Two-layer cell culture system organ chip and preparation method thereof

A cell culture and organ-on-a-chip technology, applied in the field of biomedicine, can solve problems such as difficulty in miniaturization and integration, single use, and time-consuming, and achieve the effects of high parallel culture capacity, small dosage, and simple equipment

Inactive Publication Date: 2014-08-13
SOUTHEAST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Technical problem: The purpose of the present invention is to provide a two-layer cell culture system organ chip, which can carry out parallel three-dimensional culture of multi-cells and interact with drugs, and evaluate the safety and effectiveness of drugs, which solves the problem of traditional drug screening. Animal models are expensive, time-consuming, and ethically controversial, which solves the shortcomings of the existing cell culture system, such as cell adherent growth, single use, and difficulty in miniaturization and integration.

Method used

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  • Two-layer cell culture system organ chip and preparation method thereof
  • Two-layer cell culture system organ chip and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The preparation of embodiment 1 chip

[0029] 1) Preparation of polymethacrylic acid (PMMA) chips

[0030] Computer-aided design software was used to design and draw the microstructure and microchannel pattern of the three-layer chip of the two-layer cell culture organ chip, and the microstructure and microchannel of the three-layer polymethacrylic acid (PMMA) chip were fabricated by numerical control CNC system. Wash each layer of chips with tap water and distilled water respectively, and wipe the fingerprints, oil stains and other stains remaining on the chip surface with ethanol. On the double-sided adhesive film, the microstructures and microchannels required for the preparation of the chip are processed by a lettering machine, and the three-layer chip is aligned, glued, and pressure-stitched to make four inlets, four cell culture chambers, and two upper and lower layers. A co-culture system, a detection cell with two layers of dialysis membranes, and a two-layer c...

Embodiment 2

[0033] Example 2 Preparation of microporous membrane two-layer co-culture system chip

[0034] Use computer-aided design software to design and draw the microstructure and microchannel patterns of the upper and lower layers of the two-layer cell culture organ chip, and use the numerical control CNC system to process and prepare the microstructure and microchannels of the three-layer polymethacrylic acid (PMMA) chip or Preparation of PDMS replication reverse template. Add a layer of photoresist microporous film in the cell culture area, the diameter of the micropore is 5μm ~ 20μm, and then align the two layers of chips, pressurized, or thermally cross-linked to synthesize the chip to make micropores Membrane two-layer cell culture organ chip.

Embodiment 3

[0035] Example 3 Parallel implantation and co-culture of cells

[0036] Close the drug inlet and outlet, inject different cells into the upper and lower cell solution inlet and outlet, control the flow rate of the syringe pump and the liquid difference height between the inlet and outlet, thereby controlling the laminar flow speed, so that the cell solution slowly flows through the microchannel into the cell culture chamber, Thus, the implantation of cells is completed, and the plate adherent culture or the three-dimensional gel matrix culture is carried out. The cell culture medium is supplied, and the cells are cultured for a period of time. After adhering or fixed growth, the medium is circulated and flowed to promote the stable growth and proliferation of cells, and maintain the vitality and function of cells.

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Abstract

The invention relates to a two-layer cell culture system organ chip and a preparation method thereof. The organ chip comprises a two-layer cell culture system, wherein each layer comprises a culture solution micro-fluid channel, a medicament micro-fluid channel, cell culture chambers and a medicament testing tank; a micro structure and micro-fluid channels are designed on the organ chip, two cells are respectively fixed on each specific cell culture chamber, and intercellular signal transmission and interaction between the cells are performed by virtue of the micro-fluid channels. The organ chip realizes the parallel implantation and co-culture of two or more cells, is simple in operation, reduces the dose of practical samples, simplifies the cell implantation process, has the characteristics of portability, economic performance, high efficiency and accuracy, and can be used for independently performing cell seeding and culture and detection of medicament toxicity or pharmacological activity. The two-layer cell culture system organ chip is a novel organ chip with miniaturization, automation and visualization, prepared by simulating the structures and functions of a human organ, and can be used for providing the effective theoretical basis for tissue and regeneration engineering, organ transplantation and medicament evaluation.

Description

technical field [0001] The invention relates to an organ chip and a preparation method thereof, in particular to an organ chip of a two-layer cell culture system and a preparation method thereof. It belongs to the field of biomedical technology. Background technique [0002] Organ-on-a-chip is a three-dimensional cell culture system based on a multi-channel fluidic chip, which is composed of multiple cell culture partitions that simulate the human tissue and organ environment. The divisions are connected by a bionic circulation system. Organ-on-a-chip also contains integrated miniature sensing and imaging devices for real-time, online detection of the microenvironment and growth status of 3D cell aggregate growth, and the interaction between tissues and organs. Its main goal is to culture cells, tissues and organs on the chip by simulating the environment of organisms, to study and control the biological behavior of cells in the process of in vitro culture, so as to realiz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/34C12M1/12
CPCB01L3/502707B01L3/502753B01L2200/10B01L2200/12B01L2300/0819B01L2300/0887G01N33/5008
Inventor 顾忠泽郑付印赵远锦邹昕周雯婷陆洁丁海波
Owner SOUTHEAST UNIV
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