论文部分内容阅读
目的探讨子痫前期重度患者血清培育后的血管内皮细胞上清是否对人脐带间充质干细胞(UC-MSCs)有趋化作用。方法 2009年10月至11月在中国医科大学附属盛京医院将内皮细胞以1×105个/mL的浓度接种于6孔培养板,进行分组干预:每组用含15%子痫前期重度患者血清(P组)、健康孕妇血清(H组)、不含血清的DMEM培养液(N组)培养24h,取各组上清液于无菌EP管中,离心20min后加入Transwell小室下室,取第5代UC-MSCs用含5%胎牛血清的DF-12调整细胞浓度为1×105个/mL,加入预处理后的Transwell小室上室,将Tran-swell小室置于37℃、5%CO2孵箱孵育24h。取出后4%多聚甲醛固定、伊红染色后于20倍镜下随机记数5个无重叠细胞视野的透膜细胞数,取其平均数代表该组细胞的侵袭力。结果 N组越膜细胞数:40.87±6.33;H组越膜细胞数:45.47±6.89;P组越膜细胞数:56.07±6.76。P组越膜细胞最多,差异有统计学意义(P<0.01,F=20.53)。结论 UC-MSCs具有向子痫前期重度患者血清培育后血管内皮细胞趋化的生理特性。
Objective To investigate whether the vascular endothelial cell supernatant after serum culture in patients with severe preeclampsia has the chemotactic effect on human umbilical cord mesenchymal stem cells (UC-MSCs). Methods Endothelial cells were inoculated into 6-well plates at the concentration of 1 × 105 / mL in Shengjing Hospital, China Medical University from October 2009 to November 2009 for intervention. In each group, 15% of patients with severe preeclampsia Serum (group P), healthy pregnant women (group H) and serum-free DMEM culture medium (group N) were cultured for 24 hours. The supernatant of each group was collected in sterile EP tube and centrifuged for 20 minutes. The fifth generation UC-MSCs were cultured in DF-12 containing 5% fetal bovine serum at a concentration of 1 × 10 5 cells / mL and added to the upper chamber of the pre-treated Transwell chamber. The Tran-swell chamber was placed at 37 ° C. and 5 % CO2 incubator incubated 24h. After removal of 4% paraformaldehyde fixed, after eosin staining at 20 times the number of random counting five non-overlapping cell field of the number of transmembrane cells, taking the average number of representatives of the cell invasiveness. Results The number of cells in group N was 40.87 ± 6.33, the number of cells in group H was 45.47 ± 6.89, and the number of cells in group P was 56.07 ± 6.76. P group more than the membrane cells, the difference was statistically significant (P <0.01, F = 20.53). Conclusion UC-MSCs have the physiological characteristics of chemotaxis of vascular endothelial cells to sera of patients with severe preeclampsia.