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

Pending Publication Date: 2020-11-20
LABREAL BIOTECH KUNMING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a method for culturing rat macrophages, so as to solve the problem that the existing rat macrophage culture method proposed in the above-mentioned background technology has a low purity of macrophages and a slow growth cycle, which is unfavorable for experiments. research question

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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PUM

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Abstract

The invention discloses a rat macrophage culture method. The method comprises the following steps: killing young SD rats which are born for 3 weeks by a neck removal method, soaking the rats in 75% alcohol for disinfection for 3 minutes, transferring the rats into an ultra-clean workbench for operation after disinfection is completed, cutting posterior limb skin, removing leg muscles, exposing thighbones, stripping external muscles of the thighbones cleanly, and cutting off the thighbones; cutting off femoral heads at the two ends of the thighbones, sucking a certain amount of MEM-alpha basalculture medium liquid by using an injector, blowing out bone marrow, and flushing the bone marrow cleanly in the blowing process; blowing and beating the bone marrow for 30 times by using a pasteur pipette to prepare a single-cell suspension, performing centrifuging by using a centrifuge, collecting cells, and discarding the supernatant; and adding erythrocyte lysate. According to the invention, the rat bone marrow is subjected to primary centrifugal separation, erythrocyte lysis, secondary centrifugal separation and culture by using a special culture medium for macrophages, so that the cultured macrophages are relatively high in purity, the culture speed is relative high, and the research requirements are met.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for culturing rat macrophages. Background technique [0002] Macrophages are white blood cells located in tissues derived from monocytes, which in turn are derived from precursor cells in the bone marrow. Both macrophages and monocytes are phagocytic cells that participate in non-specific Heterosexual defense and specific defense, their main function is to phagocytize cell fragments and pathogens in the form of fixed cells or free cells, and activate lymphocytes or other immune cells to make them respond to pathogens. Macrophages are Immune cells have multiple functions and are important objects for the study of cell phagocytosis, cellular immunity and molecular immunology. However, the macrophages cultured by the existing rat macrophage culture methods have low purity and slow growth cycle. It is beneficial to experimental research, so it is necessary to improve t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0786
CPCC12N5/0645C12N2509/00C12N2501/22C12N2501/998
Inventor 范振峰孔令泽李慧贺永胜
Owner LABREAL BIOTECH KUNMING CO LTD
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