Separation method of colonic intraepithelial lymphocytes

Abstract

The invention discloses a separation method of colonic intraepithelial lymphocytes (IEL). The separation method comprises the following steps: carrying out euthanasia on a mouse; pulling out a whole large intestine from an ileocecal region to a rectum position and transversely cutting; putting the intestine into a small culture dish; putting the intestine into 20ml of IEL isolation buffer; pre-heating in a water bath to 37 DEG C and cutting the intestine into small sections with the length of 4mm-5mm; putting the intestine sections into 60ml of the IEL isolation buffer; stirring tissues in the IEL isolation buffer for 30min; filtering intestine extract by a filtering system; washing a beaker by 20ml of RPMI 1640 containing glutamine; filtering a washing solution and sub-packaging IEL extract into two 50ml plastic tubes; and taking an obtained sediment dyed flow antibody. The method disclosed by the invention has the characteristics of convenience in operation, time saving and labor saving, and is suitable for popularization and application.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for separating colonic interepithelial lymphocytes (IEL). Background technique [0002] Intestine intraepitheliallymphocytes (intestineintraepitheliallymphocyteiIEL) are a type of lymphocytes located on the surface of the basolateral membrane between intestinal epithelial cells, which play a very important role in mucosal immune defense and the formation of oral immune tolerance. At present, the separation method of colonic interepithelial lymphocytes in the prior art has relatively cumbersome operation steps and is time-consuming and labor-intensive. Contents of the invention [0003] In order to solve the deficiencies in the prior art, the present invention provides a method for separating lymphocytes between colonic epithelium. The method has the characteristics of convenient operation, time-saving and labor-saving, and is suitable for popularization and app...

AI Technical Summary

Problems solved by technology

At present, the separation method of colonic interepithelial