[Morphological observation of human keratinocytes tridimensionally cocultured with xenogenic acellular dermal matrix].
OBJECTIVE
To establish the tridimensional culture method for tissue-engineered skin to observe the histomorphological change in human immortal KC strain (HacaT)cocultured with xenogenic acellular dermal matrix (ADM).


METHODS
The ADM was prepared from SD rats by a modified method. HaCaTs were cultured in defined KC-serum free medium. HaCaTs in log growth phase were inoculated on ADM at the cell density of 2 x 10(5)/cm(2). They were submergedly cultured for 5 days and then changed to air-liquid phase culture for another 5 days. ADM and growth of HaCaTs on day 1 and 5 after cocultured with ADM were observed with scanning electron microscope. The histological change in ADM and HaCaTs on day 1, 5, and 10 after cocultured with ADM were examined by HE staining.


RESULTS
The gross appearance of ADM was white with smooth and soft texture, and intact collagen bundles without cellular residue. HaCaTs adhered and stretched out pseudopodia on the surface of the ADM on day 1 after combined culture, and a monolayer of cells was formed on day 5, growing into 3-6 layers of cells on day 10 with a tendency to grow into ADM.


CONCLUSIONS
SD rats ADM is benefit for the adhesion of HaCaTs and the permeation of nutrient solution, from which an engineered multiple-layered human skin can be obtained within 10 days.
ZHU
Chong-tao
CT
Institute of Burn Research, Southwest Hospital, State Key Laboratory of Trauma, Burns and Combined Injury, Third Military Medical University, Chongqing 400038, China.
PENG
Dai-zhi
DZ
WANG
Li-hua
LH
HE
Bin
B
ZHENG
Bi-xiang
BX
ZHOU
Xin
X
LUO
Hai-shui
HS
WANG
Yong
Y
LIU
Jing
J
D000818:Animals; D002478:Cells, Cultured; D018920:Coculture Techniques; D020405:Dermis / Q000166:cytology*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017768:Skin, Artificial; D023822:Tissue Engineering / Q000379:methods