Rat macrophage culture method
A macrophage and culture method technology, applied in cell dissociation method, cell culture active agent, tissue culture and other directions, can solve the problems of slow growth cycle, low macrophage purity, unfavorable experimental research, etc., to achieve improved purity, The effect of shortening the cultivation period and high utilization rate
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Embodiment 1
[0021] Rat macrophage culture method, the method comprises the following steps:
[0022] S1: Young SD rats born 3 weeks old were killed by neck dislocation, and disinfected by soaking them in 75% alcohol for 3 minutes. After the disinfection was completed, they were transferred to an ultra-clean workbench for operation. The skin of the hind limbs was cut, the muscles of the legs were removed, and the femur was exposed. , peel off the external muscles of the femur, and cut off the femur;
[0023] S2: Cut off the femoral head at both ends of the femur, use a 5ml syringe to absorb the MEM-α basal medium, blow out the bone marrow, and rinse the bone marrow during the blowing process;
[0024] S3: Use a Pasteur pipette to blow the bone marrow 30 times to make a single-cell suspension, centrifuge through a centrifuge, collect the cells, and discard the supernatant;
[0025] S4: Add 3ml of erythrocyte lysate, lyse at 4°C for 3min, then centrifuge in a centrifuge, collect the cells, ...
Embodiment 2
[0033] Rat macrophage culture method, the method comprises the following steps:
[0034] S1: Young SD rats born 3 weeks old were killed by neck dislocation, and disinfected by soaking them in 75% alcohol for 3 minutes. After the disinfection was completed, they were transferred to an ultra-clean workbench for operation. The skin of the hind limbs was cut, the muscles of the legs were removed, and the femur was exposed. , peel off the external muscles of the femur, and cut off the femur;
[0035] S2: Cut off the femoral head at both ends of the femur, use a 10ml syringe to absorb the MEM-α basal medium, blow out the bone marrow, and rinse the bone marrow during the blowing process;
[0036] S3: Use a Pasteur pipette to blow the bone marrow 30 times to make a single-cell suspension, centrifuge through a centrifuge, collect the cells, and discard the supernatant;
[0037] S4: Add 6ml red blood cell lysate, lyse at 4°C for 3 minutes, then centrifuge to collect the cells, discard ...
Embodiment 3
[0045] Rat macrophage culture method, the method comprises the following steps:
[0046] S1: Young SD rats born 3 weeks old were killed by neck dislocation, and disinfected by soaking them in 75% alcohol for 3 minutes. After the disinfection was completed, they were transferred to an ultra-clean workbench for operation. The skin of the hind limbs was cut, the muscles of the legs were removed, and the femur was exposed. , peel off the external muscles of the femur, and cut off the femur;
[0047] S2: Cut off the femoral head at both ends of the femur, use a 15ml syringe to absorb the MEM-α basal medium, blow out the bone marrow, and rinse the bone marrow during the blowing process;
[0048]S3: Use a Pasteur pipette to blow the bone marrow 30 times to make a single-cell suspension, centrifuge through a centrifuge, collect the cells, and discard the supernatant;
[0049] S4: Add 9ml of erythrocyte lysate, lyse at 4°C for 3 minutes, then centrifuge to collect the cells, discard t...
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