Immunophenotyping of a Stromal Vascular Fraction from Microfragmented Lipoaspirate Used in Osteoarthritis Cartilage Treatment and Its Lipoaspirate Counterpart.

Polancec Polancec,Primorac Primorac,Boric Boric,Rod Rod,Nogalo Nogalo,Skelin Skelin,Jelec Jelec,Zenic Zenic,Plecko Plecko,Turkalj Turkalj,Hudetz Hudetz,Vrdoljak Vrdoljak

doi:10.3390/genes10060474

Abstract

Osteoarthritis (OA) is a degenerative joint disease accompanied by pain and loss of function. Adipose tissue harbors mesenchymal stem/stromal cells (MSC), or medicinal signaling cells as suggested by Caplan (Caplan, 2017), used in autologous transplantation in many clinical settings. The aim of the study was to characterize a stromal vascular fraction from microfragmented lipoaspirate (SVF-MLA) applied for cartilage treatment in OA and compare it to that of autologous lipoaspirate (SVF-LA). Samples were first stained using a DuraClone SC prototype tube for the surface detection of CD31, CD34, CD45, CD73, CD90, CD105, CD146 and LIVE/DEAD Yellow Fixable Stain for dead cell detection, followed by DRAQ7 cell nuclear dye staining, and analyzed by flow cytometry. In SVF-LA and SVF-MLA samples, the following population phenotypes were identified within the CD45− fraction: CD31+CD34+CD73±CD90±CD105±CD146± endothelial progenitors (EP), CD31+CD34−CD73±CD90±CD105−CD146± mature endothelial cells, CD31−CD34−CD73±CD90+CD105−CD146+ pericytes, CD31−CD34+CD73±CD90+CD105−CD146+ transitional pericytes, and CD31−CD34+CD73highCD90+CD105−CD146− supra-adventitial-adipose stromal cells (SA-ASC). The immunophenotyping profile of SVF-MLA was dominated by a reduction of leukocytes and SA-ASC, and an increase in EP, evidencing a marked enrichment of this cell population in the course of adipose tissue microfragmentation. The role of EP in pericyte-primed MSC-mediated tissue healing, as well as the observed hormonal implication, is yet to be investigated.

Highlights

Ever since the discovery that human adipose tissue is a rich source of mesenchymal stem/stromal cells (MSC) and is bestowed with therapeutic potential after culture expansion and differentiation [1,2,3], autologous adipose tissue has been explored for its regenerative properties in many clinical settings
CD45+ stromal vascular fraction (SVF) cells were determined in both lipoaspirate (SVF-LA) and its microfragmented lipoaspirate counterpart (SVF-MLA), and viability determined
In an attempt to contribute to the growing need to understand the adipose tissue MSC used in therapeutic intervention, we investigated the cell content of the MLA product applied for the therapeutic intervention, we investigated the cell content of the MLA product applied for the treatment treatment of OA patients and its lipoaspirate counterpart [9,15]

Summary

Introduction

Ever since the discovery that human adipose tissue is a rich source of mesenchymal stem/stromal cells (MSC) and is bestowed with therapeutic potential after culture expansion and differentiation [1,2,3], autologous adipose tissue has been explored for its regenerative properties in many clinical settings.MSC are contained within the stromal vascular fraction (SVF)—the aqueous portion obtained after enzymatic digestion of lipoaspirate (LA)—together with other cells of the differentiated or multipotent features. Ever since the discovery that human adipose tissue is a rich source of mesenchymal stem/stromal cells (MSC) and is bestowed with therapeutic potential after culture expansion and differentiation [1,2,3], autologous adipose tissue has been explored for its regenerative properties in many clinical settings. Lipogems® technology has emerged as a platform that yields an intact SVF and MSC of a high therapeutic potential (reviewed in [6]), obtained by the microfragmentation of autologous fat tissue [7]. The use of such microfragmented lipoaspirate (MLA) has reached a fruitful application in osteoarthritis (OA), one of the leading musculoskeletal disorders in the adult population worldwide [8].

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