论文部分内容阅读
目的探讨地黄低聚糖(RGO)对分离、培养的人脂肪组织源性间充质干细胞(ADMSCs)分泌血管内皮细胞生长因子(VEGF)的影响。方法人脂肪组织用2.5 g/L胶原蛋白酶Ⅰ消化、分离ADMSCs。取沉淀的细胞进行培养,用免疫细胞化学染色法鉴定其表面CD44、CD105、CD45和CD34分子的表达。以IMDM培养基作为空白对照,加入RGO配成不同浓度(0、1、10、100及400 mg/L)的IMDM分别培养ADMSCs,用MTT比色法检测ADMSCs的增殖。培养72 h后,用ELISA法测定不同培养基中VEGF的含量。结果免疫细胞化学染色显示,分离培养的细胞表面CD44+、CD105+、CD34-、CD45-。MTT比色法的结果显示,与空白对照组相比,1和10 mg/L组无明显差别,100及400 mg/L组明显升高(均P<0.01),且100 mg/L组明显高于400 mg/L组(P<0.05)。0、1、10、100、400 mg/L5个组VEGF的含量,分别为(627.88±33.86)、(655.50±40.29)、(666.50±43.20)、(843.50±53.05)和(722.88±51.34)ng/L。结论RGO对ADMSCs的增殖有促进作用,且呈一定的浓度依赖性,并导致其分泌VEGF的作用增强。
Objective To investigate the effect of Resveratrol oligosaccharide (RGO) on the secretion of vascular endothelial growth factor (VEGF) from human adipose tissue-derived mesenchymal stem cells (ADMSCs) isolated and cultured. Methods Human adipose tissue was digested with 2.5 g/L collagenase I and ADMSCs were isolated. Precipitated cells were cultured and their surface expressions of CD44, CD105, CD45, and CD34 molecules were identified by immunocytochemical staining. IMDM medium was used as a blank control. ADMSCs were cultured with different concentrations (0, 1, 10, 100 and 400 mg/L) of IMDM cultured with RGO. The proliferation of ADMSCs was detected by MTT assay. After 72 hours of culture, the VEGF content in different culture media was determined by ELISA. Results Immunocytochemical staining showed that the cultured cells had CD44+, CD105+, CD34-, and CD45-. The results of MTT colorimetry showed that there was no significant difference between the 1 and 10 mg/L groups and the 100 and 400 mg/L groups (all P<0.01), and the 100 mg/L group was significantly different from the blank control group. Higher than the 400 mg/L group (P < 0.05). The levels of VEGF in 0, 1, 10, 100, and 400 mg/L groups were (627.88±33.86), (655.50±40.29), (666.50±43.20), (843.50±53.05), and (722.88±51.34) ng, respectively. /L. Conclusion RGO promotes the proliferation of ADMSCs, and it is in a concentration-dependent manner, which leads to its enhanced secretion of VEGF.