[An experimental study of cultured keratinocytes on the acellular xenodermis].
OBJECTIVE
To culture the keratinocytes on the acellular pig dermis and establish a composite skin in vitro.


METHODS
Full thickness skin taken from neonatal SD rats of approximate 24-hour-old was cultivated in asepsis condition, which was then separated into epidermal and dermal layers with low-temperature enzyme digestion. The basal lamina cells between the two layers were scraped off and the pure keratinocytes were obtained using gradient density centrifugation. In the experimental group, the keratinocytes were cultured on acellular xenodermis(AX) as the three-dimensional frames. In the control, the keratinocytes were cultured without any trophoblast. The air-liquid interface(ALI) culture continued after the primary culture. Routine histological HE staining and assay of Pancytokeratin and Laminin with strepavidin-biotin-peroxidase complex (SABC) method were used to study the morphology of CK and AX.


RESULTS
HE staining demonstrated the formation of more than 4 lays of keratinocytes and basement membrane, with slight keratinization of the cells. Pancytokeratin(+) in immunohistochemical results confirmed that the cultured cells on AX were keratinocytes, not other kinds of cells. Laminin(+) indicated that new procreant basement membrane appeared in CK.


CONCLUSIONS
Keratinocytes cultured on the acellular pig dermis grow well and form basement membrane. This study implies successful construction of composite skin.
Tan
Qian
Q
Department of Plastic Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
Zhou
Hong-reng
HR
Lin
Zi-hao
ZH
Jiang
Hua
H
D062088:Acellular Dermis; D000818:Animals; D000831:Animals, Newborn; D001485:Basement Membrane / Q000166:cytology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D000078404:Epidermal Cells; D015603:Keratinocytes / Q000166:cytology*; D007633:Keratins / Q000032:analysis; D007797:Laminin / Q000032:analysis; D051381:Rats; D012867:Skin / Q000166:cytology; D016038:Skin Transplantation; D013194:Staining and Labeling / Q000379:methods; D013552:Swine; D014183:Transplantation, Heterologous
D007797:Laminin; D007633:Keratins