title: [Construction of tissue-engineered skin flap in vitro].
abstract: OBJECTIVE
  To construct a tissue-engineered skin flap using composite skin and adipose  tissue   constructed by adipose-derived stem cells(ASCs).
  
  
  METHODS
  Human ASCs isolated from adipose tissue were cultured and identified for their adipogenic, osteogenic and chondrogenic differentiation potentials. ASCs were then mixed with collagen gel for adipogenic induction and observed 15 days later with inverted microscope, oil-red O staining and HE staining. To construct the composite skin, keratinocytes and fibroblasts were isolated from human foreskin. The fibroblasts were mixed with collagen gel and cultured for 5 days, and keratinocytes were seeded on the gel for 4 days before transfer of the culture to air-liquid interface for culture for another 10 days. The adipose tissue and composite skin were then assembled according to the structure of normal skin and cultured for 3 days with HE staining observation.
  
  
  RESULTS
  The cultured ASCs were capable of adipogenic, osteogenic and chondrogenic differentiation, and adipogenic induction of the ASCs-gel complex for 15 days resulted in adipogenic differentiation of the ASCs in gel. The assembled tissue-engineered skin consisted of 3 layers, including a suprabasal layer formed by the stratified and differentiated keratinocytes, the middle layer and sublayer containing numerous cells, and a underlying sublayer formed by the adipogenic ASCs.
  
  
  CONCLUSIONS
  Tissue-engineered skin flap can be constructed by assembling composite skin and adipose derived from cultured keratinocytes, fibroblasts, and ASCs.
authors.affiliation: Department of Plastic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. weichen352@163.com
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.forename: Ping
authors.initials: P
authors.lastname: Jiang
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Chen
authors.forename: Yunjun
authors.initials: Y
authors.lastname: Liao
authors.forename: Jianhua
authors.initials: J
authors.lastname: Gao
mesh_terms: D017667:Adipocytes / Q000166:cytology; D000273:Adipose Tissue / Q000166:cytology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D015603:Keratinocytes / Q000166:cytology; D013234:Stem Cells / Q000166:cytology; D013524:Surgical Flaps; D023822:Tissue Engineering