Deezagi Abdolkhaleg, Shomali Samira
Department of Molecular Medicine and Biochemistry, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Department of Molecular Medicine and Biochemistry, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Cell J. 2018 Jul;20(2):259-266. doi: 10.22074/cellj.2018.5026. Epub 2018 Mar 18.
Tissue engineering today uses factors that can induce differentiation of mesenchymal stem cells (MSCs) into other cell types. However, the problem of angiogenesis in this differentiated tissue remains an unresolved area of research interest. The aim of this study was to investigate the effects of prostaglandin F-2α (PGF-2α) on the expression of vascular endothelial growth factor (VEGF) in human adipose tissue derived MSCs.
In this experimental research, human adipose tissue was digested using collagenase. The isolated MSCs cells were treated with PGF-2α (up to 5 μg/ml) and incubated for 96 hours. Cell proliferation, secretion of VEGF and cell migration were spontaneously assayed by MTT, BrdU, ELISA, RT-PCR and scratching methods.
Cell growth at 1.0, 2.5, 5 μg/ml of PGF-2α was not significantly reduced compared to control cells, suggesting that these concentrations of PGF-2α are not toxic to cell growth. The results of the BrdU incorporation assay indicated that, in comparison to untreated cells, BrdU incorporation was respectively 1.08, 1.96, 2.0 and 1.8 fold among cells treated with 0.1, 1.0, 2.5 and 5.0 μg/ml of PGF-2α. The scratching test also demonstrated a positive influence on cell proliferation and migration. Cells treated with 1.0 μg/ml of PGF-2α for 12 hours showed the highest relative migration and coverage in comparison to untreated cells. Quantitative VEGF ELISA and RTPCR results indicated an increase in VEGF expression and secretion in the presence of PGF-2α. The amount of VEGF produced in response to 0.1, 1.0, 2.5 and 5.0 μg/ml of PGF-2α was 62.4 ± 3.2 , 66.3 ± 3.7, 53.1 ± 2.6 and 49.0 ± 2.3 pg/ml, respectively, compared to the 35.2 ± 2.1 pg/ml produced by untreated cells.
Stimulation of VEGF secretion by PGF-2α treated MSCs could be useful for the induction of angiogenesis in tissue engineering in vitro.
当今组织工程利用能够诱导间充质干细胞（MSCs）分化为其他细胞类型的因子。然而，这种分化组织中的血管生成问题仍是一个尚未解决的研究热点。本研究的目的是探讨前列腺素F-2α（PGF-2α）对人脂肪组织来源的间充质干细胞中血管内皮生长因子（VEGF）表达的影响。
在本实验研究中，使用胶原酶消化人脂肪组织。将分离出的间充质干细胞用PGF-2α（浓度高达5μg/ml）处理，并孵育96小时。通过MTT、BrdU、ELISA、RT-PCR和划痕法自发检测细胞增殖、VEGF分泌和细胞迁移情况。
与对照细胞相比，1.0、2.5、5μg/ml的PGF-2α处理组细胞生长未显著降低，表明这些浓度的PGF-2α对细胞生长无毒。BrdU掺入试验结果表明，与未处理细胞相比，用0.1、1.0、2.5和5.0μg/ml的PGF-2α处理的细胞中BrdU掺入量分别为未处理细胞的1.08、1.96、2.0和1.8倍。划痕试验也证明了对细胞增殖和迁移有积极影响。与未处理细胞相比，用1.0μg/ml的PGF-2α处理12小时的细胞显示出最高的相对迁移率和覆盖率。定量VEGF ELISA和RTPCR结果表明，在PGF-2α存在的情况下VEGF表达和分泌增加。与未处理细胞产生的35.2±2.1 pg/ml相比，0.1、1.0、2.5和5.0μg/ml的PGF-2α诱导产生的VEGF量分别为62.4±3.2、66.3±3.7、53.1±2.6和49.0±2.3 pg/ml。
PGF-2α处理的间充质干细胞刺激VEGF分泌可能有助于体外组织工程中的血管生成诱导。
