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BACKGROUND:Stem cels are induced to differentiate into endothelial-like cels that can be used for the treatment of diabetic lower extremity vascular disease. However, it is unclear whether these endothelial-like cels can completely replace endothelial cels to improve vascular disease and what are the differences between endothelial-like cels and endothelial cels. OBJECTIVE:To explore the differences and similarities between endothelial-like cels and human umbilical vein endothelial cels in the aspects of morphology, function, and viability. METHODS:Umbilical cord mesenchymal stem cels and umbilical vein endothelial cels were isolated, cultured and identified using flow cytometry and immunohistochemical method. Isolated umbilical cord mesenchymal stem cels were induced in DMEM-LG/F12 containing 10 μg/L vascular endothelial growth factor, 10 μg/L basic fibroblast growth factor and 2% fetal bovine serum to differentiate into endothelial-like cels folowed by immunohistochemical identification. To compare endothelial-like cels with human umbilical vein endothelial cels, cel migration detection, active substance measurement and three-dimensional angiogenesis test were performed. RESULTS AND CONCLUSION:Isolated umbilical cord mesenchymal stem cels strongly expressed the surface markers of mesenchymal stem cels, and human umbilical vein endothelial cels strongly expressed CD31 and VWF. After induction, the umbilical cord mesenchymal stem cels were identified to highly express CD31 and VWF. Through cel migration, active substance and three-dimensional angiogenesis tests, endothelial-like cels were similar to endothelial cels in the function and activity, and superior to endothelial cels. Cite this article:Hao XJ, Hao HY, Zhu MJ, Yuan Z, Li WW, Chen J, Zhu LY. Endothelial-like cels versus human umbilical vein endothelial cels. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):83-88.
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BACKGROUND:Rat tail colagen type I can promote the increase of muscle fiber cels and migration and tube formation of endothelial cels, which is speculated to provide a more suitable internal environment for the growth of cels. OBJECTIVE:To observe the effect and safety of platelet-derived growth factor-BB (PDGF-BB) with rat rail colagen against apoptosis of human umbilical vein endothelial celsin vitro. METHODS:The passage 4 human umbilical vein endothelial cels were cultured in the medium of rat rail colagen and the reduction ratio in different time points was detected by Alamar Blue. The passage 4 human umbilical vein endothelial cels were divided into four groups and cultured in 24-wel culture plates: control group, PDGF-BB group, H2O2 group, PDGF-BB+colagen group. H2O2 was used to induce cel apoptosis in al the groups. Western blot was used to detect the expression of PDGF-BB, apoptosis-related protein and anti-apoptosis protein after 72 hours. Meanwhile, TUNEL method was used to detect cel apoptotic rate. RESULTS AND CONCLUSION:The tube formation in the human umbilical vein endothelial cels was more than that cultured in normal medium (P < 0.05). Cels cultured with rat tail colagen showed similar growth to normal control cels, indicating rat tail colagen had no cytotoxicity. The expressions of PDGF-BB, Bcl-2, and p-Akt in the PDGF-BB+colagen group were significantly higher than those in the other three groups (P < 0.05), but the expression of Bax was lower than that in the other three groups (P < 0.05). The apoptotic rate in the PDGF-BB+colagen group was lower than the PDGF-BB group and H2O2 group (P < 0.05). These findings indicate that rat tail colagen has no cytotoxicity to human umbilical vein endothelial cells, and rat tail collagen combined with PDGF-BB can predominantly enhance anti-apoptosis effects.
