Shear Force Processing of Lipoaspirates for Stem Cell Enrichment Does Not Affect Secretome of Human Cells Detected by Mass Spectrometry In Vitro

Abstract

Background: Lipofilling is one of the most often performed surgical procedures in plastic and reconstructive surgery. Lipoaspirates provide a ready source of stem cells and secreted factors that contribute to neoangiogenesis and fat graft survival. However, the regulations about the enrichment of these beneficial cells and factors are ambiguous. In this study, the authors tested whether a combination of centrifugation and homogenization allowed the enrichment of viable stem cells in lipoaspirates through the selective removal of tumescent solution, blood, and released lipids without significantly affecting the cell secretome. Methods: Human lipoaspirate was harvested from six different patients using water jet–assisted liposuction. Lipoaspirate was homogenized by first centrifugation (3584 rpm for 2 minutes), shear strain (10 times intersyringe processing), and second centrifugation (3584 rpm for 2 minutes). Stem cell enrichment was shown by cell counting after stem cell isolation. Lipoaspirate from different processing steps (unprocessed, after first centrifugation, after homogenization, after second centrifugation) was incubated in serum-free cell culture medium for mass spectrometric analysis of secreted proteins. Results: Lipoaspirate homogenization leads to a significant 2.6 ± 1.75-fold enrichment attributable to volume reduction without reducing the viability of the stem cells. Protein composition of the secretome did not change significantly after tissue homogenization. Considering the enrichment effects, there were no significant differences in the protein concentration of the 83 proteins found in all processing steps. Conclusions: Stem cells can be enriched mechanically without significantly affecting the composition of secreted proteins. Shear-assisted enrichment of lipoaspirate constitutes no substantial manipulation of the cells’ secretome.

Systematic review: Advances of fat tissue engineering as bioactive scaffold, bioactive material, and source for adipose-derived mesenchymal stem cells in wound and scar treatment

“…The significance of this observation, however, may be undermined by a relatively small study population. In vitro studies demonstrated that similar modifications increase AD-MSC output [33,[38][39][40][41] without affecting the composition of secreted proteins [41].…”

Facial Rejuvenation with Concentrated Lipograft—A 12 Month Follow-Up Study

“…There have also been apprehensions that mechanical processing might affect the genome and therefore protein synthesis in adipose cells. An analysis of cells in lipoaspirate after mechanical processing showed that mechanical processing does not affect the secretome of autologous adipose cells and can therefore be used safely in a multivariety of surgical settings [ 37 ]. On top, the authors managed to show that mechanical processing was able to boost the cellular fraction in comparison to nonprocessed lipoaspirate.…”

Adipose Tissue-Derived Stem Cell Yield Depends on Isolation Protocol and Cell Counting Method

“…Evidently, ASCs contribute to fat graft survival not only by their proliferative capacities but also by paracrine angiogenic properties [ 9 , 41 ]. Therefore, enrichment of lipoaspirate with ASCs is intensively investigated by numerous groups [ 36 , 42 , 43 , 44 , 45 , 46 ]. Safety concerns regarding enzymatic digestion of fat tissue have led to an increasing recognition of methods that circumvent the risks of enzymatic digestion [ 29 ].…”