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Construction method of in-vitro diabetes mellitus glomerulus model based on micro-fluidic chip

A microfluidic chip and glomerulus technology, applied in the direction of urinary tract/kidney cells, artificial cell constructs, biochemical equipment and methods, etc., to achieve the effect of simple operation and complete functions

Active Publication Date: 2018-04-24
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The precise control of the fluid is closer to the real environment in the body, and the application of microfluidic chip technology and glomerular microtissues to directly establish diabetic nephropathy in vitro is still a blank field

Method used

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  • Construction method of in-vitro diabetes mellitus glomerulus model based on micro-fluidic chip
  • Construction method of in-vitro diabetes mellitus glomerulus model based on micro-fluidic chip
  • Construction method of in-vitro diabetes mellitus glomerulus model based on micro-fluidic chip

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Using the microfluidic chip designed and manufactured by the laboratory, the configuration is as follows: figure 1 shown.

[0038] The chip is composed of a cell inlet pool 1, a collagen inlet pool 2, a medium inlet pool 3, a waste liquid pool 4, a cell culture chamber 5, a medium perfusion chamber 6, a cell ball capture chamber 7, and a cell migration chamber 8;

[0039] The two ends of the cell migration chamber 8 are collagen inlet pools 2, the middle part is in the shape of "Feng", and the horizontal structure in the middle is symmetrically arranged with 7 to 10 cell ball capture grooves 7, and the cell migration chamber 8 passes through the cell ball capture groove 7 on one side It is connected with the medium perfusion chamber 6, and the cell migration chamber 8 is connected with the cell culture chamber 5 through the cell ball capture groove 7 on the other side;

[0040] The cell culture chamber 5 is connected to the cell inlet pool 1 on the upper side and the w...

Embodiment 2

[0045] A method for constructing a diabetic glomerulus model in vitro based on microfluidic chip technology. The specific process is as follows:

[0046] Characterization of primary glomeruli of SD rats and identification of glomerular endothelium and podocytes after inoculation on Matrigel-coated Xiaomin and cultured for 48 hours ( figure 2 ), after perfusing the matrigel collagen into the two collagen channels of the chip, after the Marigel collagen gel, the extracted primary rat glomerulus microtissue cell spheres were made into a suspension, and 10 μl of the suspension was added to the cell inlet pool , and quickly sucked 10 μl of cell culture solution from the waste liquid pool, erected the chip, and moved it into a 37°C incubator to place it; the vertical direction was that the cell culture chamber was facing upwards, and the cell migration chamber was facing downwards; after standing upright for 10 minutes, take it out for observation , the cell spheres are tightly at...

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Abstract

The invention provides a construction method of an in-vitro diabetes mellitus glomerulus model based on a micro-fluidic chip. The micro-fluidic chip is mainly formed by upper-layer and lower-layer PDMS through bonding and sealing, and is composed of a cell inlet tank, a collagen inlet tank, a culture medium inlet tank, a waste liquid tank, a cell culture chamber, a culture medium perfusion chamber, a cell ball capture tank and a cell migration chamber, wherein fluid culture stimulation under the conditions with high-sugar 10-30 mM glucose and 1-5 mM glucose low sugar is added on the chip, perfusion is carried out on a side channel for 48h, so that a glomerulus filtration barrier is formed, perfusion with culture mediums with different sugar concentrations is carried out on a main channel for 96h, and thus the in-vitro diabetes mellitus glomerulus model is established. For the diabetes mellitus glomerulus model based on the micro-fluidic chip, the glomerulus filtration barrier can be established, so that the timed monitoring can be carried out on the glomerular filtration rate, meanwhile, the movement of different constituting cells in the glomeruli can be tracked in real time, andmoreover, the accurate positioning for the initial cell movement can be realized.

Description

technical field [0001] The present invention relates to the technical field of applying microfluidic chip technology to the construction of disease models in vitro, in particular to a method for constructing in vitro diabetic glomerulus models based on microfluidic chips. Background technique [0002] Diabetic nephropathy (diabetic nephropathy, DN) is the main microvascular complication of diabetes, mainly refers to diabetic glomerulosclerosis, a glomerular lesion mainly caused by vascular damage. DN not only brings great pain to the patients themselves and their families, but also brings a heavy economic burden to the family, the medical system and the whole society. Currently, there is no effective treatment for end-stage diabetic nephropathy. Therefore, the key to the clinical treatment of DN is to establish an effective treatment model in the early or non-clinical stage of DN and to reverse renal injury. A large number of DN studies are based on in vitro cell culture o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12N5/071
CPCC12M21/08C12M23/16C12N5/0684C12N2533/54
Inventor 秦建华陶婷婷王丽
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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