title: [Morphological observation of human keratinocytes tridimensionally cocultured with xenogenic acellular dermal matrix].
abstract: 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.
authors: Institute of Burn Research, Southwest Hospital, State Key Laboratory of Trauma, Burns and Combined Injury, Third Military Medical University, Chongqing 400038, China.
authors: Chong-tao
authors: CT
authors: ZHU
authors: Dai-zhi
authors: DZ
authors: PENG
authors: Li-hua
authors: LH
authors: WANG
authors: Bin
authors: B
authors: HE
authors: Bi-xiang
authors: BX
authors: ZHENG
authors: Xin
authors: X
authors: ZHOU
authors: Hai-shui
authors: HS
authors: LUO
authors: Yong
authors: Y
authors: WANG
authors: Jing
authors: J
authors: LIU
mesh_terms: 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