Isolated culture method of chicken intestinal tract epithelial GammaDeltaT cells
An intestinal epithelium, isolation and culture technology, applied in the biological field, can solve the problem of not being able to clearly reflect the direct effect of target stimuli, and achieve the effects of reducing the risk of pollution, great innovation and uniqueness, and shortening the time.
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Embodiment 1
[0040] In this example, intestinal epithelial γδT cells were isolated and cultured from 30-day-old 1.5kg healthy white-feathered broiler chickens. Fast for 24 hours first, remove dirt and dander, remove abdomen and leg feathers, and soak for 15 minutes; then soak in 0.1% bromogeramine solution for 10 minutes; take out the broiler chicken, bind it on the operating table, and spray 75% alcohol on the chicken For the abdomen and legs, the abdominal cavity was opened; 1% double-antibody solution was sprayed on the surface of each abdominal organ, and the middle section of the jejunum was found with surgical forceps, and a 3 cm intestinal segment was taken in 1% double-antibody PBS solution.
[0041] Move the solution into the ultra-clean bench, cut the intestinal wall longitudinally, wash it in 1% double-antibody PBS solution three times, transfer it into double-antibody-free PBS solution and wash it twice, peel off the intestinal wall fat and connective tissue with surgical scisso...
Embodiment 2
[0047] ——Method of the present invention and traditional method comparative test
[0048] For comparison experiments, traditional 0.05% collagenase IV digestion was used at 37°C, 5% CO 2 , placed on a shaker and shaken for 45 minutes to promote the complete separation of cells. After density gradient centrifugation and magnetic bead secondary antibody sorting, chicken intestinal epithelial lymphocytes were obtained, and cell viability was detected.
[0049] The contriver uses this method and the method described in comparative example, has all done ten tests and compares, and has carried out the data statistics of cell separation situation, compares as table 1:
[0050] Table 1 Method of the present invention and traditional method separation cell comparative test
[0051]
[0052] After using this method to isolate chicken intestinal epithelial γδT cells, after 48 hours of culture, the cell viability and purity were identified by flow cytometry and data statistics, as sh...
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