shelf-life evaluation of bilayered human skin equivalent, myderm？保质期评价双层人类皮肤等效,myderm .pdfVIP

Shelf-Life Evaluation of Bilayered Human Skin Equivalent, MyDermTM 1 1 1 1,2 1 Wan Tai Seet , Manira Maarof , Khairoji Khairul Anuar , Kien-Hui Chua , Abdul Wahab Ahmad Irfan , Min Hwei Ng1, Bin Saim Aminuddin1,3, Bt Hj Idrus Ruszymah1,2* 1Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia, 2 Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia, 3 Ear, Nose and Throat Consultant Clinic, Ampang Puteri Specialist Hospital, Selangor, Malaysia Abstract Skin plays an important role in defense against infection and other harmful biological agents. Due to its fragile structure, skin can be easily damaged by heat, chemicals, traumatic injuries and diseases. An autologous bilayered human skin equivalent, MyDermTM, was engineered to provide a living skin substitute to treat critical skin loss. However, one of the disadvantages of living skin substitute is its short shelf-life, hence limiting its distribution worldwide. The aim of this study was to evaluate the shelf-life of MyDermTM through assessment of cell morphology, cell viability, population doubling time and functional gene expression levels before transplantation. Skin samples were digested with 0.6% Collagenase Type I followed by epithelial cells dissociation with TrypLE Select. Dermal fibroblasts and keratinocytes were culture-expanded to obtain sufficient cells for MyDermTM construction. MyDermTM was constructed with plasma-fibrin as temporary biomaterial and evaluated at 0, 24, 48 and 72 hours after storage at 4uC for its shelf-life determination. The morphology of skin cells derived from MyDe