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

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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Problems solved by technology

So it leads to many difficulties in LPDC extraction

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