Effect of conditioned medium of vascular endothelial cells on the epithelial-mesenchymal transition of hepatocellular carcinoma cells / 生物医学工程学杂志

This study aims to investigate the effect of substances secreted or metabolized by vascular endothelial cells on epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma cells under indirect co-culture condition. Human hepatocellular carcinoma cell line QGY-7703 was cultured , and then was co-cultured with conditioned medium of human umbilical vein endothelial cells (HUVEC). The morphological changes of QGY-7703 cells were observed by inverted phase contrast microscopy. The migration ability of QGY-7703 cells was analyzed by scratch-wound assays. The effect of conditioned medium on the expression and distribution of EMT related proteins was detected by Western blot and immunofluorescence assays, respectively. The results showed that the QGY-7703 cells gradually changed from polygonal to spindle shape, the migration ability promoted significantly, and both the expression and distribution of EMT related marker changed in a time-dependent manner after co-culturing. The results confirm that vascular endothelial cells can induce EMT in hepatocellular carcinoma cells under indirect co-culture condition.

Indirect Co-Culture of Stem Cells from Human Exfoliated Deciduous Teeth and Oral Cells in a Microfluidic Platform

Oral epithelial-mesenchymal interactions play a key role in tooth development and assist differentiation of dental pulp. Many epithelial and mesenchymal factors in the microenvironment influence dental pulp stem cells to differentiate and regenerate. To investigate the interaction between oral cells during differentiation, we designed a microfluidic device system for indirect co-culture. The system has several advantages, such as consumption of low reagent volume, high-throughput treatment of reagents, and faster mineralization analysis. In this study, stem cells from human exfoliated deciduous teeth were treated with media cultured with human gingival fibroblasts or periodontal ligament stem cells. When human exfoliated deciduous teeth was incubated in media cultured in human gingival fibroblasts and human periodontal ligament stem cells under the concentration gradient constructed by the microfluidic system, no remarkable change in human exfoliated deciduous teeth mineralization efficiency was detected. However, osteoblast gene expression levels in human exfoliated deciduous teeth incubated with human gingival fibroblasts media decreased compared to those in human exfoliated deciduous teeth treated with human periodontal ligament stem cells media, suggesting that indirect co-culture of human exfoliated deciduous with human gingival fibroblasts may inhibit osteogenic cytodifferentiation. This microfluidic culture device allows a co-culture system set-up for sequential treatment with co-culture media and differentiation additives and facilitated the mineralization assay in a micro-culture scale.

Effects of phenytoin on VEGF and SCF in rat bone marrow mesenchymal stem cells and vascular endothelial cells co-culture system / 西安交通大学学报(医学版)

ABSTRACT:Objective To investigate the effects of phenytoin (PHT)on the secretion of vascular endothelial growth factor (VEGF)and stem cell factor (SCF)based on the establishment of indirect co-culture system of rat bone marrow mesenchymal stem cells (BMSCs)and vascular endothelial cells (VECs).Methods Indirect co-culture model of rat BMSCs and VECs was established.Experimental groups:indirect co-culture groups (PHT concentrations were 0,20 and 40 μg/mL);the control group:BMSCs culture group and VECs culture group (PHT concentrations were 0,20 and 40μg/mL).The contents of VEGF and SCF in the culture supernatant were measured using double antibody sandwich ABC-ELISA method on cultivation days 2,4,6.Results ELISA assay of the rBMSCs and rVECs in indirect co-culture supernatants,collected on culture days 2,4 and 6 showed that:① VEGF:On culture day 2,VEGF level in the co-culture groups was significantly higher than those in BMSCs group (P SCF increased as the incubation time increased;but as the incubation time increased, PHT concentration of 40 μg/mL made SCF content decrease.Each group did not significantly differ (P > 0.05 ).Conclusion PHT promotes the secretion of VEGF and may reduce the secretion of SCF.