人毛囊外根鞘隆起、真皮鞘和毛乳头细胞高效同步分离培养的方法  

Effective isolation and culture of the outer root sheath bulge cells,dermal sheath cells and dermal papilla cells derived from human hair follicle

作　　者：周洪军[1] 胡志奇[1] 谭挺[1] 孙锡金[1] 

机构地区：[1]南方医科大学南方医院整形外科,广东广州510515

出　　处：《南方医科大学学报》2008年第2期193-195,共3页Journal of Southern Medical University

基　　金：广东省科技计划项目(20052070066);广州市科技计划项目(2007Z3-E0021)~~

摘　　要：目的寻求一种快速高效同步分离培养人头皮毛囊外根鞘隆起（Bulge）细胞、真皮鞘细胞和毛乳头细胞的方法。方法将人头皮标本剪成小块,中性蛋白酶中37℃孵育2h后镜下将带外根鞘的毛干从真皮鞘中拔出,组织块法培养外根鞘隆起（Bulge）细胞;从真皮-皮下组织交界处横断头皮,拔出含毛乳头的真皮鞘,胶原酶D37℃孵育6-8h,多次低速离心,毛乳头沉淀重悬后培养,收集的真皮鞘细胞上清液经高速离心后重悬培养;培养的细胞分别用K19或α-平滑肌动蛋白组化鉴定。结果同一标本可获得纯化的外根鞘Bulge细胞,真皮鞘细胞和毛乳头细胞。结论将现有的分离方法改进和综合运用,可以从同一毛囊标本同步高效获取3种成分细胞。Objective To establish an effective method for isolating and culturing outer root sheath （ORS） bulge cells, dermal sheath cells （DSCs） and dermal papilla cells （DPCs） derived from human hair follicle. Methods Small scalp specimens were incubated in the presence of dispase at 37℃ for 2 h, the hair shafts with ORS embedded in the dermal sheath （DS） were extracted under dissecting microscope, and the ORS tissue were inoculated onto Petri dish. The specimens were transected at the interface between the dennis and subcutaneous tissue. The portions of DS and DP （linked with and enclosed by DS） embedded in the adipose tissue were pulled out and incubated with collagenase at 37℃ for 6-8 h, and the DP and DSCs were isolated by repeated low-speed centrifugation and cultured respectively on Petri dishes. The cultured ORS bulge cells were identified by immunohistochemistry with K19 antibody and DPCs and DSCs by immunohistochemistry with α-actin antibody. Results Purified ORS bulge cells, DSCs and DPCs could be harvested from the same human hair follicle. Conclusion This new method allows efficient, rapid, and simultaneous isolation and culture of ORS bulge cells, DSCs and DPCs.

关 键 词：外根鞘隆起 毛乳头 真皮鞘 细胞培养 毛囊 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]