Slide climbing method of PGCs (primordial germ cells)

Abstract

The invention discloses a slide climbing method of PGCs (primordial germ cells) and belongs to the biotechnology field. The slide climbing method comprises the steps as follows: purifying the PGCs; washing the purified PGCs with a PBS (phosphate buffer solution) for three times, and centrifugally collecting the cells; selecting and putting 100-200 cells in PBS droplets on a glass slide under a microscope for washing for one time; selecting and putting the cells in a paraformaldehyde solution with the concentration of 4% on the glass slide for washing for one time; selecting and putting the cells on paraformaldehyde droplets with the concentration of 4% on the glass slide for fixation for 10-30 min; and removing the paraformaldehyde with the concentration of 4% through adsorption, and finishing slide climbing of the cells while fixing the cells. According to the slide climbing method, the problem that the purified PGCs are difficult to climb the glass slide is solved, and the time and the cost are saved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for climbing primordial germ cells. Background technique [0002] Primordial germ cells (PGCs) are precursor cells that form germ cells, which can specifically generate sperm and eggs in sexually mature individuals (Durcova-Hills G et al., PloS one, 2008.3: e3531). As an oviparous animal, poultry is difficult to obtain a single embryo, so the research on PGCs cells is particularly important. With the establishment of the PGCs cell in vitro culture system, the research on modern conservation technology and transgenic technology based on PGCs cells has developed rapidly. [0003] PGCs cells are an ideal raw material for preparing transgenic animals. In 2006, Marie-Cecile van de Lavoir and others successfully established the long-term culture technology of chicken PGCs cells in vitro. The PGCs cells cultured in vitro for about 200 days still have the characteristic...

AI Technical Summary

Problems solved by technology

[0006] The existing primordial germ cell climbing method is as follows: put the glass slide in a petri dish, add feeder cells, after the feeder cells grow into a single layer on the glass slide, use mitomycin or γ-ray After the treatment terminates its division, add PGCs cells, after about 24 hours of cultivation, PGCs cells are adsorbed on the feeder layer cells, and the cell climbing is completed, take out the slides, wash with PBS, and fix them with 4% paraformaldehyde or other The liquid is fixed and the subsequent experiment is carried out. The whole process takes more than 2 days, which is time-consuming and laborious.

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