Abstract
BackgroundAdipose tissue contains a large number of multipotent cells, which are essential for stem cell-based therapies. The combination of this therapy with suitable commercial clinically used matrices, such as collagen and elastin matrices (i.e. dermal matrices), is a promising approach for soft tissue reconstruction. We previously demonstrated that the liposuction method affects the adherence behaviour of freshly isolated adipose-derived stem/stromal cells (ASCs) on collagen and elastin matrices. However, it remains unclear whether freshly isolated and uncultured ASCs could be directly transferred to matrices during a single transplantation operation without additional cell culture steps.MethodsAfter each fat harvesting procedure, ASCs were isolated and directly seeded onto collagen and elastin matrices. Different time intervals (i.e. 1, 3 and 24 h) were investigated to determine the time interval needed for cellular attachment to the collagen and elastin matrices. Resazurin-based vitality assays were performed after seeding the cells onto the collagen and elastin matrices. In addition, the adhesion and migration of ASCs on the collagen and elastin matrices were visualised using histology and two-photon microscopy.ResultsA time-dependent increase in the number of viable ASCs attached to the collagen and elastin matrices was observed. This finding was supported by mitochondrial activity and histology results. Importantly, the ASCs attached and adhered to the collagen and elastin matrices after only 1 h of ex vivo enrichment. This finding was also supported by two-photon microscopy, which revealed the presence and attachment of viable cells on the upper layer of the construct.ConclusionFreshly isolated uncultured ASCs can be safely seeded onto collagen and elastin matrices for ex vivo cellular enrichment of these constructs after liposuction. Although we observed a significant number of seeded cells on the matrices after a 3-h enrichment time, we also observed an adequate number of isolated cells after a 1-h enrichment time. However, this approach must be optimised for clinical use. Thus, in vivo studies and clinical trials are needed to investigate the feasibility of this approach.
Highlights
Adipose tissue contains a large number of multipotent cells, which are essential for stem cell-based therapies
This fraction can be obtained through an isolation process that uses collagenase secondary to liposuction, resulting in a component that contains multiple cells: adipose-derived stem/stromal cells (ASCs)
The green colour indicates the cytoplasmic portion of cells, which was stained with fluorescein diacetate (FDA), and the blue colour shows the nuclei, which were stained with Hoechst 33342
Summary
Adipose tissue contains a large number of multipotent cells, which are essential for stem cell-based therapies. We previously demonstrated that the liposuction method affects the adherence behaviour of freshly isolated adipose-derived stem/stromal cells (ASCs) on collagen and elastin matrices. It remains unclear whether freshly isolated and uncultured ASCs could be directly transferred to matrices during a single transplantation operation without additional cell culture steps. The term SVF is commonly used in the literature and refers to the cellular pellet without fat cells (i.e., mature adipocytes) This fraction can be obtained through an isolation process that uses collagenase secondary to liposuction (i.e., fat harvesting), resulting in a component that contains multiple cells: adipose-derived stem/stromal cells (ASCs). Adipose-derived stem cells are adult self-renewing cells of mesenchymal origin that can differentiate into the adipogenic lineage and into the osteogenic, chondrogenic, myogenic and neurogenic lineages in vitro [3,4]
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