Polychromatic Flow Cytometric Analysis of Stromal Vascular Fraction from Lipoaspirate and Microfragmented Counterparts Reveals Sex-Related Immunophenotype Differences

Abstract

Mesenchymal stem/stromal cells or medicinal signaling cells (MSC)-based therapy holds promise as a beneficial strategy for treating knee OA (osteoarthritis), but there is no standardized protocols nor mechanistic understanding. In order to gain a better insight into the human MSC from adipose tissue applied for autologous OA treatment, we performed extensive comparative immunophenotyping of the stromal vascular fraction from lipoaspirate or microfragmented lipoaspirates by polychromatic flow cytometry and investigated the cellular components considered responsible for cartilage regeneration. We found an enrichment of the regenerative cellular niche of the clinically applied microfragmented stromal vascular fraction. Sex-related differences were observed in the MSC marker expression and the ratio of the progenitor cells from fresh lipoaspirate, which, in female patients, contained a higher expression of CD90 on the three progenitor cell types including pericytes, a higher expression of CD105 and CD146 on CD31highCD34high endothelial progenitors as well as of CD73 on supra-adventitialadipose stromal cells. Some of these MSC-expression differences were present after microfragmentation and indicated a differential phenotype pattern of the applied MSC mixture in female and male patients. Our results provide a better insight into the heterogeneity of the adipose MSC subpopulations serving as OA therapeutics, with an emphasis on interesting differences between women and men.

Novel Approach in Rectovaginal Fistula Treatment: Combination of Modified Martius Flap and Autologous Micro-Fragmented Adipose Tissue

“…Mesenchymal stem cells (MSCs) are multipotent adult stem cells that have the ability to differentiate into a variety of cell types (adipocytes, osteoblasts, myocytes, chondrocytes, etc.) dictated by the given milieu [ 35 , 40 , 41 , 42 , 47 , 48 , 49 ]. Regarding origin, MSCs can be extracted from the bone marrow (BM-MSCs), umbilical cord MSCs (UC-MSCs), amniotic fluid SCs (AF-SC), placental MSCs (P-MSC), menstrual blood SCs (Men-SC), breast milk, cervix, dental tissue, synovial tissue, and fluid, or can be adipose-derived MSCs (AD-MSCs) and amnion-derived MSCs.…”

“…Via polychromatic flow cytometric analysis, earlier, we determined the phenotypes of the CD45 subpopulation in autologous micro-fragmented samples containing the stromal vascular fraction (SVF). The most predominant cells include 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 (CD31− CD34+ CD73± CD90+ CD105− CD146+), and supra-adventitial-adipose stromal cells (CD31− CD34+ CD73high CD90+ CD105− CD146−) [ 48 , 49 ]. On the other hand, MSCs express very low levels of surface antigens and therefore do not trigger an immune response, allowing their usage as therapeutic agents in regenerative medicine [ 50 , 51 , 52 , 53 ].…”

Evaluation of salivary Thy‐1 in health, periodontitis, and obesity

“…The significant differences between salivary Thy‐1 of females, in the periodontitis group and the other groups, maybe because, generally several innate and adaptive immune responses are greater in females (e.g., macrophage activation, IL‐10 production, T‐, and B‐cell activity, antibody production) when compared with males due to sex‐specific genetic, hormonal and environmental interactions (Klein & Flanagan, 2016). In knee osteoarthritis, women had a higher expression of Thy‐1 from lipoaspirates compared to men (Zenic et al, 2021). Future research can consider sex‐based Thy‐1 variations in periodontitis.…”

Results of Treating Mild to Moderate Knee Osteoarthritis with Autologous Conditioned Adipose Tissue and Leukocyte-Poor Platelet-Rich Plasma

“…While the work by Zenić et al proved that the same population of cells (after treating the samples with 1% collagenase type I) is present in identical ratios in both MFAT and ACA, the results in this study might be explained by the ACA method used in this study as an inefficient methodology in obtaining a significant amount of SVF or by the insufficient cell quantity. [ 32 ]. In addition, the total cellular count obtained by the ACA method used in this study (2 mL) may be less in comparison with the larger volume samples of MFAT from other systems.…”