Enzyme digestion to isolate and culture human scalp dermal papilla cells: a more efficient method.
In this study, we show a more efficient method for isolation and cultivation of dermal papilla cells from hair follicles of human scalp skin. The dermal partments of low hair follicles were pulled out from cutaneous fat and the bulb epithelium was teased out from the fibrous sheath with attached dermal papilla by applying gentle pressure with the tip of an occal forceps. When these fibrous sheaths were entirely digested into isolated cells by collagenase D but the dermal papillae were justly to be digested, collagenase D was discarded and the dermal papillae were isolated completely out from the resuspension solution by repeated low-speed centrifugation and transferred to another dish for free-floating culture. This procedure markedly simplifies the steps of isolated dermal papilla operation and relieves the laborious tension. Furthermore, dermal papillae could be isolated on a large-scale and remained intact. After collagenase digestion, the dermal papillae showed very high adherent rate and quicker growth than that of microdissection, which suggests that the definition factor of dermal papilla cell migration was relaxed and some structure had been activated or exposed. The cells exhibited a multi-layer forming property and spread-out growth style. They showed positive with alcian blue, with toluidine blue O for different gradient pH and PAS, which was similar to the staining results of in situ dermal papilla. It suggests that the culture papilla cells still synthesize and excrete neutral and acid mucopolysaccharides. Our results demonstrate that the papilla cells in culture condition still remain the ability to synthesize the specific extracellular matrix components of in situ dermal papilla, which supports the concept that the dermal papilla cell, a highly specialized fibroblast, especially is involved in hair growth regulation.
Wu
Jinjin J
JJ
Department of Dermatology, Daping Hospital, The Third Military Medical University, Chongqing 400042, People's Republic of China. wjjjj@163.com
Liu
Rongqing Q
RQ
Lu
Yuangang G
YG
Zhu
Tangyou Y
TY
Cheng
Bo
B
Men
Xue
X
D000293:Adolescent; D000328:Adult; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D018859:Hair Follicle / Q000166:cytology* / Q000378:metabolism; D006651:Histocytochemistry; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D012535:Scalp / Q000166:cytology*; D013194:Staining and Labeling
D016326:Extracellular Matrix Proteins; D017364:Collagenases