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
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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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