构建人头皮毛乳头细胞和真皮鞘细胞快捷高效分离培养的方法

Effective methods for isolating and culturing human scalp dermal papilla cells and dermal sheath cells rapidly and purely

目的:将显微法和两步酶法相结合,构建快捷分离培养纯的人头皮毛囊毛乳头细胞和真皮鞘细胞的方法。方法:从真皮-皮下组织交界处横断头皮,拔出脂肪组织中的完整毛囊,解剖显微镜下分离出毛乳头和真皮鞘,毛囊毛乳头用2g/L胶原酶37℃孵育4h左右后悬浮培养;真皮鞘碎片接种在涂有胶原的培养板中。培养的细胞用α-平滑肌肌动蛋白组化及碱性磷酸酶染色鉴定及鉴别。结果:获得纯化的毛乳头细胞和真皮鞘细胞。结论:将原有两方法结合后的新技术,既达到了快的目的,也解决了细胞纯度问题,是高效的人毛囊真皮成分细胞分离培养方法。

AIM:To establish the methods by combining microscopy and two- step enzyme me thod so as to isolate and culture human scalp dermal papilla cells(DPCs) and der mal sheath cells(DSCs) rapidly and effectively . METHODS:Small scalp specimens were transected at the interface between dermis and subcutaneous tissue,and then the portions of hair follicle embedded in the adipose tissue were extracted.Dermal papilla(DP) and dermal sheath(DS) were indi vidually isolated from intact hair follicle under dissecting microscope.DP were pre- incubated by 2 g/L collagenase at 37 ℃ for about 4 hours followed by sus pension.The fragments of DS were inoculated on petri dish smeared with collagen. The cultured cells were identified by immunohistochemistry with α - actin anti body and by cytochemistry staining with alkaline phosphatase. RESULTS:DPCs and DSCs could be harvested purely. CONCLUSION:The new technique combing microscopy and two- step enzyme method is of high efficiency in isolating and culturing cells of human hair follicle de rmal portions rapidly and purely.