Technique for Processing and Preservation of Human Amniotic Membrane for Ocular Surface Reconstruction

Human amniotic membrane (HAM) is a useful biological material for the reconstruction of damaged ocular surface. The processing and preservation of HAM is critical to prevent the patients undergoing amniotic membrane transplant (AMT) from cross infections. For HAM preparation human placenta is obtained after an elective cesarean delivery. Before collection, the donor is screened for seronegativity of HCV, Hbs Ag, HIV and Syphilis. After collection, placenta is washed in balanced salt solution (BSS) in sterile environment. Amniotic membrane is then separated from the placenta as well as chorion while keeping the preparation in BSS. Scrapping of HAM is then carried out manually until all the debris is removed and clear transparent membrane is acquired. Nitrocellulose membrane filters are then placed on the stromal side of HAM, cut around the edges with little membrane folded towards other side making it easy to separate during surgery. HAM is finally stored in solution of glycerine and Dulbecco’s Modified Eagle Medium (DMEM) in 1:1 ratio containing antibiotics. The capped borosil vials containing HAM are kept at -80°C until use. This vial is thawed to room temperature and opened under sterile operation theatre conditions at the time of surgery. Keywords—HAM, AMT, ocular transplant