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A kind of microvascular liver chip based on cell aggregates and methods of making and using the same

A technology of microvessels and aggregates, applied in biochemical equipment and methods, methods of supporting/immobilizing microorganisms, tissue cell/virus culture devices, etc., can solve problems such as single structure and few simulated functions, and achieve high Density, to achieve mutual promotion, to achieve the effect of information exchange

Active Publication Date: 2017-09-26
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies of the above-mentioned prior art, the purpose of the present invention is to provide a microvascular liver chip based on cell aggregates and its preparation and use method. The inherent defects of using cells and animal models in drug screening solve the shortcomings of the existing liver cell culture system, such as single structure and few simulated functions, and can effectively carry out the sequential positioning, implantation and culture of multiple cells, and carry out drug discovery. Safety and Efficacy Evaluation

Method used

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  • A kind of microvascular liver chip based on cell aggregates and methods of making and using the same
  • A kind of microvascular liver chip based on cell aggregates and methods of making and using the same
  • A kind of microvascular liver chip based on cell aggregates and methods of making and using the same

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preparation example Construction

[0044] The method for preparing the microvascular liver chip based on cell aggregates includes the following steps:

[0045] (1) Use computer-aided design software (such as Auto-CAD or Solidwork) to design and draw the mask of the organ chip, and accurately draw the microstructure and microchannel graphics of each layer of the chip;

[0046] (2) Through micro-processing technology, especially the method of photolithography, two upper and lower masks are prepared, and chips containing microstructures and microchannels are prepared;

[0047] (3) Align, pressurize, and perform plasma bonding on the upper microvasculature chip, the middle porous membrane and the lower liver organ multicellular co-culture system to form a three-dimensional microstructure and microchannel network;

[0048] (4) Plant different cell types sequentially through the entrance of microvessels and the entrance channel of the culture system, and perform adherent and three-dimensional aggregate culture, and c...

Embodiment 1

[0054] Example 1 Design and Fabrication of Microvascular Liver Chip Based on Cell Aggregates

[0055] Use computer-aided design software (such as Auto-CAD or Solidwork, etc.) to design and draw the microstructure and microchannel graphics of the microvascular liver chip based on cell aggregates. Firstly, the photoresist is spin-coated on the negative, and the photoresist mold is formed after photolithography and self-development. The silicon mold is made by the silicon deep etching technology in micro-electromechanical processing, and the microstructure of the chip can also be processed by the numerical control CNC system. and microchannels. Mix the prepolymer of polydimethylsiloxane (PDMS) with the curing agent, and cast it on the mold of the upper and lower two layers of chips after ultrasonic, stirring, vacuum degassing and other steps, and heat reaction at 70~80℃ for 1~ Heat reaction at 65°C for 3 hours or overnight, then cool naturally. The polydimethylsiloxane (PDMS) c...

Embodiment 2

[0056] Example 2 Cell implantation and localized culture of the liver chip multi-cell co-culture system

[0057] Close the inlet and inlet of the liver multi-cell co-culture system channel, open the inlet and outlet of the microvasculature, through the inlet of the microvasculature, slowly flow the endothelial cell culture solution through the microchannel into the curved vessels of the microvasculature Inside, delicately culture for 2-4 hours, until the endothelial cells adhere to the wall and grow on the porous membrane in the middle to form a vascular endothelial barrier. Close the inlet and outlet of the microvasculature, open the outlet and inlet of the liver multi-cell co-culture system, and slowly inject the culture medium of liver cell aggregates prepared by the hanging drop method into the cavity of the liver multi-cell co-culture system. Relatively small, the culture medium and single cells are easy to pass through, while the diameter of the cell aggregate is relativ...

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Abstract

The invention discloses a micro-vessel liver chip based on cell clusters. The micro-vessel liver chip comprises a micro-vessel system located on the upper layer, a blood-vessel-endothelium-like barrier system in the middle and a liver-organ multi-cell co-culture system on the lower layer, wherein the micro-vessel system and the liver-organ multi-cell co-culture system are arranged on own substrates respectively. The micro-vessel system comprises a bent vessel composed of multiple flow-stopping barriers in a staggered mode, and a micro-vessel inlet and a micro-vessel outlet are formed in the two ends of the bent vessel respectively. The blood-vessel-endothelium-like barrier system is composed of a porous film. The liver-organ multi-cell co-culture system comprises a cell cluster enrichment region and a multi-cell co-culture region, and a culture system inlet and a culture system outlet are formed in the two ends of the liver-organ multi-cell co-culture system respectively. A liver disease model can be established, research on pharmacokinetics and medicine activity can be performed, and the micro-vessel liver chip has the advantages of using a small quantity of samples, achieving medicine low consumption and being portable, economical, efficient and accurate.

Description

technical field [0001] The invention relates to a microvessel liver chip based on cell aggregates and a preparation and use method thereof, belonging to the technical fields of microfluidic chip processing design and biomedicine. Background technique [0002] Developing safe and effective medicines is a long, difficult and expensive process. Cell culture and animal experiments are two experimental platforms widely used and highly relied on in modern medical and pharmaceutical research. Before the drug enters the clinic, in vitro experiments and animal transition experiments are required to test the effects of the drug on in vitro cultured cells and animal bodies, and repeated qualitative and quantitative predictions of pharmacodynamics, pharmacokinetics, and toxicology. However, an important reason for the failure of drug development is that there are unavoidable differences between the models currently used for drug evaluation and human disease models. In general, cell cu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M3/00C12M3/04C12Q1/02
CPCC12M23/16C12M25/06C12M25/14G01N33/5014G01N33/5067
Inventor 顾忠泽郑付印赵远锦付繁繁赵泽魏红梅
Owner SOUTHEAST UNIV
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