishing of a cell monolayer (Fig. 1D). pASCs exposed to adipogenic or osteogenic differentiationinduction media displayed the capacity to differentiate into adipocytes or osteoblasts as evidenced by histological examination of lipid droplets stained with Oil Red O and calcium deposits stained with Alizarin Red S, respectively (Fig. 1E)
In searching for alternative sources of stem cells to facilitate wound healing in animals and/or humans, we isolated and characterized of adipose-derived stem cells (ASCs) from the subcutaneous fat of domestic pigs and showed that they fulfilled the criteria of MSCs
Summary
Adipose-derived stem cells (ASCs) are increasingly recognized as a vital cell source for cell-based therapy in regenerative medicine[1,2,3]. Frazier et al.[19] showed that pre-conditioning of human ASCs with low oxygen reduced secretion of extracellular matrix (ECM) proteins and type 2 cytokines (interleukin [IL]-6, IL-13, monocyte chemoattractant protein [MCP]-1, CD40 ligand) involved in the initiation of fibrosis. These results suggest that enhancing these ASCs characteristics with hypoxia may increase their regenerative potential and augment healing following cell delivery to the injured tissues. The hypothesis that hypoxic pre-conditioning of ASCs provides functional e nhancement[15,18,20] and makes the cells more attractive for clinical translational science has encouraged an interest in characterizing the hypoxiaresponsive characteristics of ASCs on the proteomic level[21]. There are no data related to effect of hypoxia on pig ASCs proteome and the cell function
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