Separation and purification method of mouse intestinal epithelium mucosa lamina propria dendritic cells

A technology of dendritic cells and intestinal epithelium, applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve the problems of LPDC extraction difficulties, achieve good antigen presentation ability, strong operability, The effect of the simple experimental process

Inactive Publication Date: 2012-03-21
梁廷波
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  • Separation and purification method of mouse intestinal epithelium mucosa lamina propria dendritic cells
  • Separation and purification method of mouse intestinal epithelium mucosa lamina propria dendritic cells
  • Separation and purification method of mouse intestinal epithelium mucosa lamina propria dendritic cells

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[0040] In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below with reference to examples. It should be understood that the examples described here are only used to explain the present invention, and are not intended to limit the present invention.

[0041] The invention discloses a method for isolating and purifying dendritic cells in the lamina propria of the intestinal epithelium of mice. Digest into single cells with collagenase. Mononuclear cells were isolated using Ficoll density gradient centrifugation. Finally, CD11c magnetic beads were used to sort the intestinal epithelial mucosa lamina propria dendritic cells. The invention can conveniently, short-term, economically and efficiently obtain a large number of intestinal epithelial mucosa lamina propria dendritic cells with high purity and good activity. Specific steps are as follows:

[0042] ①Obtain mixed cell...

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Abstract

The invention relates to a separation and purification method of mouse intestinal epithelium mucosa lamina propria dendritic cells, which comprises the following specific steps: (1) the acquisition of mixed cells; (2) the acquisition of individual karyocytes: preparing cells acquired after the digestion of collagenase into a suspension, slowly adding into a mouse lymphocyte separation liquid according to a volume ratio of 1:1, and centrifuging at 1800 rpm for 25 minutes; sucking the middle buffy coats, adding into 1-2 milliliters of MACS buffer, and centrifuging at 1200 rpm for 5 minutes; and removing the supernate, and suspending the cells again, wherein the MACS buffer is a PBS (phosphate buffer solution) containing 2mM mol/milliliter EDTA (ethylene diamine tetraacetic acid) and 0.5% FCS (fetal calf serum); and (3) the acquisition of intestinal epithelium mucosa lamina propria dendritic cells. By using the separation and purification method, a large number of high-purity and high-activity intestinal epithelium mucosa lamina propria dendritic cells can be conveniently, economically and efficiently acquired in short time; and the acquired cells keep a good antigen presentation capability under in vitro conditions, thereby providing favorable conditions for the in vitro simulation of in vivo antigen presenting cell induced lymphocyte differentiation.

Description

technical field [0001] The invention relates to the field of separation and purification of cells, and mainly relates to a method for separation and purification of dendritic cells in the lamina propria of mouse intestinal epithelium. Background technique [0002] As early as 1973, American scholar Steinman discovered dendritic cells (Dendritic cells, DC) with powerful antigen presentation function. This type of cell is the professional antigen-presenting cell with the strongest function in the body known so far. It can efficiently take up, process and present antigen. Immature DC has a strong ability to migrate, and mature DC can effectively activate naive T cells. Cells are at the center of initiating, regulating, and maintaining immune responses. [0003] They are usually distributed in small amounts on exposed skin sites, mainly the skin and lining of the nasal cavity, lungs, stomach and intestines. The gut is the largest immune organ in the human body. On the one han...

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Application Information

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IPC IPC(8): C12N5/0784
Inventor 梁廷波张匀陈灵晓白雪莉陈伟
Owner 梁廷波
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