Abstract
This work aims to characterize a new method to recover low-manipulated human adipose tissue, enriched with adipose tissue-derived mesenchymal stem cells (ATD-MSCs) for autologous use in regenerative medicine applications. Lipoaspirated fat collected from patients was processed through Lipocell, a Class II-a medical device for dialysis of adipose tissue, by varying filter sizes and washing solutions. ATD-MSC yield was measured with flow cytometry after stromal vascular fraction (SVF) isolation in fresh and cultured samples. Purification from oil and blood was measured after centrifugation with spectrophotometer analysis. Extracellular matrix preservation was assessed through hematoxylin and eosin (H&E) staining and biochemical assay for total collagen, type-2 collagen, and glycosaminoglycans (GAGs) quantification. Flow cytometry showed a two-fold increase of ATD-MSC yield in treated samples in comparison with untreated lipoaspirate; no differences where reported when varying filter size. The association of dialysis and washing thoroughly removed blood and oil from samples. Tissue architecture and extracellular matrix integrity were unaltered after Lipocell processing. Dialysis procedure associated with Ringer’s lactate preserves the proliferation ability of ATD-MSCs in cell culture. The characterization of the product showed that Lipocell is an efficient method for purifying the tissue from undesired byproducts and preserving ATD-MSC vitality and extracellular matrix (ECM) integrity, resulting in a promising tool for regenerative medicine applications.
Highlights
Adipose tissue and its derivatives are being increasingly used as autografts
Cellular phenotyping was performed after enzymatic stromal vascular fraction (SVF) isolation on both untreated lipoaspirated adipose tissue and Lipocell-processed products
The combination of dialysis and brushing of the Lipocell procedure retains the cellular contents of the adipose tissue without significant loss; the quantification of viable cells normalized for the adipose tissue weight showed no differences (Figure 1a) among the three filters (15 μm, 20 μm, and 50 μm)
Summary
Adipose tissue and its derivatives are being increasingly used as autografts. Bioactive fractions or transformed products of fat include microfat, nanofat, stromal vascular fraction (SVF), and adipose-derived stem cells (ATD-MSC). Indications for the use of these autologous tissue derivatives are several and enlarging [1], i.e., cosmetic surgery for creating, increasing, or reconstructing volumes, Processes 2020, 8, 88; doi:10.3390/pr8010088 www.mdpi.com/journal/processes. As with any other tissue for immediate autografting, users must respect minimal manipulation requirements. The enzymatic digestion of adipose tissue, which disrupts the extracellular matrix (ECM) with the release of SVF, is considered high manipulation and must be performed into cell factories. The FDA considers that significant physical processing is more than minimal manipulation when the structural integrity of the tissue is altered
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