Application of collagen membrane from amniotic membrane as a substrate for adherent cell culturing in tissue engineering

Abstract

The human amniotic membrane has been widely used in clinical practice to treat all kinds of lesions related to the epithelial surface, corneal epithelial reconstruction, and orthopaedic trauma. Currently, another application aspect of the amniotic membrane of great interest is its use as an in vitro adherent cell culture medium. In this study, the human amniotic membranes were collected under sterile conditions in the operating room and tested negative for HIV, HBV, HCV, and VDRL. The amniotic membranes were then processed to remove the epithelial layer, to obtain a collagen membrane that can be used as a substrate for cell culture adhesion. Following that, the collagen membrane was structurally evaluated by Hematoxylin and eosin (H&E) staining, SEM. Next, we tested the use of collagen membranes as a culture medium for human fibroblasts and evaluated cell adhesion, growth, and development by histological images over time of cell culture. The image results obtained before and after processing showed that the collagen membrane has an acellular structure, maintaining the basement membrane’s structure. Cell culture results exhibited that fibroblasts adhered and developed well on this collagen membrane.