Examination of Adipose Tissue-derived Mesenchymal Stem Cell Surface Markers in a Hypoxic Environment
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xamination of Adipose Tissue-Derived Mesenchymal Stem Cell Surface Markers in a Hypoxic Environment Gulsemin Çiçeka, *, Emine Utlu Ozenb, Fatma Oz Bagcıc, Selcuk Dumanc, T. Murad Aktanc, and Ayse Ozlem Gundeslioglud aKanuni
Sultan Suleyman Research and Education Hospital, Histology and Embryology Dep., Istanbul, Turkey b Afyon Public Hospital, Histology and Embryology Dep., Afyon, Turkey c Necmettin Erbakan University, Meram Faculty of Medicine, Histology and Embryology Dep., Konya, Turkey d Kent Hospital, Plastic reconstructive and aesthetic surgery Dep., Izmır, Turkey *e-mail: [email protected] Received March 12, 2020; revised March 27, 2020; accepted March 27, 2020
Abstract—Cellular therapies are increasingly used clinically in many disease groups. However, animal experimentation and preclinical and phase studies are increasingly needed. The aim of this study is immunocytochemical investigation of the effect of passage progression on some mesenchymal stem cell (MSC) surface markers in a hypoxic environment. Stromal vascular fraction cells were harvested with an enzymatic reaction of human adipose tissue obtained with a liposuction procedure. Cell cultivation was performed at 37°C in 1% oxygen (O2), 5% carbon dioxide (CO2), and 94% nitrogen (N), in 25 cm2 flasks using Dulbecco’s Modified Eagle Medium (DMEM) with additives of 10% fetal bovine serum, Penicillin-Streptomycin solution, and L-glutamine. Surface markers (CD19, CD44, CD90, and CD105) were examined immunocytochemically in three passages: P1, P3, and P5. Our phenotypic analysis showed that in groups CD90, CD44, and CD105 surface expressions were unchanged in the passages P1, P3, and P5 that progressed to MSC. There was no significant difference in the surface markers in the progressive passages in 1% O2 concentration. (p = 0.14 and p = 0.55, respectively). Hypoxic media and five serial passages did not change the percentage of MSC surface markers. Oxygen concentration is an important factor for the care, differentiation, and function of stem cells. Molecular oxygen is the signal molecule and metabolite substrate both in vivo and in vitro. Culture media and supplements, gas composition and percentages, freezing and thawing as well as geometric shape control should also be examined. Keywords: stem cell, hypoxia, cell culture, immunocytochemistry DOI: 10.1134/S1990519X20050028
INTRODUCTION Mesenchymal stem cells (MSCs) are multipotent stem cells that can differentiate into a variety of cell types (adipocyte, osteocyte, chondrocyte, myocyte, and neuron-like cells), and can easily be isolated and multiplied in vitro (Liu et al., 2007; Ashjian et al., 2003). They can be isolated from bone marrow, fat tissue, the umbilical cord, the placenta, the synovium, the endometrium, menstrual blood, fetal liver, muscle tissue, and tooth pulp (Nancarrow-Lei et al, 2003). Stromal vascular fraction (SVF) is a heterogeneous mixture in which cells are isolated by enzymatic or nonenzymatic separation of adipose tissue. Stem cells, endothelial cells, erythrocytes, fibrob
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