背景:低氧预处理培养的人脂肪细胞间充质干细胞(h-AMSCs)存活率提高的影响因素,例如,涉及BCL2和HSP27表达的凋亡抑制,触发信号表达(VEGF),SCF表达式,OCT-4表达,和CD44+表达。本研究的目的是解释低氧预适应的机制和作用以及低氧预适应暴露的最佳持续时间以提高h-AMSCs的存活率。方法:在h-AMSCs培养物中进行低氧预处理的实验性实验室探索性研究(体外)。本研究分为四个阶段。首先,从患者的脂肪组织中分离h-AMSCs培养物。第二,通过CD44+表型(流式细胞术)表征脂肪组织中的h-AMSCs,在缺氧治疗前的CD90+和CD45-表达。第三,h-AMSCs培养(体外)的低氧预处理在氧浓度为1%的条件下进行24,48和72小时.第四,从h-AMSCs培养的存活观察,检测CD44+的作用,VEGF,SCF,OCT-4,BCL2,HSP27的流式细胞术和凋亡抑制的隧道分析法。结果:回归检验结果显示,时间差对VEGF的表达有影响(p<0.001;β=-0.482),缺氧条件对VEGF的表达也有影响(p<0.001;β=0.774)。路径分析结果显示SCF对OCT-4表达有影响(p<0.001;β=0.985)。回归检验结果显示,对HSP27表达的时间效应(p<0.001;β=0.398)和缺氧预处理也影响HSP27表达(p<0.001;β=0.847)。通路分析显示BCL2表达抑制细胞凋亡(p=0.030;β=-0.442),HSP27表达抑制细胞凋亡(p<0,001;β=-0.487)。结论:h-AMSC培养的低氧预处理已被证明可以增加VEGF的表达,SCF,OCT-4、BCL2和HSP27。这项研究证明并解释了通过VEGF进行低氧预处理(O21%)的培养物中h-AMSC存活率增加的新机制的存在,SCF,OCT-4、BCL2和HSP27。
Background: Contributing factors for improved survival of human adipocytes mesenchymal stem cells (h-AMSCs) cultured through hypoxia preconditioning, in example apoptosis inhibition involving BCL2 and HSP27 expression, trigger signal expression (VEGF),
SCF expression, OCT-4 expression, and CD44+ expression. The objective if this study was to explain the mechanism and role of hypoxic preconditioning and the optimal duration of hypoxic preconditioning exposure to improve survival of h-AMSCs. Methods: An experimental laboratory explorative study ( in vitro) with hypoxic preconditioning in h-AMSCs cultures. This research was conducted through four stages. First, isolation of h-AMSCs culture from adipose tissue of patients. Second, the characterization of h-AMSCs from adipose tissue by phenotype (flowcytometry) through CD44+, CD90+ and CD45-expression before being pre-conditioned for hypoxic treatment. Third, the hypoxic preconditioning in h-AMSCs culture ( in vitro) was performed with an oxygen concentration of 1% for 24, 48 and 72 hours. Fourth, observation of survival from h-AMSCs culture was tested on the role of CD44+, VEGF,
SCF, OCT-4, BCL2, HSP27 with Flowcytometry and apoptotic inhibition by Tunnel Assay method. Results: The result of regression test showed that time difference had an effect on VEGF expression ( p<0.001; β=-0.482) and hypoxia condition also influenced VEGF expression ( p<0.001; β=0.774). The result of path analysis showed that
SCF had effect on OCT-4 expression ( p<0.001; β=0.985). The regression test results showed that time effects on HSP27 expression ( p<0.001; β=0.398) and hypoxia precondition also affects HSP27 expression ( p<0.001; β=0.847). Pathway analysis showed that BCL2 expression inhibited apoptosis ( p=0.030; β=-0.442) and HSP27 expression also inhibited apoptosis ( p<0,001; β=-0.487). Conclusion: Hypoxic preconditioning of h-AMSC culture has proven to increase the expression of VEGF,
SCF, OCT-4, and BCL2 and HSP27. This study demonstrated and explained the existence of a new mechanism of increased h-AMSC survival in cultures with hypoxic preconditioning (O2 1%) via VEGF,
SCF, OCT-4, BCL2, and HSP 27.