Effects of serum-free culture at the air-liquid interface in a human tissue-engineered skin substitute.
Previous studies have reported that well-defined culture conditions can improve keratinocytes terminal differentiation and reproducibility. The aim of our study was to compare skin substitutes cultured in a complete medium with those cultured in a serum-free medium at the air-liquid interface to optimize the self-assembly method. Skin substitutes, cultured in a serum-free medium over 7, 14, and 21 days, were compared with others cultured in a complete medium (5% serum) over the complete culture period. Masson's Trichrome staining showed that the substitutes cultured in a serum-free medium generated a well-developed and differentiated epidermis. Immunolabeling analyses between the substitutes cultured without serum and those cultured in complete serum showed similar expression of epidermal differentiation markers, dermo-epidermal junction, and dermal extracellular matrix components. On the basis of our Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) results, the skin substitutes cultured in serum-free condition over 21 days of culture at the air-liquid interface showed lower frequencies of the CH(2) symmetric mode of vibrations, which means a better lipid organization of the stratum corneum. No significant difference in hydrocortisone penetration was observed between serum-free medium substitutes and the controls. Results demonstrate that the absence of serum does not compromise the characteristics of the skin substitutes observed in this study.
Centre LOEX de l'Université Laval, Génie tissulaire et régénération: LOEX-Centre de recherche FRSQ du Centre hospitalier affilié universitaire de Québec, Québec, Canada.
Jessica
J
Jean
Geneviève
G
Bernard
Alexandra
A
Duque-Fernandez
François A
FA
Auger
Roxane
R
Pouliot
D000293:Adolescent; D000328:Adult; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D005260:Female; D006801:Humans; D015603:Keratinocytes / Q000166:cytology; D008875:Middle Aged; D012867:Skin / Q000166:cytology*; D017768:Skin, Artificial; D017550:Spectroscopy, Fourier Transform Infrared; D023822:Tissue Engineering / Q000379:methods*; D055815:Young Adult
D016895:Culture Media, Serum-Free