Zheng Yonghua, Chen Xiaoke, Qian Mengjia, Zhang Miaomiao, Zhang Ding, Bai Chunxue, Wang Qun, Wang Xiangdong
Department of Respiratory Medicine, Shanghai Respiratory Research Institute, Zhongshan Hospital, Fudan University School of Medicine, Fenglin Rd, No 180, Xuhui Dist, Shanghai, 320003 China.
Department of thoracic surgery, Shanghai Respiratory Research Institute, Zhongshan Hospital, Fudan University School of Medicine, Fenglin Rd, No 180, Xuhui Dist, Shanghai, 320003 China.
Mol Cell Ther. 2014 Feb 1;2:3. doi: 10.1186/2052-8426-2-3. eCollection 2014.
In the previous studies, telocytes were found near the capillaries in many tissues, especially on the extracellular matrix of blood vessels and positive to CD34 and c-kit. Therefore, the present study aimed to explore if telocytes could produce angiogenesis associated cytokines, promote the proliferation and the angiogenesis of vascular endothelial cells in vitro.
Human lung telocytes were isolated and cultured, and were identified by immunofluorescence cytochemistry with CD34, c-kit and vimentin. Telocytes conditional media (TCM) was prepared, and the expressions of angiogenesis associated cytokines in TCM were detected by ELISA. Human pulmonary microvascular endothelial cells (HPMECs) were cultured with DMEM media or TCM for 72 hours. The proliferation of HPMECs was continuously detected with CCK-8 kit at an interval of 12 hours. HPMECs were also injured by lipopolysaccharide, and cultured with TCM and DMEM respectively, and the tube formation capacity was detected.
Telocytes were positive for CD34, c-kit and vimentin. The expressions of VEGF and EGF in TCM were significantly higher, the proliferation of HPMECs cultured with TCM significantly increased, and the tube formation of HPMECs injured by endotoxin was improved with the culture of TCM, as compared with the culture of DMEM.
The present study provides the evidence that human lung telocytes could produce the growth factors, such as VEGF and EGF. Telocytes conditional media induced the proliferation of pulmonary endothelial cells and prevented from endotoxin-induced compromise of pulmonary endothelial angiogenesis.
在先前的研究中,在许多组织的毛细血管附近发现了端粒细胞,尤其是在血管的细胞外基质上,且对CD34和c-kit呈阳性。因此,本研究旨在探讨端粒细胞是否能产生与血管生成相关的细胞因子,在体外促进血管内皮细胞的增殖和血管生成。
分离并培养人肺端粒细胞,通过用CD34、c-kit和波形蛋白进行免疫荧光细胞化学鉴定。制备端粒细胞条件培养基(TCM),通过ELISA检测TCM中与血管生成相关细胞因子的表达。将人肺微血管内皮细胞(HPMECs)用DMEM培养基或TCM培养72小时。每隔12小时用CCK-8试剂盒连续检测HPMECs的增殖情况。HPMECs也用脂多糖损伤,分别用TCM和DMEM培养,并检测其管形成能力。
端粒细胞对CD34、c-kit和波形蛋白呈阳性。与DMEM培养相比,TCM中VEGF和EGF的表达显著更高,用TCM培养的HPMECs的增殖显著增加,内毒素损伤的HPMECs用TCM培养后其管形成得到改善。
本研究提供了证据表明人肺端粒细胞能产生VEGF和EGF等生长因子。端粒细胞条件培养基诱导肺内皮细胞增殖,并防止内毒素诱导的肺内皮血管生成受损。