摘要
目的:通过建立施万细胞样细胞(SC-L)与幼年大鼠背根神经节(DRG)细胞共培养体系,观察DRG细胞突起的生长情况以及检测脑源性神经营养因子(BDNF)的表达。方法:采用0.1%的I型胶原酶分离培养脂肪源性干细胞(ADSCs)并进行鉴定,然后将其诱导分化为SC-L。分离培养DRG细胞,后将其分为单独培养组(DRG)和共培养组(DRG+SC-L)。DRG组将两份等体积的DRG细胞混合,常规培养;DRG+SC-L组将等体积的DRG细胞与SC-L进行共培养处理。7~10 d后采用苏木素-伊红(HE)染色观察两组DRG细胞的形态,免疫荧光和Western Blot技术检测DRG细胞内BDNF蛋白的表达。结果:与DRG组相比,DRG+SC-L组DRG细胞的数量以及突起的数目和长度明显增加、细胞内BDNF蛋白表达量明显升高(P <0.05)。结论:SC-L可增加DRG细胞的数量以及突起的数目和长度,并可提高细胞内BDNF蛋白的表达。
Objective: To observe the growth of the neurites of dorsal root ganglion( DRG) cells and the expression of brain-derived neurotrophic factor( BDNF) by establishing the co-cultured condition of the Schwann cell-like cells( SC-L) and DRG cells. Methods: The adipose-derived stem cells( ADSCs) were isolated and cultured by using0. 1% collagenase I,then they were identified and induced to differentiate into SC-L. The DRG cells were isolated and cultured,then they were divided into two groups: single culture group( DRG) and co-culture group( DRG + SC-L). In the DRG group,two equal volumes of DRG cells were mixed and cultured regularly;whereas in the DRG + SC-L group,the equal volumes of DRG cells and SC-L were co-cultured. After 7 ~ 10 days,Hematoxylin-eosin staining was used to observe the morphology of DRG cells. The expression of BDNF protein was detected by immunofluorescence and Western Blot. Results: Compared with the cells in the DRG group,the number of the DRG cells,the length and the number of the DRG cells neurites in DRG + SC-L group were increased,and the expression of BDNF was elevated significantly( P < 0. 05). Conclusion: SC-L could increase the number of the DRG cells,the length and the number of the DRG cells neurites and upregulate the expression of BDNF in DRG cells.
作者
任旺
胡朔丹
刘琳
刘茗宇
杜鹏飞
付秀美
Ren Wang;Hu Shuodan;Liu Lin;Liu Mingyu;Du Pengfei;Fu Xiumei(Department of Human Anatomy,Basic Medical College,Chengde Medical University,Chengde 067000,China)
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2020年第6期605-611,共7页
Chinese Journal of Neuroanatomy
基金
河北省教育厅重点项目(ZD2020178)
承德医学院国家级大学生创新创业训练计划项目(2018001)。
