口腔颌面外科杂志2024，Vol.34Issue(3)：170-179,10.DOI:10.12439/kqhm.1005-4979.2024.03.002
含毛喉素的条件培养液诱导脂肪来源干细胞神经分化的体外实验研究
Forskolin-containing conditioned medium induces neural differentiation of adipose-derived stem cells:An in vitro experimental study
摘要
Abstract
Objective:To explore the neural differentiation of adipose-derived stem cells(ASCs)induced by neural-induced conditional medium in vitro,and to study the optimal concentration of forskolin and the effect of optimal concentration on neurogenic differentiation of ASCs in vitro.Methods:ASCs were isolated by enzyme digestion method.The phenotype and differentiation ability of stem cells are identified by flow cytometry and cell staining.Nerve induction differentiation was performed in vitro by neural-induced conditional medium containing different concentrations of forskolin,and the optimal concentration was screened by immunofluorescence staining.ASCs were induced by neural-induced conditional medium containing the optimal concentration of forskolin and the morphological changes of cells were observed by microscope before induction,6 h after induction and 24 h after induction.Effects of induction on rat ASCs were analyzed by real-time quantitative polymerase chain reaction(RT-qPCR)and immunofluorescence.Results:The optimal concentration of forskolin in neural-induced conditional medium was 10 µmol/L,and the cells induced by neural-induced conditional medium containing 10 µmol/L forskolin could rapidly induce ASCs to differentiate in the direction of nerve formation.After induction,the cell morphology was similar to nerve cells.Results of RT-qPCR and immunofluorescence indicated that the expressions of anti-class Ⅲβ-tubulin(Tuj1),neurofilament-M(NF-M),nestin and S100 β at the mRNA level and protein level were increased after induction(P<0.05).Conclusion:Nerve differentiation of ASCs can be induced by conditional medium containing 10 µmol/L forskolin,and the induced cells are expected to become seed cells of facial nerve conduits.关键词
脂肪来源干细胞/细胞分化/神经再生/毛喉素Key words
adipose-derived stem cells/cell differentiation/nerve regeneration/forskolin分类
医药卫生引用本文复制引用
刘旭凌,胡颖涵,孙家悦,朱泽宇,陆家瑜..含毛喉素的条件培养液诱导脂肪来源干细胞神经分化的体外实验研究[J].口腔颌面外科杂志,2024,34(3):170-179,10.基金项目
国家自然科学基金(82071160、82370933) （82071160、82370933）
上海市高水平地方高校创新团队(SSMU-ZDCX20180900) （SSMU-ZDCX20180900）
