ZHANG Gui-xue;LIU Yan;HU Peng-fei

;College of Animal Science and Technology, Northeast Agricultural University, Harbin Heilongjiang 150030, PRC

fibroblast;isolation;passage;cryopreservation;histology

The embryonic ages were determined for the best preparation of mouse fibroblasts. Four methods were adapted to verify cryopreservation of mouse fibroblasts. The results showed that embryonic cryopreserving method was best one with 0.86 of thawing viability. The embryos from 13-14 d pregnant mouse were superior to 11-12 d and 15-16 d in isolating, growing, laying and living. The first 6 generations were better than following ones in the same aspects above. Cell laying time became longer and variable time became shorter after the sixth generation. With culture time increasing, fibroblast nuclear size became larger, fibrous filament appeared among fibroblasts, and macrocyst vesicle with floccule appeared in the cells. Cyst vesicle structure with pyknotic granule appeared in 24 h cultured fibroblasts and macrocyst vesicle also appeared in passaging fibroblasts.