Development of isolation and cultivation method for outer root sheath cells from human hair follicle and construction of bioartificial skin

Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventio...

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Published in:Biotechnology and bioprocess engineering 2003, 8(2), , pp.151-157
Main Authors: Seo, Young-Kwon, Lee, Doo-Hoon, Shin, Youn-Ho, You, Bo-Young, Lee, Kyung-Mi, Song, Key-Yong, Seo, Seong-Jun, Whang, Sung-Joo, Kim, Young-Jin, Yang, Eun-Kyung, Park, Chang-Seo, Chang, Ih-Seop, Park, Jung-Keug
Format: Article
Language:English
Subjects:
Aging
Cell culture
Culture techniques
Edetic acid
Enzymes
Epithelial cells
Epithelium
Ethylenediaminetetraacetic acids
Follicles
Hair
Keratinocytes
Mitomycin
Mitomycin C
Propagation
Roots
Serum-free medium
Sheaths
Skin
Studies
Tissue engineering
Trypsin
생물공학
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Summary:Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventional primary culture technique to overcome the low propagation and easy aging of epithelial cells during culturing. In a conventional primary culture, the average yield of human ORS cells is 2.1×10^sup 3^ cells/follicle based on direct incubation in a trypsin (0.1%)/EDTA (0.02%) solution for 15 min at 37°C, however, our modified method was able to obtain about 6.9×10^sup 3^ cells/follicle using a two-step enzyme digestion method involving dispase (1.2 U/mL) and a trypsin (0.1%)/EDTA (0.02%) solution. Thus, the yield of primary cultured ORS cells could be increasd three times higher. Furthermore, a total of 2.0×10^sup 7^ cells was obtained in a serum-free medium, while a modified E-medium with mitomycin C-treated feeder cells produced a total of 6.3×10^sup 7^ cells over 17 days when starting with 7.5×10^sup 4^ cells. Finally, we confirmed the effectiveness of our ORS cell isolation method by presenting their ability for reconstructing the bioartificial skin epitheliumin vitro. [PUBLICATION ABSTRACT]
ISSN:1226-8372
1976-3816
DOI:10.1007/BF02940272