title: Testicular Sertoli cells influence the proliferation and immunogenicity of co-cultured endothelial cells.
abstract: The major problem of the application of endothelial cells (ECs) in transplantation is the lack of proliferation and their immunogenicity. In this study, we co-cultured ECs with Sertoli cells to monitor whether Sertoli cells can influence the proliferation and immunogenicity of co-cultured ECs. Sertoli cells were isolated from adult testicular tissue. ECs were divided into the control group and the experimental group, which included three sub-groups co-cultured with 1 × 10(3), 1 × 10(4) or 1 × 10(5) cell/ml of Sertoli cells. The growth and proliferation of ECs were observed microscopically, and the expression of vascular endothelial growth factor (VEGF) receptor-2 (KDR) was examined by Western blotting. In another experiment, ECs were divided into the control group, the single culture group and the co-culture group with the optimal concentration of Sertoli cells. After INF-γ and TNF-α were added to the culture medium, MHC II antigen expression was detected by immunofluorescence staining and western blotting; interleukin (IL)-6, IL-8 and soluble intercellular adhesion molecule (sICAM) were measured in the culture medium by ELISA. We demonstrated that 1 × 10(4) cell/ml Sertoli cells promoted the proliferation of co-cultured ECs more dramatically than that in other groups (P<0.05). Western blotting showed that 1 × 10(4) cell/ml of the Sertoli cells was most effective in the up-regulation of KDR expression in the co-cultured ECs (P<0.05). Sertoli cells can effectively suppress INF-γ-induced MHC II antigen expression in co-cultured ECs compared with single culture group (P<0.05). TNF-α induced the expression of IL-6, IL-8 and sICAM in ECs. When co-cultured with Sertoli cells, their expressions were significantly lower than in the EC single culture group (P<0.05). ECs co-cultured with Sertoli cells also did not significantly increase the stimulation index of spleen lymphocytes compared to the single culture group (P<0.05). Our results suggested that co-culturing with Sertoli cells can significantly promote the proliferation of ECs, accelerate post-transplant angiogenesis, while reduce EC immunogenicity and stimulus to lymphocytes.
authors.affiliation: Department of Rheumatism and Immunity, The First Affiliated Hospital Xi'an Jiaotong University School of Medicine, Xi'an, Shaanxi 710061, PR China. fanpinggoodluck@163.com
authors.forename: Ping
authors.initials: P
authors.lastname: Fan
authors.forename: Lan
authors.initials: L
authors.lastname: He
authors.forename: Dan
authors.initials: D
authors.lastname: Pu
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Lv
authors.forename: Wenxu
authors.initials: W
authors.lastname: Zhou
authors.forename: Yining
authors.initials: Y
authors.lastname: Sun
authors.forename: Nan
authors.initials: N
authors.lastname: Hu
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000032:analysis; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000379:methods; D042783:Endothelial Cells / Q000166:cytology* / Q000276:immunology* / Q000637:transplantation; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008297:Male; D012708:Sertoli Cells / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000032:analysis
chemical_list: D015415:Biomarkers; D000949:Histocompatibility Antigens Class II; D015850:Interleukin-6; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D040301:Vascular Endothelial Growth Factor Receptor-2