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本文旨在探讨生理低氧和无血清培养条件下人胎盘基蜕膜间充质干细胞(placental decidua basalis-mesenchymal stem cells,PDB-MSCs)的生物学特征和细胞因子的表达情况。采用密度梯度离心法培养获得PDB-MSCs,利用MTT法、流式细胞术检测PDB-MSCs在不同氧浓度(20%O2和1%O2)、有无血清(10%FBS和0%FBS)条件下各个时间点(6h、12h、24h、48h、72h、96h)的增殖和凋亡情况,并采用酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)检测无血清条件下各个时间点细胞上清液中血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)含量。结果显示,在特定的时间内低氧可以促进PDB-MSCs增殖(P<0.01,n=3),而血清对PDB-MSCs增殖的影响与氧浓度关系密切;同时,在本实验条件下,低氧、无血清分别或联合培养不会导致PDB-MSCs凋亡(P>0.05,n=3);在无血清条件下,24h时低氧组PDB-MSCs表达较高水平的VEGF。以上结果提示,PDB-MSCs可能成为缺血相关组织工程产品种子细胞的一个新来源。
This article aims to investigate the biological characteristics and the expression of cytokines of human placental decidua basalis-mesenchymal stem cells (PDB-MSCs) under hypoxic and serum-free culture conditions. PDB-MSCs were obtained by density gradient centrifugation. MTT assay and flow cytometry were used to detect PDB-MSCs under different oxygen concentrations (20% O2 and 1% O2) and serum-free (10% FBS and 0% FBS) (6h, 12h, 24h, 48h, 72h, 96h) were determined by enzyme-linked immunosorbent assay (ELISA). The cell supernatants Fluid vascular endothelial growth factor (VEGF) content. The results showed that hypoxia promoted the proliferation of PDB-MSCs (P <0.01, n = 3) at a specific time, while the effect of serum on the proliferation of PDB-MSCs was closely related to the concentration of oxygen. At the same time, under the experimental conditions, Oxygen, serum-free or co-culture did not result in apoptosis of PDB-MSCs (P> 0.05, n = 3). Under serum-free conditions, PDB-MSCs expressed higher levels of VEGF at 24h. The above results suggest that PDB-MSCs may be a new source of seed cells in tissue engineering products related to ischemia.