A simple method for cryopreservation of MDBK cells using trehalose and storage at -80 degrees C.

Madin Darby bovine kidney cells were stored at -80 degrees C using trehalose. Trehalose was loaded into the cells by fluid-phase endocytosis that was facilitated by heat shock at 40 degrees C for 1 h. Loaded cells were gradually frozen and stored at -80 degrees C. Revival of cells was done by quick thawing and immediately seeded in the tissue culture flasks. The membrane integrity of cells was measured at different times post-storage by trypan blue dye exclusion method. It was estimated to be 96.23, 73.84, 57.33, 54.36, 25.47, 50.53 and 46.86% at 0, 7, 60, 90, 120, 160 and 180-day post-storage, respectively. Cryostorage of cells at -80 degrees C may help to reduce the use of liquid nitrogen.