ABSTRACT
This study was undertaken to evaluate whether exosomes from human adipose-derived stem cells (ASC-exo) can stimulate the regeneration of human dermal fibroblasts (HDFs). Immunoblotting and FACS analyses showed that ASC-exo was positive for exosome markers. Fluorescence tracking revealed that the contents of ASC-exo were transferred into the HDFs. ASC-exo treatment also stimulated the proliferation and migration of HDFs in a dose-dependent manner. Similarly, the expression levels of genes involved in skin cell proliferation were increased by ASC-exo. Microarray analysis showed an enrichment of microRNAs that have regenerative function. We suggest that the ASC-exo can stimulate skin cell proliferation.
Subject(s)
Cell Movement , Cell Proliferation , Exosomes , Fibroblasts/physiology , MicroRNAs/analysis , Adipose Tissue/cytology , Cells, Cultured , Humans , Oligonucleotide Array Sequence Analysis , Regeneration , Skin/cytology , Stem CellsABSTRACT
Epstein-Barr virus (EBV) is a herpesvirus associated with lymphomas and carcinomas. While EBV-associated epithelial cell lines are good model systems to investigate the role of EBV in carcinoma, only a few cell lines are available as they are hard to acquire. A greater variety of naturally EBV-infected cell lines which are derived from tumour patients are needed to represent various features of EBVaGC. We characterized cell line YCCEL1, established from a Korean EBVaGC patient, to ascertain whether it can be used to study the roles of EBV in EBVaGC. The expression of EBV genes and cell surface markers was examined by in situ hybridization, RT-PCR, Western blot analysis, immunofluorescence assay and Northern blot analysis. EBV episomal status was analysed by Southern blotting and real-time PCR. This cell line expressed EBV nuclear antigen 1 (EBNA1) and latent membrane protein 2A (LMP2A), but not EBNA2, LMP2B nor LMP1. The majority of the lytic proteins were not detected in YCCEL1 cells either before or after treatment with 12-O-tetradecanoylphorbol-13-acetate. YCCEL1 cells expressed BART microRNAs (miRNAs) at high level but did not express BHRF1 miRNAs. YCCEL1 cells expressed cytokeratin, but not CD21 and CD19, suggesting CD21-independent EBV infection. The latent EBV gene and EBV miRNA expression pattern of YCCEL1 cells closely resembled that of general EBVaGC cases. Our results support the value of YCCEL1 cells as a good model system to study the role of EBV in gastric carcinogenesis.
Subject(s)
Carcinoma/virology , Herpesvirus 4, Human/physiology , Stomach Neoplasms/virology , Blotting, Southern , Cell Line, Tumor , Gene Expression Regulation/physiology , Genome, Viral , Humans , MicroRNAs/genetics , MicroRNAs/metabolism , Real-Time Polymerase Chain Reaction , Receptors, Complement 3d/genetics , Receptors, Complement 3d/metabolism , Viral Proteins/genetics , Viral Proteins/metabolismABSTRACT
BACKGROUND: Previously, we reported that neonatal porcine pancreatic cells transfected with hepatocyte growth factor (HGF) gene in an Epstein-Barr virus (EBV)-based plasmid (pEBVHGF) showed improved proliferation and differentiation compared to those of the control. In this study, we examined if pancreatic cells transfected repeatedly with pEBVHGF can be successfully grafted to control blood glucose in a diabetes mouse model. METHODS: Neonatal porcine pancreatic cells were cultured as a monolayer and were transfected with pEBVHGF every other day for a total of three transfections. The transfected pancreatic cells were re-aggregated and transplanted into kidney capsules of diabetic nude mice or normal nude mice. Blood glucose level and body weight were measured every other day after transplantation. The engraftment of the transplanted cells and differentiation into beta cells were assessed using immunohistochemistry. RESULTS: Re-aggregation of the pancreatic cells before transplantation improved engraftment of the cells and facilitated neovascularization of the graft. Right before transplantation, pancreatic cells that were transfected with pEBVHGF and then re-aggregated showed ductal cell marker expression. However, ductal cells disappeared and the cells underwent fibrosis in a diabetes mouse model two to five weeks after transplantation; these mice also did not show controlled blood glucose levels. Furthermore, pancreatic cells transplanted into nude mice with normal blood glucose showed poor graft survival regardless of the type of transfected plasmid (pCEP4, pHGF, or pEBVHGF). CONCLUSION: For clinical application of transfected neonatal porcine pancreatic cells, further studies are required to develop methods of overcoming the damage for the cells caused by repeated transfection and to re-aggregate them into islet-like structures.
