Microarray analysis of gene expression in cultured skin substitutes compared with native human skin.
Cultured skin substitutes (CSS), prepared using keratinocytes, fibroblasts, and biopolymers, can facilitate closure of massive burn wounds by increasing the availability of autologous tissue for grafting. But because they contain only two cell types, skin substitutes cannot replace all of the functions of native human skin. To better understand the physiological and molecular differences between CSS and native skin, we undertook a comprehensive analysis of gene expression in native skin, cultured keratinocytes, cultured fibroblasts, and skin substitutes using Affymetrix gene chip microarrays. Hierarchical tree clustering identified six major clusters of coordinately regulated genes, using a list of 1030 genes that were the most differentially expressed between groups. These clusters correspond to biomarker pools representing expression signatures for native skin, fibroblasts, keratinocytes, and cultured skin. The expression analysis revealed that entire clusters of genes were either up- or downregulated upon combination of fibroblasts and keratinocytes in cultured skin grafts. Further, several categories of genes were overexpressed in CSS compared with native skin, including genes associated with hyperproliferative skin or activated keratinocytes. The observed pattern of expression indicates that CSS in vitro, which display a well-differentiated epidermal layer, exhibit a hyperproliferative phenotype similar to wounded native skin.
Smiley
Andrea K
AK
Research Department, Shriners Hospitals for Children, Cincinnati Burns Hospital, Cincinnati, Ohio, USA.
Klingenberg
Jennifer M
JM
Aronow
Bruce J
BJ
Boyce
Steven T
ST
Kitzmiller
W John
WJ
Supp
Dorothy M
DM
D016000:Cluster Analysis; D005819:Genetic Markers; D006801:Humans; D015603:Keratinocytes / Q000502:physiology; D053506:Matrix Metalloproteinase 10; D008666:Metalloendopeptidases / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D012016:Reference Values; D012879:Skin Physiological Phenomena; D017768:Skin, Artificial; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics
D005819:Genetic Markers; D019717:Tissue Inhibitor of Metalloproteinase-3; D008666:Metalloendopeptidases; D053506:Matrix Metalloproteinase 10