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Brain-like disease treatment tissue model based on cell three-dimensional printing technology and preparation method and application thereof

A technology for 3D printing and disease treatment, applied in biochemical equipment and methods, animal cells, tissue culture, etc., can solve problems such as brain-like tissue without breakthroughs

Active Publication Date: 2019-07-26
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above patents did not break through the blood-brain barrier between the natural brain layered tissue and blood vessels to construct brain-like tissue

Method used

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  • Brain-like disease treatment tissue model based on cell three-dimensional printing technology and preparation method and application thereof
  • Brain-like disease treatment tissue model based on cell three-dimensional printing technology and preparation method and application thereof
  • Brain-like disease treatment tissue model based on cell three-dimensional printing technology and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] as per figure 1 As shown in the process, the construction method of long-term in vitro cultured brain disease treatment tissue model based on biological three-dimensional printing includes the following steps:

[0062] (1) Design of in vitro brain-like tissue structure model:

[0063] Using the principle of bionics, based on the natural brain layered structure and structural mechanical properties, simulate the cerebral cortex with 1, 2, 3, 4, 5, 6 layers of layered structure in vitro brain-like tissue structure model, and maintain the brain-like tissue The elastic modulus of the model is 4.2kPa; the total concentration of primary neurons in each layer of cortical-like structure is 1×10 7 / mL; wherein, the concentration ratio of neurons and glial cells was 1:10.

[0064] (2) Anatomical extraction of primary nerve cells:

[0065] Primary nerve cells (including neurons and glial cells) were obtained by extracting the cerebral cortex tissue of Wistar suckling mice on the...

Embodiment 2

[0077] According to the steps in Example 1 (1)-(6) of the present invention, the in vitro three-layer epilepsy disease model is constructed (primary nerve cells (including neurons and glial cells) that are specifically used are passed through Wistar milk on the first day after birth. Rat cerebral cortex tissue was extracted. Then electrophysiological stimulation was used to construct epilepsy-like diseases.), and then the drug response characteristics of the model were tested by adding drugs with different molecular polarities.

[0078] (7-1) DNQX drug detection of relatively polar drug molecules;

[0079] Such as Figure 4The constructed brain tissue was cultured in the in vitro microenvironment of 4×4MED64 microarray electrode culture dish for 31 days. Electrophysiological studies of this brain-like structure allow the detection of excitatory post-spitting potentials (EPSPs). The neuroexcitatory inhibitor AMPA receptor antagonist (DNQX) was also directly added to the cultu...

Embodiment 3

[0081] According to the steps in Example 1 (1)-(6) of the present invention, the in vitro three-layer epilepsy disease model is constructed (primary nerve cells (including neurons and glial cells) that are specifically used are passed through Wistar milk on the first day after birth. Rat cerebral cortex tissue was extracted. Then electrophysiological stimulation was used to construct epilepsy-like diseases.), and then the drug response characteristics of the model were tested by adding drugs with different molecular polarities.

[0082] (7-2) TTX drug detection of less polar drug molecules;

[0083] Such as Figure 4 The constructed brain tissue was cultured in the in vitro microenvironment of 4×4MED64 microarray electrode culture dish for 31 days. Electrophysiological studies of this brain-like structure allow the detection of excitatory post-spitting potentials (EPSPs). Tetrodotoxin (TTX) was added to the culture environment. TTX blocks sodium ion channels, completely blo...

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Abstract

The invention discloses a brain-like disease treatment tissue model based on a cell three-dimensional printing technology and a preparation method and application thereof. The preparation method comprises the steps of mixing primary nerve cells extracted from the affected part of the brain lesion with a hydrogel material and a cross-linking agent to obtain a printing biological ink; performing printing molding on the priting biological ink based on the mechanical characteristics of the layer structure and elastic modulus of the natural cerebral cortex by adopting the biological three-dimensional printing technology and then performing tissue culture to obtain the brain-like disease treatment tissue model based on the cell three-dimensional printing technology. Based on the morphological and mechanical properties of the natural brain, the in vitro brain-like model is constructed by the biological three-dimensional printing technology. An extracellular matrix suitable for nerve cell growth and survival can be constructed by controlling the elastic modulus of the extracellular matrix, and the primary cells that are not prone to survive can be adopted for printing. The model has the function of screening drugs in different biological and chemical properties and is a drug screening model with the blood-brain-barrier similar effects.

Description

technical field [0001] The invention relates to a tissue model for treating brain-like diseases based on cell three-dimensional printing technology, a preparation method and application thereof, and belongs to the field of biomanufacturing. Background technique [0002] The central nervous system contains approximately 1.5-3.3 billion neurons, each connected to thousands of other neurons through synapses, forming complex and efficient neural networks. When nerve cells, which are the processing units of nerve signals, are damaged, the central nervous system will produce irreversible disease development, including Alzheimer's disease, Huntington's disease, Parkinson's disease and epilepsy. In order to advance theoretical research on the treatment of brain diseases and accelerate the development of industrial technologies for precision treatment, disease sample tissues are usually obtained directly from the patient's lesion area. Due to the complex structure of natural brain d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079C12N5/0793C12N5/0797C12N5/09C12Q1/02
CPCC12N5/0619C12N5/0622C12N5/0623C12N5/0693C12N2503/02C12N2513/00G01N33/5058
Inventor 宋宇张婷苏晓磊孙伟
Owner TSINGHUA UNIV
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