AI Summary & Key Takeaways
AI InsightConclusion: Histologically, an ex vivo produced oral mucosa equivalent (EVPOME) that consisted of a stratified epidermis on a dermal matrix was successfully developed by the method used.
📌 3 Key Takeaways
Abstract
The aim of this study was to produce of ex vivo oral mucosal equivalent. The continuous stratified layer of human oral keratinocytes was grown on a cadaveric human dermal matrix in a defined culture medium without a feeder layer. Materials and methods: Human oral keratinocytes from a keratinized oral mucosa biopsy specimen were enzymatically dissociated and cultured in a serum-free defined culture medium, "EpiLife" (Cascade Biologics, Portland, OR, USA), containing a low calcium concentration of 0.06 mM. Oral keratinocytes were expanded one passage. Once a sufficient population of keratinocytes was reached, they were seeded onto the type IV collagen coated acellular nonimmunogenic cadaveric human dermis (AlloDerm), at cell densities of 1.8 × 105. To form a continuous epithelial monolayer, and to enhance keratinocyte differentiation, oral keratinocyte-AlloDerm composites were cultured while submerged in a high calcium concentration of 1.2 mM medium for 4 days. After 4 days, AlloDerm-keratinocyte composites were raised to an air-liquid interface to encourage stratification of the epithelial monolayer. After additional 7 days, they were taken for histologic examination at 11 days postseeding of the keratinocytes on the AlloDerms. Results: Histologically, on day 11, EVPOME's development showed multilayered epithelium comprising basal, suprabasal, and parakeratinized layers. Conclusion: Histologically, an ex vivo produced oral mucosa equivalent (EVPOME) that consisted of a stratified epidermis on a dermal matrix was successfully developed by the method used.
Özet
The aim of this study was to produce of ex vivo oral mucosal equivalent. The continuous stratified layer of human oral keratinocytes was grown on a cadaveric human dermal matrix in a defined culture medium without a feeder layer. Materials and methods: Human oral keratinocytes from a keratinized oral mucosa biopsy specimen were enzymatically dissociated and cultured in a serum-free defined culture medium, "EpiLife" (Cascade Biologics, Portland, OR, USA), containing a low calcium concentration of 0.06 mM. Oral keratinocytes were expanded one passage. Once a sufficient population of keratinocytes was reached, they were seeded onto the type IV collagen coated acellular nonimmunogenic cadaveric human dermis (AlloDerm), at cell densities of 1.8 × 105. To form a continuous epithelial monolayer, and to enhance keratinocyte differentiation, oral keratinocyte-AlloDerm composites were cultured while submerged in a high calcium concentration of 1.2 mM medium for 4 days. After 4 days, AlloDerm-keratinocyte composites were raised to an air-liquid interface to encourage stratification of the epithelial monolayer. After additional 7 days, they were taken for histologic examination at 11 days postseeding of the keratinocytes on the AlloDerms. Results: Histologically, on day 11, EVPOME's development showed multilayered epithelium comprising basal, suprabasal, and parakeratinized layers. Conclusion: Histologically, an ex vivo produced oral mucosa equivalent (EVPOME) that consisted of a stratified epidermis on a dermal matrix was successfully developed by the method used.