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An improved mouse islet cell enrichment method

An islet cell enrichment technology, applied in the biological field, to achieve the effect of ensuring quality, saving time, and good prospects for promotion and application

Active Publication Date: 2021-08-03
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the defects and deficiencies of the existing mouse islet cell enrichment methods, and provide an improved mouse islet cell that can effectively save time, increase the success rate and yield of operation, and maintain the activity of islet cells. Enrichment method

Method used

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  • An improved mouse islet cell enrichment method
  • An improved mouse islet cell enrichment method
  • An improved mouse islet cell enrichment method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Enrichment method of mouse islet cells

[0046] 1. An improved mouse islet cell enrichment method, comprising the enrichment and collection of mouse islets, the steps are as follows:

[0047] (1) Enrichment of mouse islets:

[0048]Incubate the isolated islets with RMPI1640 containing 15% fetal bovine serum in a common incubator at 37°C for 24 hours, set centrifuge parameters at 800RPM, 4°C, and 1min, then centrifuge, discard the supernatant, and resuspend with 5ml RMPI1640 , filter the resuspension with a 0.1mm mesh funnel; invert the funnel upside down into a Petri dish containing 5ml RMPI1640, and then rinse the filter with 5ml RMPI1640. At this time, the white dots can be seen gathered on the petri dish with the naked eye, and the petri dish is rotated clockwise with a radius of about 50 cm and a speed of 10 revolutions per minute on the table to enrich the white dots. Stereo microscope using 10X eyepieces and 4X objective (eg figure 1 ), use a 200 μl m...

Embodiment 2

[0058] Example 2 Example of the use effect of the enriched mouse islet cells

[0059] 1. Take a 25 μl Hamilton syringe, connect the tip of the syringe to a transparent hose with a length of about 15 cm and a diameter of about 0.2 mm, suck the islet tissue from the long pipette tip in Example 1 into the hose, and place it on crushed ice for later use.

[0060] Transplantation of pancreatic islet cells under the renal capsule showed that the transplanted mouse islets were uniformly enriched under the renal capsule ( image 3 ).

[0061] 2. Seven days after the operation, the mouse kidney was surgically removed, and histological paraffin section and immunohistochemical detection were performed on the site of the islet transplantation. The results were as follows: Figure 4 As shown, mouse islet cells were evenly distributed under the capsule, and the immunohistochemical detection of insulin and glucagon suggested that the islet graft had functioned.

[0062] Mouse islet cells i...

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Abstract

The invention discloses an improved mouse islet cell enrichment method. Firstly, after culturing the isolated islets for 6-48 hours, centrifuge the supernatant and resuspend with culture medium, filter the resuspension, and the filter residue is the enriched islet cells; then inject the enriched islet tissue into "U "-shaped tube, using the principle of the connector to gather islet tissue in the "U"-shaped tube. Using this method to enrich mouse islets can significantly improve the success rate and yield of the operation, maintain the activity of islet cells, and effectively save time; this method is a simpler, faster, cheaper and more efficient method for enriching mouse islet cells , has practical application value in the fields of mouse islet transplantation model construction, new drug experiment, basic endocrinology research, etc., and has a good prospect of promotion and application.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, it relates to an improved mouse islet cell enrichment method. Background technique [0002] At present, organ transplantation is one of the best ways to treat various end-stage diseases, but rejection after transplantation is still the most important reason affecting the long-term survival of patients after transplantation, and a lot of research in this field still needs to be carried out. In the study of transplantation immunity, appropriate animal models are essential; currently commonly used animal models for transplantation immunity research include skin transplantation, heart transplantation, and islet transplantation, among which mouse islet transplantation is relatively simple because of its relatively simple operation, Due to the advantages of low price and accurate observation index, it has become the most widely used transplantation immune model at present. [0003] Mou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0676C12N2509/00
Inventor 马毅章旭之李仲秋邓荣海吴旭坤饶佳伟周晓转
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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