摘要
背景:研究发现,脂肪干细胞胶原复合物可促进脂肪干细胞向成熟脂肪细胞分化、成熟,并可促进血管新生。目的:探查胶原海绵复合脂肪干细胞的生物学特性及其对血管新生的影响。方法:(1)实验组将人脂肪干细胞接种于胶原海绵上,以细胞单独培养为阴性对照组,培养24 h后,检测细胞黏附率;培养第2,4,6天,检测细胞增殖与培养液中血管内皮生长因子水平;(2)鸡胚绒毛尿囊膜实验:暴露孵育7 d的鸡胚尿囊膜,分4组,空白组向尿囊膜加入0.2 mL无菌PBS,脂肪干细胞组加入0.2 mL 2×10~8 L^(-1)第3代脂肪干细胞悬液,胶原海绵组加入胶原海绵,复合组加入2×10~8 L^(-1)第3代脂肪干细胞悬液0.2 mL与胶原海绵。孵育7 d后,测定尿囊膜周围微血管计数。结果与结论:(1)细胞黏附率:脂肪干细胞在胶原海绵上的黏附率为(93.04±0.67)%;(2)细胞增殖与培养液中血管内皮细胞生长因子水平:实验组培养第6天的细胞增殖A值高于对照组(P<0.01),培养第4,6天的血管内皮生长因子水平高于对照组(P<0.05);(3)鸡胚绒毛尿囊膜实验:脂肪干细胞组、胶原海绵组、复合组微血管计数高于空白组(P<0.05),复合组高于脂肪干细胞组、胶原海绵组(P<0.05);(4)结果表明:脂肪干细胞与胶原海绵具有良好的生物相容性,两者复合有良好的促进血管新生作用,其机制可能与促进脂肪干细胞增殖与分泌血管内皮生长因子相关。
BACKGROUND： Studies have found that adipose-derived stem cells （ADSCs）/collagen complexes can promote the ADSCs differentiation and maturation into mature adipocytes and promote angiogenesis.
OBJECTIVE： To explore the biological properties of the ADSCs/collagen sponge composite material and to detect its effect on angiogenesis.
METHODS： （1） ADSCs were cultured on collagen sponge （experimental group） or cultured alone （control group）. After 24 hours of culture, cell adhesive rate of ADSCs was determined with flow cytometry. After 2, 4, 6 days of culture, cell proliferation and level of vascular endothelial growth factor （VEGF） in the culture medium were detected. （2） Chick embryo chorioallantoic membranes were exposed and incubated for 7 days and then divided into four groups： 0.2 mL of sterile PBS was added in the blank group, 0.2 mL of 2×10^8/L passage 3 ADSCs suspension was added in the ADSCs group, collagen sponge was added in the collagen sponge group, and collagen sponge with 0.2 mL of 2×10^8/L passage 3 ADSCs suspension was added in the composite group. After 7 days of incubation, the microvessel count around the chorioallantoic membrane was measured.
RESULTS AND CONCLUSION： （1） The cell adhesive rate of ADSCs to collagen sponge reached to （93.04±0.67）%. （2） The absorbance value （at 6 days of culture） and level of VEGF （at 4 and 6 days of culture） in the experimental group were significantly higher than those in the control group （P 〈 0.01 or P 〈 0.05）. （3） Compared with the blank group, the number of microvessels was significantly higher in the ADSCs, collagen sponge and composite groups （P 〈 0.05）. Moreover, higher amount of microvessels were found in the composite group than the ADSCs and collagen sponge groups （P 〈 0.05）. To conclude, ADSCs can adhere well to the collagen sponge with good biocompatibility and their combined use can improve angiogenesis further by enhancing cell proliferation and VEGF secretion of ADSCs.
出处
《中国组织工程研究》
CAS
北大核心
2017年第22期3531-3535,共5页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(81373646)
康缘基金(HZ1009KY)
江苏省高校优势学科建设项目Ⅱ期(012062003010)
江苏省中医院高峰学术人才培养(y2014cr06)~~
