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Method for building hiPSCs source autism-like model

An establishment method, a technique for autism, applied in general culture methods, biochemical equipment and methods, artificial cell constructs, etc., can solve the problems of long time consumption, poor controllability, complicated operation of culturing organoids, etc., to eliminate The effect of species differences

Pending Publication Date: 2020-06-09
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, using the existing traditional cell culture technology to cultivate organoids is complex, time-consuming and poorly controllable

Method used

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  • Method for building hiPSCs source autism-like model
  • Method for building hiPSCs source autism-like model
  • Method for building hiPSCs source autism-like model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] The present invention provides a method for establishing an autism-like model derived from hiPSCs, the steps of which are mainly divided into three parts:

[0058] (1) Preparation of high-throughput microcolumn chips;

[0059] (2) Generate brain organoids on a chip;

[0060] (3) VPA treatment of 3D brains at the beginning of the induction stage;

[0061] The microcolumn array chip is formed by the peripheral dam structure of the chip and the surrounding microcolumn structure, which can be used for the growth of organoids. The formed area is the brain-like formation area. Its specific structure diagram is figure 1 . figure 1 It is a top view and a cross-sectional view of a PDMS polymer chip with an array microcolumn structure formed by a high-throughput embryoid body in specific embodiment 1; wherein 1 is a microcolumn structure, 2 is a microcolumn spacing, 3 is a chip peripheral dam structure, and 4 is a microcolumn structure. There is a brain tissue formation area...

Embodiment 2

[0092] The present invention provides a method for establishing an autism-like model derived from hiPSCs, the steps of which are mainly divided into three parts:

[0093] (2) Preparation of high-throughput microcolumn chips;

[0094] (2) Generate brain organoids on a chip;

[0095] (3) VPA treatment of 3D brains at the beginning of the induction stage;

[0096] The microcolumn array chip is formed by the peripheral dam structure of the chip and the surrounding microcolumn structure, which can be used for the growth of organoids. The formed area is the brain-like formation area.

[0097]Step (1) preparation of high-throughput microcolumn array chip, specifically:

[0098] (1) Use soft etching technology to prepare SU-8 template, which is a high-throughput concave pit, and then modify the template with low adhesion to ensure easy separation of SU-8 template and PDMS polymer;

[0099] (2) Fabricate a PDMS polymer chip with a high-throughput array microcolumn structure: mix PD...

Embodiment 3

[0126] The present invention provides a method for establishing an autism-like model derived from hiPSCs, the steps of which are mainly divided into three parts:

[0127] (3) Preparation of high-throughput microcolumn chips;

[0128] (2) Generate brain organoids on a chip;

[0129] (3) VPA treatment of 3D brains at the beginning of the induction stage;

[0130] The microcolumn array chip is formed by the peripheral dam structure of the chip and the surrounding microcolumn structure, which can be used for the growth of organoids. The formed area is the brain-like formation area.

[0131] Its production steps are as follows:

[0132] Step (1) preparation of high-throughput microcolumn array chip, specifically:

[0133] (1) Use soft etching technology to prepare SU-8 template, which is a high-throughput concave pit, and then modify the template with low adhesion to ensure easy separation of SU-8 template and PDMS polymer;

[0134] (2) Fabricate a PDMS polymer chip with a hig...

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Abstract

The invention relates to a method for building an hiPSCs source autism-like model. The method mainly comprises the steps of (1) preparing a micro-column array chip; (2) inducing a brain organoid on the chip; and (3) performing VPA treatment on a 3D brain organoid in the early period of the induction stage. According to the method, stem cells and tissue engineering are mainly combined to generate 3D brain organoid tissues; and VPA (sodium valproate) conforming to the physiological concentration is applied to simulate the influence on fetal brain development after VPA is taken by a pregnant woman with bidirectional affective disorder or epilepsy, so that a pathological mechanism of occurrence of autism is explored. The model is a novel autism in vitro model; the morphological changes of theVPA on the fetal brain development can be observed; and meanwhile, the pathogenic mechanism of the VPA increasing the fetal autism risk is explored by various measures of molecular biology and the like. Therefore, the species difference caused by animal experiments, the simplicity of cell experiments and the defects of 2D conditions can be eliminated, and a novel model and thought are provided forexploring the occurrence of autism.

Description

technical field [0001] The invention belongs to organ chip technology, and in particular relates to a method for establishing an autism-like model derived from hiPSCs. Background technique [0002] Autism is a subtype of pervasive developmental disorder, which begins in infancy and early childhood, and is mainly manifested in varying degrees of language development impairment, interpersonal communication impairment, narrow interests, and stereotyped behavior. Taking VPA during pregnancy increases the risk of autism in the fetus. Most current research on autism focuses on animal models or two-dimensional cell in vitro models. Due to the species differences between mice and humans, and the simplicity of 2D culture, a 3D human model is urgently needed. Organoids derived from neural stem cells, intestinal stem cells, embryonic stem cells, and iPSCs have been effectively developed in recent years. Organoids are characterized by their ability to accurately represent organ histo...

Claims

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Application Information

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IPC IPC(8): C12N5/00C12N5/071C12N5/079
CPCC12N5/0075C12N5/0618C12N5/0697C12N2506/45C12N2513/00
Inventor 秦建华崔康莉王亚清
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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