| Summary: | 碩士 === 國防醫學院 === 生物及解剖學研究所 === 97 === Tissue-engineered skin substitutes have been proven to be useful for major burn injuries and chronic wound treatment based on clinical practice. A co-cultured skin model has been developed by seeding keratinocytes and fibroblasts on either side of a collagen: PCL biocomposite membrane and the in vivo study showed good engrafting of tissue-engineered skin model on nude mice. In the further development of optimal materials for skin tissue engineering, gelatin/collagen: PCL biocomposites comprising less amount of collagen component were further designed to achieve better cost effectiveness. Comparative cell biocompatibility was shown in the gelatin/collagen: PCL materials relative to the collagen: PCL membranes. Embryonic stem cells can be expanded indefinitely in culture while maintaining their potentials to multiple differentiations. Therefore, induction of keratinocyte differentiation from stem cells may find beneficial for skin tissue engineering. To investigate the effective method of inducing keratinocyte differentiation from stem cells, co-culture of EGFP transfected mice embryonic stem cells, primary human keratinocytes and/or fibroblasts was designed based on gelatin/collagen : PCL biocomposites and Tissue Culture Plastics (TCP) as a control in this work. The results showed that conditions of keratinocyte culture medium and biocomposite membrane help keratinocyte differentiation, respectively. Moreover, combined conditions of keratinocyte culture medium and biocomposite membrane further enhance keratinocyte differentiation from stem cells. On the other hand, co-culture of keratinocytes and stem cells under both TCP and biocomposite membrane conditions may also help keratinocyte differentiation. Co-culture of fibroblasts and stem cells was observed to restrict keratinocyte differentiation in the early stage (in 8 days); however, increasing keratinocyte differentiation was noted after 12 days possibly due to the beneficial microenvironment based on extracellular matrix produced by fibroblasts in the late stage.
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