Preparation method of tumor organoid

A technology of organoids and tumors, which is applied in the field of tumor organoid preparation, can solve the problems of difficult detection of tumor organoids, and achieve the effects of short culture time, good connection, and not easy to scatter

Pending Publication Date: 2021-04-02
GUANGDONG PROCAPZOOM BIOSCIENCES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, there are certain difficu...

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  • Preparation method of tumor organoid
  • Preparation method of tumor organoid
  • Preparation method of tumor organoid

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Example 1: Preparation of tumor organoids

[0038] 1) Obtain a biopsy from a cancer or tumor patient.

[0039] 2) The biopsy tissue is soaked in tissue preservation solution to ensure tissue activity.

[0040] 3) Spread a layer of methacrylylated gelatin on a cell culture plate (96-well plate, 48-well plate, 24-well plate, 12-well plate, or 6-well plate), irradiate blue light to solidify the gel, and then spread A layer of pigskin gelatin is applied to adhere to the surface of the methacrylated gelatin.

[0041] 4) Take the tissue out of the preservation solution, place it in a cell culture dish, add cell culture medium, chop the tissue in the cell culture medium, and the volume of small clumps is below 0.5mm×0.5mm.

[0042] 5) Mix the minced tissue-medium suspension and pass it through a sterile filter with a pore size of 100 um.

[0043] 6) The liquid passing through the filter is either discarded or used to cultivate primary cells, and the tissue mass intercepted ...

Embodiment 2

[0055] Embodiment 2: drug screening

[0056] For the testing of some new drugs, commonly used models in laboratories are PDX models (Patient-Derived tumor Xenograft) or CDX models (Cell-line-Derived tumor Xenograft). The PDX model is to extract the patient's primary cells and transplant them under the skin of nude mice or other immune-deficient mice to make the cells grow into tumors. After that, the mice were injected with the drug, and after several cycles of injection, the mass and volume of the tumor were measured to see if it was reduced compared to the control group. The problem with this is that although the experimental data count as in vivo experimental data, the growth environment in mice is very different from the growth environment in humans. For example, normal human neural stem cells cannot grow when transplanted into mice. Therefore, it is unknown whether the characteristics of patient tumor cells that can grow smoothly in mice have changed.

[0057] The grow...

Embodiment 3

[0059] Example 3: Drug Testing

[0060] With the patient's informed consent, the Shantou Cancer Hospital obtained the patient's recurrent cervical cancer tissue sample, put the tissue sample into the tissue preservation solution, and then obtained the tumor organoid according to the index method in Example 1. This tumor organoid was used for dosing experiments. The tested drugs included gemcitabine, paclitaxel, gemcitabine-romidepsin-cisplatin combination, C188-9, ivermectin-cisplatin combination, and BBI608-paclitaxel combination. Among them, C188-9 and BBI608 are new small molecular drugs, and the others are chemotherapy drugs commonly used in hospitals. Test results such as Figure 11 As shown, it can be seen from the figure that the tumor organoid is more sensitive to the combination of ivermectin-cisplatin and BBI608-paclitaxel, and the effective C188-9 in the tumor organoid test in Example 2 No obvious drug effect was shown.

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Abstract

The invention provides a preparation method of a tumor organoid. The preparation method comprises the following steps: acquiring a tumor tissue sample; spreading a layer of methacrylated gelatin on acell culture plate to solidify the gelatin and spreading a layer of pigskin gelatin or polylysine gel to solidify the gelatin; placing the sample tissue into a sterile container and chopping the sample tissue in a cell culture medium to obtain a tissue block mass; passing a chopped sample tissue-culture medium suspension through a sterile filter screen; flushing the tissue block mass intercepted by the filter screen to a new sterile container by using the cell culture medium, adding serum from the tumor tissue sample, and performing uniform mixing; adding an obtained mixture into wells of thecell culture plate and performing culture overnight; and observing the tissue block mass and obtaining a tumor organoid when the block mass is adsorbed to an upper layer of a gelatin surface in a semi-inlaying manner and does not float back and forth along with liquid. The method has short time for preparing the tumor organoid, cells in the organoid are well connected and not prone to scattering,whether the organoid has activity or not can be conveniently seen, and the method can also be used for detecting a cell-killing ability.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing tumor organoids. Background technique [0002] Organoid refers to a biologically active cell group (cluster) composed of multiple cells. Compared with single cells, it is closer to the actual situation of multiple cells growing together. The history of organoids can be roughly traced back to the early 21st century. Lancaster, a neuroscientist at the University of Cambridge, discovered that unattached brain cells can form cell clusters and are biologically active when cultivating brain cells, so they are closer to the state where cells work together or grow together. [0003] Organoid culture technology can use cells derived from tissues, embryonic stem cells and adult stem cells to induce differentiation in vitro into functional cell clusters with the basic characteristics of their source organs, providing a new in vitro model for simulating living...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/18C12Q1/02
CPCC12N5/0693G01N33/5011G01N33/5044C12N2533/54C12N2533/32C12N2503/02
Inventor 曾皓宇关天李芸林燕纯
Owner GUANGDONG PROCAPZOOM BIOSCIENCES CO LTD
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