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.

中文翻译：

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缺氧环境中脂肪组织间充质干细胞表面标志物的检查

摘要

细胞疗法在许多疾病组中越来越多地用于临床。但是，越来越需要动物实验以及临床前和阶段研究。这项研究的目的是在低氧环境中对某些间充质干细胞（MSC）表面标记的传代进程影响的免疫细胞化学研究。用吸脂方法获得的人脂肪组织的酶促反应收获间质血管级分细胞。细胞培养在25 cm ^2中在1％氧气（O ₂），5％二氧化碳（CO ₂）和94％氮气（N）中于37°C进行使用Dulbecco改良的Eagle培养基（DMEM）以及10％胎牛血清，青霉素-链霉素溶液和L-谷氨酰胺添加剂的烧瓶。表面标记（CD19，CD44，CD90和CD105）以三种方式免疫细胞化学检查：P1，P3和P5。我们的表型分析表明，在组CD90，CD44和CD105中，表达进入MSC的通道P1，P3和P5没有变化。在1％O ₂浓度下，渐进通道中的表面标记没有显着差异。（p = 0.14和p分别为0.55）。缺氧培养基和五个连续传代没有改变MSC表面标记的百分比。氧气浓度是干细胞保养，分化和功能的重要因素。分子氧是体内和体外的信号分子和代谢产物底物。还应检查培养基和补充剂，气体成分和百分比，冷冻和解冻以及几何形状控制。