Amniotic membrane preservation in RPMI‐1640 medium – 5 years experience

Purpose: Present amniotic membrane preservation method using RPMI‐1640 medium and brief analysis of performed surgeries. Methods: Amniotic memmbrane preservation and transplantation surgeries in Vilnius University Hospital Center of eye Diseases was started at the end of 2001. Till the June 2007 amniotic membrane was prepared from 5 placentas obtained during the caesarean delivery. The essence of the preparation of the amniotic membrane was as described by H.Dua and other authors. However, because of the supply disturbances in our hospital instead of the traditional mediums we used RPMI‐1640 medium. Before the cuting amniotic membrane to a small pieces we put the whole membrane in RPMI‐1640 medium with high concentration of antibiotics and antifungals and stored in +5ºC temperature for 24 hours. After that amniotic membrane we put on paper cut to small peaces and stored in RPMI‐1640 medium and glicerol in ‐ 80°C. Results: Before the surgery amniotic membrane was warmed in room temperature. No damage of the structure was observed. Hystopathological slides of membrane samples were performed. Preservation medium specimens for microbiological investigation were taken. From all 5 placentas series no growth was observed. Amniotic membrane was used during 179 ocular surface surgeries. Conclusions: 1. RPMI‐1640 medium can be successfully used for amniotic membrane preservation. 2. Before final frosting in ‐ 80°C, 24 hours in high concentration of antibiotics and antifungals can guaranty high impregnation of amniotic membrane with antimicrobial medication. 3. After preservation in RPMI‐ 1640 medium amniotic membrane retains it appearance, clarity and physical durability.