Report Case: Cultured Keratinocyte Autograft on Collagen from Amniotic Membrane for Treatment the Injured Human Skin
Cell therapy is an emerging therapeutic strategy aimed at replacing or repairing severely damaged tissues with cultured cells. The epidermis is a major site of self-renewal in which there is constant replacement of cells lost by desquamation in the superficial layers by cell division in the basal layers –epidermal stem cells and transit amplifying cells. These cells are amenable to in vitro expansion and constitute a major area of research in regenerative medicine, tissue engineering, and skin grafting. In this research, cells from patient skin were collected, cultured, amplified in serum free medium. The multilayered structure was made by using insert dishes lined collagen membrane derived from human amniotic membrane and air-lifting. Then, the cells were evaluated by karyotyping and immunohistochemistry. The result shows that the keratinocyte sheet is composed of 4 to 5 layers which the proliferating cells are closely lying on the basal layer. The cells have a nornal karyotype. Keratinocyte sheet is easy to strip off from the flask surface by using collagen as a scaffold. After graft, the result indicates that the diminution of scar in the region which skin is taken off, the relief of pain, the low infection risk, the same effect for split-thickness autograft case.
KeywordsEpidermis epidermal stem cell transit amplifying cell serum free medium amniotic membrane basal layer
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