Abstract: 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.