[Separation and purification of human hair follicle stem cells by micromanipulation and magnetic cell sorting].
OBJECTIVE
To improve the method of obtaining purified and viable human hair follicle stem cells (HFSCs) from bulge cells (BCs).


METHODS
Firstly, the BCs were isolated from human hair follicles by microdissection. Secondly, the CD200+ HFSCs were selected from BCs using magnetic cell sorting method. The viability of these purified HFSCs was detected under light microscope. The purification rate was analyzed by flow cytometry. The pre- and post-purification cells were compared by immunofluorescence staining.


RESULTS
The adherent BCs displayed a typical cobblestone morphology on day 6. The BCs expressed K19 strongly. The viability rate of pre-purification cells was 95.0% +/- 0.6% while that of post- purification cells was 94.2% +/- 1.0%. There was no significant difference (P < 0.05). By flow cytometry and immunofluorescence staining examination, the CD200+ cell rate was 8.31% before cell sorting purification while that was 82.31% after cell sorting purification.


CONCLUSIONS
Highly purified and viable HFSCs could be obtained by micromani pulation and magnetic cell sorting assay.
Tan
Ting
T
Department of Plastic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou Guangdong, 510515, P.R. China.
Hu
Zhiqi
Z
Zhou
Hongjun
H
D000328:Adult; D015703:Antigens, CD / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D018859:Hair Follicle / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D018189:Immunomagnetic Separation; D008297:Male; D008853:Microscopy; D008875:Middle Aged; D012535:Scalp; D013234:Stem Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D055815:Young Adult
D015703:Antigens, CD; C042739:antigens, CD200