Egy értékes hazai baromfifajtánk, a magyar parlagi gyöngytyúk (Numida meleagris) embrionális blasztodermasejtjeinek mély hűtése gén

Abstract

SUMMARY Background: In the last 25 years, advances in the manipulation of the early chick-embryo suggest that cryopreservation of blastodermal cells might offer means to preserve the entire genome of poultry species. The deep-freezing protocol of the embryonic cells of the Hungarian landrace Guinea fowl (Numida meleagris) was elaborated within the framework of the “Elaboration of alterna tive biotechnological methods for development of the Hungarian poultry and rabbit in vitro gene bank” project grant. Objectives: The aim of the study was to conserve the whole genetic material in the form of embryonic cells in the ex situ in vitro Poultry Gene Bank of the Centre for Farm Animal Gene Conservation. Since germline chimeras can be produced with frozen-thawed embryonic cells, the preserved genotype can be obtained immediately in the F1 generation. Materials and Methods: During the experiments the cells were collected at different temperatures (+20°C, +4°C), using two combinations of cryo solutions (10% DMSO, 10% FBS, 80% DMEM and 5% DMSO, 5% EG, 10% FBS, 80% DMEM medium) in two types of cryocontainer (straw and ampoule). The viability of cells was examined with Tripan Blue staining during the deep-freezing procedure. Altogether 8 various protocols were tested. Results and Discussion: Based on our investigations, the collection tempera ture plays a key role in the deep-freezing process of embryonic cells since the cell collection at +4°C increases the proportion of live cells in the frozen-thawed samples. Although, the cryopreservation in ampoules at any cryoprotectant combination and at any cell collection temperature was better than in straws, the differences were not significant. In summary, among the experimental pro tocols the cell collection at +4°C, using 5% DMSO and 5% EG cryoprotectant combination in ampoules is the most efficient method (cell surviving 70.5%) for long-term storage of Guinea fowl embryonic cells.