Separation and primary culture method of pigeon craw fibroblasts

Abstract

The invention provides a separation and primary culture method of pigeon craw fibroblasts. The method comprises the following steps of, taking pigeon craw tissues in an embryonic stage, cleaning, shearing the craw tissues into parts with volume of 1 mm < 3 >, uniformly spreading the craw tissues at the bottom of a cell culture dish, adding a small amount of FBS, putting into a cell culture box for6-8 hours, taking out, adding a complete culture medium, continuing culture, replacing the culture medium every 2-3 days, and culturing for 5-7 days to obtain primary cells with the convergence degree of 80-90%. The passage culture comprises the steps of, using PBS preheated to 37 DEG C in advance to clean the primary cells, adding trypsin for digestion, adding a complete culture medium when thecells shrink and turn round under a microscope, stopping digestion, using a pipette for gently blowing and beating the cells, and transferring the cells into a 15 ml centrifugal tube for centrifugation; re-adding the complete culture medium to re-suspend the cells, carrying out passage culture according to a ratio of 1:2, placing in a cell incubator for culture, and regularly replacing the culturemedium. According to the separation and primary culture method, the operation is simple, the obtained cells are large in quantity and high in purity, and subsequent stability culture can be realized.

Description

technical field [0001] The invention relates to the technical field of in vitro culture of poultry crop fibroblasts, in particular to a method for separating and primary culture of pigeon crop fibroblasts. Background technique [0002] The pigeon crop is located in the enlarged part between the esophagus and the gizzard, and has the functions of moistening, storing food, and providing immune protection. The pigeon crop has the ability to synthesize and secrete pigeon milk. During the hatching period, the cell layers of the crop tissue of male and female pigeons begin to proliferate in large numbers, providing guarantee for the formation and accumulation of pigeon milk. However, there are few reports on the isolation and culture of a single type of cell in the current research. Pigeon crop fibroblasts are an important cell group of crop tissue. According to reports, fibroblasts have the ability to synthesize collagen fibrils, fibronectin, and secrete collagen fibers, elastic...

AI Technical Summary

Problems solved by technology

However, the mechanism of rapid proliferation of pigeon crop fibroblasts during the hatching period, the function and differentiation potential of pigeon milk production are still unclear.

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