Description: The essence of the solution according to the invention is a quick and easy method of identifying live, primordial germ cells from all other somatic cells. The method is carried out using a phase-contrast microscope or a microscope with an additional light source in which a light beam passes through cells in a dark field of view. Primordial germ cells in their structure have many organelles, lipids and other granules of storage substances, such as glycogen, after the light beam passes through them, they cause the light beam to scatter, causing the effect of shining of primordial germ cells, while the remaining somatic cells do not glare.
A method of identifying viable, avian primordial germ cells, characterized in that first, fertilized chicken eggs are incubated at a temperature of 37-38.5°C for 48-56 hours until embryos are obtained at stage 13 - 15 according to H&H. In stage 13-15, 2 μl of blood are taken from the peripheral vein, successively taken from the blood or from the cell gonads, placed in 1.5 ml Eppendorf tubes, with 500 μl of culture medium KAv-1 cells, and then the test tubes are centrifuged in a laboratory centrifuge for 3 - 5 min at 800 x g. After centrifugation the medium is replaced with new ones and the cells are pipetted, then 50 μl of the sample are taken from the obtained cell suspension and placed on slide or Petri dish, which are placed under an inverted microscope with an additional light source (bright field of view), then the cell slide is illuminated with white light, and after the light has passed through the slide, the sample is observed and analyzed to identify viable avian primordial germ cells.
Organisation: Bydgoszcz University of Science and Technology
Innovator(s): Agata Szczerba, Marek Bednarczyk, Takashi Kuwana
Category: Agriculture and Aquaculture