Preparation method of monocyte-derived macrophages

A technology of macrophages and monocytes, applied in the medical field, can solve problems such as unstable results and complicated use, and achieve the effects of simple operation, stable results, and good phagocytic function

Pending Publication Date: 2022-04-15
GANSU UNIV OF CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] However, the existing macrophage preparation methods are complicated to use, and the results are not stable enough, and some require the use of some more special equipment

Method used

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  • Preparation method of monocyte-derived macrophages
  • Preparation method of monocyte-derived macrophages
  • Preparation method of monocyte-derived macrophages

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preparation example Construction

[0022] see Figure 1~3 , in an embodiment of the present invention, a method for preparing monocyte-derived macrophages, the steps of the preparation method are:

[0023] First, draw blood from the cubital vein under sterile conditions, then treat the blood with heparin sodium anticoagulant, and dilute the blood at the same time;

[0024] Add a certain dose of lymphocyte separation solution into a sterile centrifuge tube, and then use a pointed pipette to gently spread the diluted blood on top of the lymphocyte separation solution. The ratio of lymphocyte separation solution to blood is 1:2, and pay attention to the difference between the two. To form a clear interface, perform 2,000 revolutions and 20 minutes of centrifugation;

[0025] After the centrifugation is completed, the centrifuged blood forms four layers according to the density gradient: the uppermost layer is the diluted plasma layer, the middle layer is the mononuclear cell layer, which is in the shape of buffy ...

Embodiment 1

[0029] The GM-CSF is a cytokine for inducing differentiation of hematopoietic cells into macrophages in vitro.

Embodiment 2

[0031] The RPMI-1640 complete medium is a complete medium containing 10 percent FBS.

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Abstract

The invention discloses a preparation method of monocyte-derived macrophages, which comprises the following steps: firstly, extracting elbow venous blood under a sterile condition, then treating the blood with a heparin sodium anticoagulant, and diluting the blood at the same time; adding a certain dose of lymphocyte separation liquid into a sterile centrifugal tube, and then gently spreading the diluted blood on the lymphocyte separation liquid by using a sharp suction tube; the ratio of the lymphocyte separating medium to the blood is 1: 2, and a clear interface is formed between the lymphocyte separating medium and the blood; the method disclosed by the invention is simple and convenient to operate and stable in result, does not need special equipment conditions, and lays a foundation for researching biological functions of macrophages; therefore, a simple, economical and stable method for inducing the monocyte to be differentiated into the macrophage is established, and a good model is provided for future macrophage research. The GM-CSF induced MDM is very similar in the aspects of cell morphology, function and cell surface receptor expression, and the induced MDM can be substituted as a cell model for studying respiratory system diseases.

Description

technical field [0001] The invention relates to the field of medical technology, in particular to a preparation method of monocyte-derived macrophages. Background technique [0002] Pulmonary macrophages are white blood cells located in tissues, derived from monocytes, which are widely distributed in the interstitium of the lung, around the airways of the bronchioles and in the inner corners of the alveolar septa, and some macrophages migrate into the alveoli In the cavity, called alveolar macrophages, which are larger than monocytes, pleomorphic, with pseudopods on the surface, large and irregular nuclei, and varying numbers of lysosomes and phagosomes in the cytoplasm, from Monocytes are regulated by granulocyte-macrophage colony-stimulating factor and other cytokines to stimulate their differentiation and form, and they can survive for one month to their apoptosis. This characteristic also plays a role in maintaining the stability of lung tissue structure and the aseptic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0786C12N5/0789
Inventor 梁丽娟米友军
Owner GANSU UNIV OF CHINESE MEDICINE
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