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3D printing concentration gradient chip for research of model for improving HepG2 insulin resistance by pioglitazone

A technology of insulin resistance and concentration gradient, which is applied in the fields of medicine and engineering, can solve the problems that mass production cannot be realized, and the microfluidic gradient generator is out of the laboratory, etc., and achieve the effect of low reagent consumption

Pending Publication Date: 2022-02-11
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite the huge development potential, the huge and expensive external driving pump makes it only applicable to the laboratory and cannot achieve commercial mass production
The power problem of liquid transportation has become a major obstacle for microfluidic gradient generators to go out of the laboratory

Method used

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  • 3D printing concentration gradient chip for research of model for improving HepG2 insulin resistance by pioglitazone
  • 3D printing concentration gradient chip for research of model for improving HepG2 insulin resistance by pioglitazone
  • 3D printing concentration gradient chip for research of model for improving HepG2 insulin resistance by pioglitazone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Dilute the blue ink stock solution by 200 times, inject 1 ml of blue ink diluent into the first injection port 1 of the chip, and inject 1 ml of deionized water into the second injection port 2 at the same time, keep the pressure on both sides balanced, relying entirely on Capillary action was used to mix and transport the solution, and after a stable colorimetric gradient was formed, ImageJ was used to analyze and process the change in color intensity.

[0030] Such as figure 2 As shown in A, the color gradient of the entire chip is very obvious. At the final outlet, the blue color gradually becomes lighter from left to right, and the far right end is colorless, which confirms the generation of concentration gradient; due to the surface morphology of α-cellulose powder Relatively orderly, the color of the chip is uniform, and there is no obvious coffee ring reaction. processed by ImageJ figure 2 In B, the change of the relative gray value is consistent with the cha...

Embodiment 2

[0032] A method of cell culture in a 3D printed concentration gradient chip for the study of pioglitazone improving HepG2 insulin resistance model is as follows:

[0033] Digest the cells in logarithmic growth phase, count and dilute to 4×10 5 / ml, add the same volume of hydrogel solution to dilute to 2×10 5 / ml, 30 μL of cell suspension per well was inoculated into the circular cell culture pool 4 of the concentration gradient chip, at 37 °C, 5 % CO 2 cultured in an incubator.

[0034] Such as image 3 Shown is the confocal three-dimensional characterization reconstruction of cells cultured in hydrogel. It can be observed that the green fluorescent protein-labeled HepG2 cells are suspended in the middle of the hydrogel medium, and the cells can fully contact each other, and can be passed through The hydrogel transmits signal molecules to distant cells, and the living space of the cells grows exponentially, which is a multi-layer culture method; the cells are evenly distrib...

Embodiment 3

[0036] A method for establishing an insulin-resistant cell model in a 3D-printed concentration gradient chip for the study of pioglitazone improving the HepG2 insulin-resistant model is as follows:

[0037] Digest the cells in the logarithmic growth phase, count and dilute to 4×10 5 / ml, add the same volume of hydrogel solution to dilute to 2×10 5 / ml, 30 μL of cell suspension per well was inoculated into the circular cell culture pool 4 of the concentration gradient chip. After the cells were cultured for 24 h, the medium was aspirated, and a concentration of 1×10 was added to the first injection port 1 of each chip. -5 and 1×10 -7 μmol·L -1 insulin solution. After insulin was co-cultured with the cells for 24 h, 10 μL of the cell culture medium was drawn from each well for ELISA experiments to detect the glucose consumption rate.

[0038] Such as Figure 4 It was shown that under the action of insulin, the level of glucose consumption by cells decreased in a dose-depe...

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Abstract

The invention discloses a 3D printing concentration gradient chip for research of a model for improving HepG2 insulin resistance by pioglitazone. An open-type micro-fluidic chip with a tree-shaped branch structure is designed, diffusion and transportation of fluid are realized by utilizing capillary action of paper-based fibers, the problem that a traditional micro-fluidic gradient generator needs external power is solved, and the gradient generator and a cell three-dimensional culture area are integrated on a same platform, so that high-throughput, repeatable and low-reagent-consumption drug screening experiment operation is realized. An experiment starts with the establishment of an insulin resistance three-dimensional cell model, discusses the reaction degree of human hepatoma carcinoma cell HepG2 insulin resistance induced by different concentrations of insulin and the level of improving HepG2 cell glucose consumption by pioglitazone, and establishes a stable insulin resistance model and a pioglitazone intervention insulin resistance model.

Description

technical field [0001] The invention relates to the fields of medicine and engineering technology, in particular to a 3D printed concentration gradient chip for research on pioglitazone improving HepG2 insulin resistance model. Background technique [0002] Insulin resistance (IR) refers to a pathophysiological state in which the target tissue of the body produces a lower than normal biological effect on insulin at a normal dose. It mainly occurs in the liver and peripheral tissues. The common pathophysiological basis of common clinical diseases such as hypertension, obesity and atherosclerosis. IR is mainly manifested by decreased insulin sensitivity, decreased glucose utilization in peripheral tissues, and liver tissue cannot effectively inhibit gluconeogenesis and glycogenolysis. Diet structure, living habits, genetic and environmental factors can all cause insulin resistance. Usually, researchers study the pathogenesis of IR and screen effective drugs for IR by establi...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12M1/34B01L3/00C12N5/09C12N5/071G01N33/66G01N33/53C12Q1/02
CPCC12M23/16C12M41/46C12N5/0671C12N5/0693G01N33/66G01N33/5044G01N33/53B01L3/502707C12N2513/00C12N2533/78C12N2539/00C12N2501/33B01L2300/12B01L2200/10B01L2200/027G01N2400/00
Inventor 刘爱林刘萌萌刘辉钟瑜雷云
Owner FUJIAN MEDICAL UNIV
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