ABSTRACT
BACKGROUND: Endothelial colony-forming cells (ECFCs) transplantation exerts beneficial impact on angiogenesis of impaired tissues caused by ischemia. However, whether this impact is related to the paracrine effect of ECFCs needs to be studied further.OBJECTIVE: To detect the cytokine secretion profile of human umbilical cord blood derived-ECFCs conditioned medium (ECFCs-CM) and explore the effect of ECFCs-CM on the proliferation, migration and tube formation ability of human umbilical vein endothelial cells (HUVECs).METHODS: Human cord blood derived-ECFCs were isolated, cultured in vitro, and then identified based on the previous studies. The cytokines in serum free ECFCs-CM were detected using a cytokines antibody array. HUVECs were cultured with ECFCs-CM, serum free EBM-2 as control. The proliferation, migration and tube formation abilities of HUVECs were examined by cell counting kit-8, scratch test and Matrigel assay, respectively.RESULTS AND CONCLUSION: The cultured cells demonstrated typical characteristics of ECFCs, which showed cobblestone appearance and were positive for CD34, KDR and CD144, but not CD45 or CD133, uptook Dil-acLDL and bond FITC-UEA-I, and tube-like structures were formed on Matrigel. The cytokine antibody array showed that ECFCs-CM significantly upregulated the expressions of 30 kinds of angiogenic factors. Compared with the control group,the HUVECs cultured with ECFCs-CM showed significantly improved proliferation ability at 24, 48 and 72 hours (P <0.01). The migration rate of HUVECs in the experimental group was significantly higher than that in the control group at 12 and 24 hours (P < 0.01). There were more tubular structures in the experimental group than those in the control group (P < 0.05). These results indicate that ECFCs can promote the bioactivity of mature endothelial cells through paracrine action.
ABSTRACT
Objective To examine the effect of endothelial colony-forming cells conditioned medium(ECFCs-CM)on biological function of human dermal fibroblasts(HDFs). Methods Human cord blood derived-ECFCs were isolated and identified based on the previous studies. The cytokines in ECFCs-CM were detected using a cytokines antibody array. HDFs were cultured with ECFCs-CM,using serum free EBM-2 as control. The proliferation of HDFs was examined by Cell Counting Kit-8(CCK-8)and the migration was assessed by scratch test assay. The apoptosis of HDFs was detected by flow cytometry. Results The cells isolated from human cord blood demonstrated typical characteristics of ECFCs. The cytokines antibody array indicated that ECFCs-CM contained large amounts of secreted cytokines such as PDGF-BBand EGF. Compared with the control group,the HDFs cultured with ECFCs-CM showed improved proliferation and migration ability. The number of apoptotic cells was smaller than that of the control group under the environment of serum starvation. Conclusion ECFCs-CM can promote the proliferation and migration of HDFs and inhibit the apoptosis of HDFs under the environment of serum starvation.
