CAJ | 학술논문

目的构建适合研究毛囊发育启动的体外培养模型,观察Wnt10b/β连环蛋白信号在毛囊发育初期的表达情况.方法将不同胎龄的SD大鼠胚胎(胎鼠)背部皮肤以明胶海绵为载体,在Dulbecco改良Eagle培养液(DMEM)的气液交界平面中漂浮培养3～6 d,HE染色,观察毛囊发育情况;免疫荧光法动态观察Wnt10b及β连环蛋白在毛囊发育初期的表达;将胎龄15 d胎鼠背部皮肤在含不同胎牛血清浓度的DMEM中培养,观察胎牛血清对毛囊形态发生的影响.结果 (1)胎龄14 d后期至15 d早期的胎鼠背部皮肤培养效果最好,体外培养3 d以内,毛囊的发育保持与体内过程一致,培养3 d后毛囊发育逐渐减慢,直至停止于第4阶段;(2)胎牛血清对毛囊的发育没有明显促进作用;(3)在培养的前几天,Wnt10b/β连环蛋白的表达与体内一致,随着培养天数的增加,Wnt10b/β连环蛋白的表达逐渐减弱,在培养的第6天,毛囊的形态发生停止时,Wnt10b/β连环蛋白的表达也随之消失.结论在体外短期培养的胎鼠背部皮肤毛囊发育与体内基本一致,可以作为研究毛囊发育启动的模型;Wnt10b/β连环蛋白信号的表达与毛囊的早期发育密切相关.
목적 구건괄합연구모낭발육계동적체외배양모형,관찰Wnt10b/β련배단백신호재모낭발육초기적표체정황.방법 장불동태령적SD대서배태(태서)배부피부이명효해면위재체,재Dulbecco개량Eagle배양액(DMEM)적기액교계평면중표부배양3～6 d,HE염색,관찰모낭발육정황;면역형광법동태관찰Wnt10b급β련배단백재모낭발육초기적표체;장태령15 d태서배부피부재함불동태우혈청농도적DMEM중배양,관찰태우혈청대모낭형태발생적영향.결과 (1)태령14 d후기지15 d조기적태서배부피부배양효과최호,체외배양3 d이내,모낭적발육보지여체내과정일치,배양3 d후모낭발육축점감만,직지정지우제4계단;(2)태우혈청대모낭적발육몰유명현촉진작용;(3)재배양적전궤천,Wnt10b/β련배단백적표체여체내일치,수착배양천수적증가,Wnt10b/β련배단백적표체축점감약,재배양적제6천,모낭적형태발생정지시,Wnt10b/β련배단백적표체야수지소실.결론 재체외단기배양적태서배부피부모낭발육여체내기본일치,가이작위연구모낭발육계동적모형;Wnt10b/β련배단백신호적표체여모낭적조기발육밀절상관.
Objective To establish a rat skin organ culture model to study the initiation of hair follicle morphogenesis, and the dynamic expression of Wnt10b/β-catenin in the developing hair follicle. Methods The dorsal skins of SD rat at embryos 14-17 (E14-E17) were cultured on a gelatin sponge-supported tissue culture system at the air/liquid interface of DMEM with 10 % fetal bovine serum (FBS) for 3-6 days, some of which removed from El5 were cultured in DMEM with FBS of different concentrations. HE staining and fluoroimmunoassay were adopted. Results The model we established allowed skin tissues isolated from E14-E15 to develop in a manner that was histologically and temporarily similar to the process in vivo. However, the developing hair follicle ceased to continue when their morphogenetic process reached the forth stage, and the concentration of FBS did not show any significant effect on the development of hair follicle. Expression of Wnt10b/β-catenin was induced in culture, as it also occurred in vivo,but grew weak till it disappeared in culture for 6 days, which was accompanied by development halt of hair follicle. Conclusions A rat skin organ culture model is established in which the morphogenesis of hair follicle takes place in a manner similar to in vivo, and Wnt10b/β-catenin probably has a close connection with the early morphogenesis of hair follicle.