Part of the programme for the conservation of genetic resources of the Czech Republic is the cryopreservation of genetic material, which serves as a source of gene reserves. It is currently used to conserve gene reserves cryopreservation of embryos, sperm and oocytes in the „In Vitro Programme“. However, oocyte cryopreservation has a relatively low success rate due to the size of the cell. The bovine oocyte is approximately 100-120 µm, most of which is cytoplasm. By freezing the karyoplast itself, which contains DNA, the size of the cryopreserved material is significantly reduced. This can increase the likelihood of survival of the genetic material of the oocyte after thawing. The aim of the methodology is therefore to develop and describe an alternative method of cryopreserving biological material from cattle breeding females classified as genetic resources of the Czech Republic.