Method for producing hiPSCs derived brain-like tissue

A production method and brain tissue technology, applied in biochemical equipment and methods, tissue culture, cells modified by introducing foreign genetic material, etc., can solve the problems of easy coagulation, large consumption, and tediousness, and achieve the goal of reducing the packaging steps Effect

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

AI Technical Summary

Problems solved by technology

[0004] In the current way of inducing brain-like production, it is basically necessary to add Matrigel. The process of operating Martigel is cumbersome, and this substance is easy to solidify and consumes a lot

Method used

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  • Method for producing hiPSCs derived brain-like tissue
  • Method for producing hiPSCs derived brain-like tissue
  • Method for producing hiPSCs derived brain-like tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The present invention provides a new method for producing brain-like tissue derived from hiPSCs, the steps of which are mainly divided into two parts:

[0052] (1) Preparation of high-throughput microcolumn chips;

[0053] (2) Generate brain organoids on a chip;

[0054] The micro-column array chip is formed by the peripheral dam structure of the chip and the surrounding micro-column 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 surrounded by four microcolumn structures;

[0055] Step (1) preparation of high-t...

Embodiment 2

[0081] The present invention provides a new method for producing brain-like tissue derived from hiPSCs, the steps of which are mainly divided into two parts:

[0082] (1) Preparation of high-throughput microcolumn chips;

[0083] (2) Generate brain organoids on a chip;

[0084] 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.

[0085] Its production steps are as follows:

[0086] Step (1) preparation of high-throughput microcolumn array chip, specifically:

[0087] (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;

[0088] (2) Fabricate a PDMS polymer chip with a high-throughput array microcolumn structure: mix PDMS and initiator 184 at a volume ratio...

Embodiment 3

[0111] The present invention provides a new method for producing brain-like tissue derived from hiPSCs, the steps of which are mainly divided into two parts:

[0112] (1) Preparation of high-throughput microcolumn chips;

[0113] (2) Generate brain organoids on a chip;

[0114] 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.

[0115] Its production steps are as follows:

[0116] Step (1) preparation of high-throughput microcolumn array chip, specifically:

[0117] (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;

[0118] (2) Fabricate a PDMS polymer chip with a high-throughput array microcolumn structure: mix PDMS and initiator 184 at a volume ratio...

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Abstract

The invention relates to a method for producing hiPSCs derived brain-like tissue. The method comprises the following steps of (1) preparing a microcolumn array chip; and (2) inducing the brain-like tissue on the chip. Brain-like organs produced by the method have relevant characteristics of in vivo brain development on gene level and protein level. The method is simple, a tedious matrigel packaging step is removed, matrigel consumption is reduced, and only additional Dorsomorphine and A83-01 need to be added at the stage of forming an embryoid body. The brain-like organs induced to 30 days have expression of cortex nerve cells (TBR1 and CTIP2), expression of mature nerve cells (TUJ1), expression of front brain (PAX6 and FOXG1) and expression of afterbrain protein (PAX2 and ISL-1), and haveexpression of excitatory neuron (vGLUT1) subtypes and inhibitory neurons (GABA) subtypes.

Description

technical field [0001] The invention belongs to organ chip technology, in particular to a method for producing brain-like tissue derived from hiPSCs. Background technique [0002] Organoids derived from neural stem cells, intestinal stem cells, embryonic stem cells, and iPSCs have been effectively developed in recent years. The characteristics of organoids are: derived from stem cells or primary cells; 3D cell cluster structure; cells with multiple functions of specific tissues and organs; cells can self-assemble into specific tissues; to a certain extent, they have certain in vivo tissues function of organs. Among them, the 3D brain derived from hiPSCs in vitro basically replicates the process of early fetal brain development, and this technology has been widely used to construct disease models, including microcephaly and Zika virus infection. [0003] At present, organoid technology still has its shortcomings and needs to be improved. Since the organoid is a 3D cell mas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10
CPCC12N5/0618C12N2513/00C12N2510/00C12N2506/45C12N2501/727C12N2501/115C12N2501/15
Inventor 秦建华崔康莉王亚清
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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