OBJECTIVETo 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.

METHODSSmall scalp specimens were incubated in the presence of dispase at 37 degrees celsius; 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 dermis 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 degrees celsius; 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 alpha-actin antibody.

RESULTSPurified ORS bulge cells, DSCs and DPCs could be harvested from the same human hair follicle.

CONCLUSIONThis new method allows efficient, rapid, and simultaneous isolation and culture of ORS bulge cells, DSCs and DPCs.