Epidermal morphogenesis in an in-vitro model using a fibroblasts-embedded collagen scaffold.
A novel culture system included a self-designed bi-layer 3-D collagen scaffold with different pore size on both sides and specific culture media for different culture stages. This skin equivalent culture model provides a new investigating system to study the role of extracellular matrix and growth factors including epidermal growth factor (EGF), keratinocyte growth factor (KGF), transforming growth factor beta 1 (TGF-beta1), in the cell-cell and cell-matrix interactions. Keratinocytes were seeded onto the dermal equivalent and incubated under submerged condition for 5 days then proceeding to air-liquid interface cultured either with or without EGF addition. In this study, EGF has a positive effect on the keratinocyte migration and proliferation in the submerged stage. However, when 10 ng per ml of EGF was continual added in the air-lifted stage, a less organized and thin differentiated keratinocyte layers were found. Continual 10 ng per ml of EGF addition in the air-lifted stage resulted in uneven cell-matrix interface, and disorganization of the suprabasal layers. On the contrary, in the air-lifted stage without excess EGF, the epithelium cells will stratify, differentiate, and form an epidermis completed with basal, spinous, granular, and cornified layers. The results showed that time scale modulation of EGF on keratinocyte cell behavior depend on the expression of paracrine or autocrine growth factors (e.g. KGF and TGF-beta1).
Huang
Yi-Chau
YC
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan, ROC.
Wang
Tzu-Wei
TW
Sun
Jui-Sheng
JS
Lin
Feng-Huei
FH
D000818:Animals; D002450:Cell Communication; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured / Q000378:metabolism; D003094:Collagen / Q000737:chemistry*; D003470:Culture Media / Q000378:metabolism; D017931:DNA Primers; D020405:Dermis / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004815:Epidermal Growth Factor / Q000378:metabolism; D004817:Epidermis / Q000378:metabolism* / Q000502:physiology*; D005109:Extracellular Matrix / Q000378:metabolism; D051523:Fibroblast Growth Factor 7 / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D009924:Organ Culture Techniques / Q000379:methods*; D010641:Phenotype; D011108:Polymers; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000378:metabolism / Q000648:ultrastructure; D013552:Swine; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000378:metabolism; D053773:Transforming Growth Factor beta1
D003470:Culture Media; D017931:DNA Primers; D011108:Polymers; D013778:Tetrazolium Salts; D013844:Thiazoles; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D051523:Fibroblast Growth Factor 7; D004815:Epidermal Growth Factor; D003094:Collagen; C022616:thiazolyl blue