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Construction method of mature stable organoid

A construction method and organoid technology, applied in the direction of artificial cell constructs, biochemical equipment and methods, instruments, etc., can solve problems such as difficulty in ensuring reliability, lack of maturity and stability of organoids, etc.

Pending Publication Date: 2022-04-29
广州精科生物技术有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

If oligocellular structures that have not yet formed organoids or organoids that are in a state of degeneration and aging are used as in vitro experimental models, it will be difficult to guarantee the reliability of the above medical research and testing results
However, to date, there are no reports on ensuring that the cultured organoids are mature and stable

Method used

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  • Construction method of mature stable organoid

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Embodiment Construction

[0014] A method for constructing mature and stable organoids of the present invention is carried out sequentially as follows:

[0015] Step 1: Same as the prior art, take the cell suspension made from pluripotent stem cells, animal tissues and patient specimens, inoculate the stem cells or the cells enriched by centrifugation on matrix gel (such as Matrigel @ ) in vitro three-dimensional organoid culture, the cells of the embodiment of the present invention are cells obtained from bladder cancer patients to make cell suspension and enriched;

[0016] Step 2: Observe the appearance of the organoids formed in the culture system daily and measure the maximum diameter of the organoids. When the appearance standard organoids account for 50% of the total number of organoids, proceed to step 3; the appearance standard organoids are three-dimensional layered structure color Organoids with sharp, complete internal cell morphology and clear boundaries between cells, bright halos on the ...

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Abstract

The invention discloses a method for constructing a mature and stable organoid, which is used for dynamically monitoring the organoid in a three-dimensional culture process. When the appearance standard organoid (the three-dimensional multi-layer structure is bright in color and luster, the internal cellular morphology is complete, the boundary between cells is clear, the outer edge presents bright halo or the boundary is clear and complete, and the maximum diameter is 80-150m) accounts for 50% of the total number of the organoid. The mature and stable organoid is constructed by detecting the number of positive marker cell nuclei, the ratio of dead cells and apoptotic cells to the total cell number, the hypoxia state and autophagy state in the organoid and the like, the organoid is used for biomedical research and drug research and development, and the reliability of experimental results based on the organoid is improved.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, in particular to a method for constructing mature and stable organoids. Background technique [0002] Organoid (Organoid) is a multicellular three-dimensional (3D) three-dimensional structure formed by normal cells or tumor cells with proliferation ability in a semi-solid matrix gel, because the phenotype and function of the original cells are well preserved. Therefore, it can be used as a new in vitro experimental model in the fields of biomedical science research and drug development, such as tissue biology, injury repair, improvement of organ transplantation technology, disease modeling and toxicology, etc. Research; unnecessary loss of experimental animals can also be significantly reduced through "substitute test drugs". At present, most of the construction methods of organoids are cell suspensions made from pluripotent stem cells, animal tissues and patient specimens, and seeding st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/071C12Q1/02G01N33/569
CPCC12N5/0693C12N5/0685C12N5/0062G01N33/5005G01N33/56966C12N2513/00
Inventor 李宏李胜刘佳罗兴诚
Owner 广州精科生物技术有限公司
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