Dairy cow interdigital skin explant model construction method

[0042] (1) Experimental animals

[0043] The hind hooves of the same dairy cow with healthy and good hoof structure newly slaughtered in the slaughterhouse were used as a group of test samples. A total of 6 hind hooves were collected from three cows, and the hoof dirt was treated under the same conditions. The transportation method remained the same throughout the test process. , are transported on ice.

[0044] (2) Construction of cow interdigital skin explant model

[0045] 1. Preparation of culture medium:

[0046] ①Preparation of transport medium: In every 100mL of transport medium, 1 mL of penicillin-streptomycin-amphotericin B solution (100x), 1 mL of L-glutamine solution, 9 μL of 5 μg / mL gentamicin sulfate, and the balance of DMEM / F121:1 medium.

[0047] ② Preparation of washing medium: In every 100 mL of washing medium, 1 mL of penicillin-streptomycin-amphotericin B solution (100x), 4 μL of 2 μg / mL gentamicin sulfate, and the balance is DMEM / F-121:1 culture ba

[0084] 1. Construction method of cow interdigital skin explant model:

[0085] 1) Preparation method of culture medium:

[0086] ①Preparation of transport medium: In every 100mL of transport medium, penicillin-streptomycin-amphotericin B solution (100x) 0.9mL, L-glutamine solution 1.1mL, 6μg / mL gentamicin sulfate 10μL, the rest The amount is DMEM / F121:1 medium.

[0087] ② Preparation of washing medium: In every 100 mL of washing medium, 0.9 mL of penicillin-streptomycin-amphotericin B solution (100x), 6 μL of 3 μg / mL gentamicin sulfate, and the balance of DMEM / F-121:1 culture medium.

[0088] ③ Preparation of tissue culture medium: per 100 mL of tissue culture medium, 0.9 mL of penicillin-streptomycin-amphotericin B solution (100x), 1.1 mL of L-glutamine solution, 10 μL of 6 μg / mL gentamicin sulfate, fetal 11 mL of bovine serum, 66 mL of DMEM high-glucose medium, and the balance of DMEM / F-121:1 medium.

[0089] 2) Sample collection and model assembly: Rinse the co

[0096] 1. Construction method of cow interdigital skin explant model:

[0097] 1) Preparation method of culture medium:

[0098] ①Preparation of transport medium: In every 100mL of transport medium, 1.1mL of penicillin-streptomycin-amphotericin B solution (100x), 0.9mL of L-glutamine solution, 8μL of 4μg / mL gentamicin sulfate, and the rest The amount is DMEM / F121:1 medium.

[0099] ② Preparation of washing medium: In every 100 mL of washing medium, 1.1 mL of penicillin-streptomycin-amphotericin B solution (100x), 2 μL of 1 μg / mL gentamicin sulfate, and the balance of DMEM / F-121:1 culture medium.

[0100] ③ Preparation of tissue culture medium: per 100 mL of tissue culture medium, 1.1 mL of penicillin-streptomycin-amphotericin B solution (100x), 0.9 mL of L-glutamine solution, 10 μL of 4 μg / mL gentamicin sulfate, fetal 9 mL of bovine serum, 63 mL of DMEM high-glucose medium, and the balance of DMEM / F-121:1 medium.

[0101] 2) Sample collection and model assembly: R