Testicular Sertoli cells influence the proliferation and immunogenicity of co-cultured endothelial cells
Testicular Sertoli cells influence the proliferation and immunogenicity of co-cultured endothelial cells
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DOI：
10.1016/j.bbrc.2010.12.068
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发表时间：
2011-01-21
影响因子：
3.1
通讯作者：
Hu, Nan
中科院分区：
文献类型：
--
作者：
Fan, Ping;He, Lan;Hu, Nan
内皮细胞（endothelial cells，ECs）在移植中应用的主要问题是缺乏增殖能力和免疫原性。本研究将内皮细胞与支持细胞共培养，观察支持细胞对共培养内皮细胞增殖和免疫原性的影响。从成年睾丸组织中分离支持细胞。将内皮细胞分为对照组和实验组，实验组分为3个亚组，分别与1 × 10（3）、1 × 10（4）和1 × 10（5）个细胞/ml的Sertoli细胞共培养。光镜下观察内皮细胞的生长增殖情况，Western blotting法检测内皮细胞血管内皮生长因子受体2（VEGF receptor-2，KDR）的表达。另一实验将内皮细胞分为对照组、单独培养组和与支持细胞最佳浓度共培养组。加入INF-γ和INF-α后，采用免疫荧光染色和免疫印迹法检测培养液中MHC Ⅱ类抗原的表达，ELISA法检测培养液中白细胞介素（IL）-6、IL-8和可溶性细胞间粘附分子（sICAM）的含量。结果表明，1 × 10（4）cell/ml的Sertoli细胞对共培养的内皮细胞增殖的促进作用显著高于其他各组（P < 0.05）。Western blotting结果显示，1 × 10（4）cells/ml的Sertoli细胞对共培养的内皮细胞KDR表达的上调作用最强（P < 0.05）。与单独培养组相比，Sertoli细胞能有效抑制INF-γ诱导的ECs MHC Ⅱ类抗原表达（P < 0.05）。TNF-α诱导EC表达IL-6、IL-8和sICAM。与Sertoli细胞共培养后，其表达明显低于EC单独培养组（P < 0.05）。与支持细胞共培养的内皮细胞对脾淋巴细胞的刺激指数与单独培养组相比无显著性差异（P> 0.05）。结果表明，支持细胞与内皮细胞共培养可显著促进内皮细胞增殖，促进移植后血管生成，同时降低内皮细胞的免疫原性和对淋巴细胞的刺激性。(C)2010年爱思唯尔公司All rights reserved.
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 x 10(3), 1 x 10(4) or 1 x 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-gamma and INF-alpha 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 x 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 x 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-gamma-induced MHC II antigen expression in co-cultured ECs compared with single culture group (P < 0.05). TNF-alpha 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. (C) 2010 Elsevier Inc. All rights reserved.