Acellular amniotic membrane: an appropriate scaffold for fibroblast proliferation

Abstract

Various materials have been investigated as possible skin substitutes to repair skin defects such as burns. Because of its unique characteristics, acellular amniotic membrane seems to provide a good scaffold for cell cultures. To investigate the proliferation of fibroblasts on an amniotic membrane scaffold, and the preparation of a temporary skin substitute using this method. Neonatal foreskin tissue was harvested after circumcision and used for isolation of skin fibroblasts. The skin sample was refrigerated in cell-culture solution, and later treated with trypsin, minced, and incubated in the same solution at 37 °C with in an atmosphere of 95% O2 /5% CO2 . The confluent cultures were treated with trypsin, and fibroblasts were subcultured up to the 10th passage. Cells were tested for microbial contamination, presentation of major histocompatibility complex, and karyotype changes. Amniotic membrane was harvested after elective caesarean section from donors who had been screened for infection. The membrane was washed and then subjected to three freeze-thaw cycles, before having the cells removed. The fibroblasts were seeded onto the scaffold, and after 24 h, the prepared skin substitute was ready. This was examined under electron microscopy. The skin substitute showed excellent growth of fibroblasts on the amniotic membrane scaffold. Fibroblasts had excellent adherence to and viability on the acellular amniotic membrane, which seems to provide an acceptable temporary skin substitute that can be used for wounds.