Screening method for proteinuria component induced renal interstitial fibrosis based on micro-fluidic chip

A microfluidic chip and chip technology, applied in the field of biomedical research, can solve the problems of high cost of treatment, heavy economic burden on individuals and society, and achieve the effect of reducing consumption

Inactive Publication Date: 2013-06-05
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

[0002] Uremia is a major disease that endangers people's lives and health. Its treatment is expensive and brings a heavy economic burden to individuals and society.

Method used

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  • Screening method for proteinuria component induced renal interstitial fibrosis based on micro-fluidic chip
  • Screening method for proteinuria component induced renal interstitial fibrosis based on micro-fluidic chip
  • Screening method for proteinuria component induced renal interstitial fibrosis based on micro-fluidic chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] First carry out HK-2 cell inoculation:

[0038] After human proximal tubular epithelial cells (HK-2) were digested, fresh culture medium was added to adjust the cell density to 1×10 6 / ml, pipette the cell suspension into figure 1 In the central pool of the chip shown, when the cells are observed to flow into the cell culture chamber under the action of the hydrostatic fluid, 5-10ml of fresh culture solution is quickly covered on the surface of the chip, and the HK-2 cells in the chip stop flowing immediately, due to Due to the settling effect of the cells' own gravity, the cells can quickly stop in the cell culture room, put the chip into a 37°C incubator for static culture, take out the chip after about 30min-2h, absorb the excess culture solution on the surface of the chip, and then Then put the chip back into the 37°C incubator to continue culturing.

[0039] Then add the inducing factor: connect the central pool and the syringe pump through the tetrafluoro tube, ...

Embodiment 2

[0043] HK-2 cell inoculation and staining steps are the same as "Example 1"

[0044] Complement C3a is an important component factor in the complement system. According to the existing experimental results in "Example 1", complement C3a is added here for further investigation.

[0045] The specific inducing factor adding process is as follows: connect the central pool and the syringe pump through the tetrafluoro tube, turn on the syringe pump, adjust the flow rate to 0.6 μl / min, and sequentially add the same volume of inducing factors from the stimulant inlet pool: 1- 3 did not add any stimulating factors, as a negative control group; 4-6 respectively added 2.5ng / ml, 5ng / ml, 10ng / ml transforming growth factor (transforming growth factor-β, TGF-β), as a positive control group; 7-9 respectively added the mixture containing different concentrations (1%, 3%, 5%) of ihhs and complement C3a (10nM); 10-12 respectively added 1%, 3%, 5% inactivated healthy human serum ( inactive human...

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Abstract

The invention provides a screening method for proteinuria component induced renal interstitial fibrosis based on a micro-fluidic chip. The micro-fluidic chip comprises twelve identical structure units. The structure units are connected through central pools. Each structure unit is composed of a stimulant inlet pool (1), a fluid buffer channel (2), a cell culture pool (3) and a central pool (4). The screening method for the proteinuria component induced renal interstitial fibrosis based on the micro-fluidic chip integrates chip cell culture, cell and factor interaction, fluid stimulation, cell marking and cell responding signal detection on the chip of several square centimeters, and can be used for analyzing a proteinuria component induced renal interstitial fibrosis mechanism. Compared with a perforated plate technology, the screening method for the proteinuria component induced renal interstitial fibrosis based on the micro-fluidic chip saves the cumbersome step of repeated cell inoculation, solves the problems that different factors cannot act on cells at the same time under the flow condition, reduces the consumption of the cells and reagent, and can achieve a plurality of experiment parameters through one-time operation.

Description

technical field [0001] The invention belongs to the field of applying microfluidic chip technology to biomedical research, and specifically relates to a microfluidic chip-based screening method for renal interstitial fibrosis induced by various proteinuria components. Background technique [0002] Uremia is a major disease that endangers people's lives and health. Its treatment is expensive and brings heavy economic burdens to individuals and society. At present in China, various primary glomerular diseases are still the first cause of uremia, and persistent and unrelieved proteinuria is an independent risk factor for the progression of the above diseases to uremia, a large part of which is due to proteinuria. Urine can damage renal tubular epithelial cells (PTEC) and interstitium, leading to renal interstitial fibrosis. Therefore, it is of great significance to analyze the mechanism of renal interstitial fibrosis caused by various components of proteinuria and to seek supp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12Q1/02
Inventor 秦建华马慧朋
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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