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BACKGROUND:Numerous studies have confirmed that neovascularization plays an important role in the growth, invasion and metastasis of tumors. OBJECTIVE:To investigate the features of CD133+ ovarian cancer stem-like cels differentiating into vascular endothelial cels. METHODS:CD133+ ovarian cancer stem-like cels were successfuly harvested from A2780 ovarian cancer cel lines using serum-free culture method, and incubatedin vitro onto 96-wel plates with or without Matrigel. Then, we observed the capacity of CD133+ ovarian cancer stem-like cels and human umbilical vein endothelial cels to form tube-like structures at different time points. Through xenograft experiments, the role of CD133+ ovarian cancer stem-like cels in the angiogenesis of ovarian cancer was observed using immunofluorescence staining. RESULTS AND CONCLUSION:CD133+ovarian cancer stem-like cels and human umbilical vein endothelial cels cultured with no Matrigel had no corresponding lumen formation, and could not express CD31. But those cultured with Matrigel had lumen formation and expressed CD31 significantly. After tumor formation, human-derived CD31 expression was observed in the tumors. These findings indicate that CD133+ ovarian cancer stem-like cels can differentiate into vascular endothelial cels, and be involved in tumor revascularization.
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BACKGROUND:Drug eluting stents and endothelium stents for clinical treatment of vascular stenosis can lead to delayed endothelialization and restenosis. A rapamycin eluting stent combined with CD34 antibody can play a synergistic role to offset delayed endothelialization and intimal hyperplasia due to antiproliferative drugs, but it is stil in the pilot phase. OBJECTIVE:To observe the ability of rapamycin eluting stent combined with CD34 antibody to capture endothelial progenitor cels, and to observe the differentiation characteristics of the captured cels. METHODS:Scanning electron microscope and indirect immunofluorescence were used to observe the morphology and differentiation characteristics of captured endothelial progenitor cels. Under a fluorescence microscope, we observed the captured endothelial progenitor cels and the degree of endothelialization after implantation of the rapamycin eluting stent combined with CD34 antibody into rabbit ear vein. RESULTS AND CONCLUSION:Under the scanning electron microscope, fusiform-like cels with a diameter of 6-8 μm were captured by the composite stent, and 24 hours later, the cels became ful-shaped. The captured cels had the appearance characteristics of endothelial progenitor cels. Results from indirect immunofluorescence observation showed that there were a lot of red fluorescent spots on the coating which represented adherent cels positive for vascular endothelial growth factor receptor-2; the composite stent was largely covered with vascular endothelial cels at 24 hours after stent implantation, and fuly covered at 48 hours, but there was no abnormal cel cluster. These findings indicate that the rapamycin eluting stent combined with CD34 antibody can be specific to rapidly capture endothelial progenitor cels in the peripheral blood, and the stent can be completely covered with vascular endothelial cels at 48 hours after stent implantation, thereby achieving rapid endothelialization and promoting the repair of endothelial cels.
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BACKGROUND:A new type of nano-hydroxyapatite artificial mechanical heart valve has been developed using pulsed laser deposition technology at the Department of Materials, Hefei University and Anhui Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, China. OBJECTIVE:To investigate the compatibility of nano-hydroxyapatite artificial mechanical heart valve with human umbilical vein endothelial cels. METHODS:Human umbilical vein endothelial cels were in vitroisolated, cultured and passaged to the 2-4 generations, and then the cel suspension was inoculated onto the nano-hydroxyapatite artificial mechanical heart valve. After 3, 7, 12 days of culture, the cel growth on the artificial mechanical heart valve was observed under scanning electron microscope. In addition, the human umbilical vein endothelial cels were respectively cultured in room-temperature and high-temperature extract liquids of nano-hydroxyapatite artificial mechanical heart valve, high-density polyethylene and phenol solution extracts for 72 hours, and then, the proliferation of cels was detected by MTT method. RESULTS AND CONCLUSION:Under the scanning electron microscope, the human umbilical vein endothelial cels were fusiform- or polygon-shaped with protuberances adhered to the artificial mechanical heart value at 3 days of culture; the cels were stretched thoroughly and fused at 7 days of culture; and the cels were confluent to pieces that tightly overlaid the heart valve surface and the extracelular matrix was formed localy at 21 days of culture. Results from MTT test displayed that the nano-hydroxyapatite artificial mechanical heart valve had no cytotoxicity to the human umbilical vein endothelial cels, indicating a good cytocompatibility.