METHOD OF INTRODUCING PRIMORDIAL GERM CELLS OF BIRDS INTO AN EMBRYO "IN OVO" Russian patent published in 2024 - IPC C12N5/07 C12N5/10 C12N5/16 A01K67/00 

FIELD: biotechnology.

SUBSTANCE: invention relates to technologies of living systems, in particular to a group of cell technologies for obtaining and introduction of donor embryonic primordial germ cells of birds without transfer of egg contents into a specialized cuvette. Primordial germ cells of birds are obtained by selection and incubation of bird eggs, poultry egg shell is cut circularly, a blood sample is taken for cultivation through a hole in the egg shell under an unopened outer membrane with a glass microneedle from the dorsal aorta of a bird embryo. When collecting the blood sample from the dorsal aorta of the embryo, the needle is inserted parallel to the artery, against the direction of blood flow. Blood samples taken from the embryo are introduced into prepared test tubes with a culture medium and stored in a heating cabinet at temperature of 37 °C until transfer to the culture plate. Primordial germ cells are cultured in a well plate with a culture medium with cell growth factors at temperature of 37 °C for 21 days. Quantitative evaluation of cultures of primordial germ cells is carried out; then, for introduction of primordial germ cells of birds with an edited genome into the recipient embryo, a circular cut of the recipient egg shell is performed. Further, through a hole in the shell of the recipient egg under an unopened membrane using a microinjector with a glass microneedle filled with primordial germ cells, primordial germ cells are injected into a dorsal aorta of a recipient embryo. Needle is inserted parallel to the artery, against the direction of blood flow.

EFFECT: invention provides higher efficiency of processes of obtaining and transfer of primordial germ cells to recipient and obtaining embryos of birds and chickens with introduced donor, possibly genetically modified, primordial germ cells due to significant reduction of embryo injury level, as well as reducing microbial contamination, therefore, increasing the survival rate of embryos, as well as increasing the number of obtained primordial germ cells from one embryo.

2 cl, 1 dwg, 1 ex