Optimum preservation for autologous cultured dermal substitutes.
The application of autologous cultured dermal substitute (CDS) is a promising procedure for improving burn scar contracture. Preparation of CDS requires a period of about 3 weeks before the date of surgery. When the date of surgery is postponed, CDS must be preserved under optimum conditions. This study was thus designed to investigate these conditions. CDS was preserved in culture medium for 2 weeks at 37 degrees C, 25 degrees C, or 4 degrees C in dishes sealed with tape. During this period, culture medium was exchanged every week. CDS fibroblast activity was then examined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell activity was best retained at 37 degrees C, as compared with the other 2 temperatures. The amount of vascular endothelial growth factor (VEGF) released from CDS was measured by enzyme-linked immunosorbent assay. At 37 degrees C, the amount of VEGF was markedly higher. In contrast, the amount of VEGF decreased at the other 2 temperatures. The results of both MTT assay and enzyme-linked immunosorbent assay indicate that preservation of CDS should be carried out at 37 degrees C.
Hayama
Yoshitaka
Y
Department of Plastic and Reconstructive Surgery, Osaka Medical College, Takatsuki city, Japan. pla033@poh.osaka-med.ac.jp
Ueda
Koichi
K
Kuroyanagi
Yoshimitsu
Y
D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media; D020405:Dermis / Q000254:growth & development*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D012879:Skin Physiological Phenomena; D017768:Skin, Artificial; D013696:Temperature; D023822:Tissue Engineering / Q000379:methods*; D014021:Tissue Preservation
D003470:Culture Media