Repairing agent for promoting wound healing of type 2 diabetes
A technology of accelerators and reagents, applied in the field of diabetes, to achieve the effect of promoting hair follicle growth, promoting hair follicle generation, and promoting wound healing
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Embodiment 1
[0035] Example 1 Establishment of type 2 diabetes mouse model
[0036] (1) Purchase 100 C57 BL / 6 male mice from Beijing Weitong Lihua Experimental Animal Company, about 4 weeks old. After the mice arrive in the mouse room, feed them with ordinary feed for a week to make the mice gradually adapt to the mouse room Environment, let the mice live in an environment with a constant temperature of 25°C, moderate humidity, and alternating light and dark, without controlling the free drinking and eating of the mice;
[0037] (2) After 1 week, they were randomly divided into two groups, 50 rats in the type 2 diabetes group (T2D group), fed with high-sugar and high-fat feed; 50 rats in the normal control group (CHOW group), fed with ordinary basal feed;
[0038] (3) After the mice were fed in groups, the weight changes of the mice were recorded at a fixed time every week. When there was a significant difference in the weight of the two groups of mice, the blood glucose of the mice was me...
Embodiment 2
[0043] Isolation and culture of mouse adipose stem cells and fibroblasts
[0044] (1) Adipose stem cells: C57BL / 6 mice and T2D mice were killed by decapitation, the mice were sterilized with 75% alcohol, the abdomen of the mice was carefully cut open with sterilized scissors and forceps, and the mice were avoided. Hair, in order to avoid adipose tissue contamination, use tweezers to carefully pick up the white adipose tissue around the mouse epididymis, and put it into normal saline immediately;
[0045](2); Fibroblasts: kill the mice within one day after birth, soak them in 75% ethanol for 5 minutes, carefully cut the skin tissue on the back of the mice with sterilized scissors and tweezers, and put them in a double-antibody-containing in saline;
[0046] (3) Put the mouse adipose tissue and skin tissue into the ultra-clean workbench for operation, wash with 0.9% normal saline (containing double antibody) for 3 times, wash off the fat on the surface of the adipose tissue, an...
Embodiment 3
[0049] Example 3 Determination of the difference in expression of NPY in adipose stem cells derived from normal mice and adipose stem cells derived from type 2 diabetic mice
[0050] 1. RNA extraction from adipose-derived stem cells
[0051] (1) Cell extraction: Use a pipette to carefully absorb the culture medium, wash it twice with phosphate buffer, add 600 μl of lysate directly to the culture dish, and carefully pipette with a pipette to fully lyse the cells. Add the material to the DNA removal column; tissue extraction: Weigh approximately 20-30 mg of mouse tissue, put it into a 1.5 ml RNase free centrifuge tube, add 600 µl of lysate, grind it with an electric tissue grinder at high speed for about 1 min, and grind Try to avoid RNA degradation during the process. Put the ground tissue homogenate into a refrigerated centrifuge, centrifuge at 13,000 rpm for 3 min, carefully draw the tissue supernatant with a pipette gun and add it to the DNA removal column;
[0052] (2) Ce...
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