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Human hemangioma animal model and high-success-ratio construction method thereof

A technology of animal model and construction method, applied in animal husbandry and other fields, can solve problems such as low tumor formation rate, ulcer, bleeding, infection, heart failure, life-threatening, etc., achieve high tumor formation rate, strong cell proliferation, and easy operation Effect

Active Publication Date: 2020-01-17
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some superficial tumors may affect the appearance and physiological function of patients, and some tumors may cause ulcers, bleeding, infection, high-flow congestive heart failure, and even life-threatening
[0003] At present, the formation mechanism of hemangioma is still unclear, and there is no simple and easy method for constructing animal models of human hemangioma. This problem limits the in-depth research on the pathogenesis and treatment of hemangioma to a certain extent.
However, this method is complex and cumbersome to operate and the tumor formation rate is not high

Method used

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  • Human hemangioma animal model and high-success-ratio construction method thereof
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  • Human hemangioma animal model and high-success-ratio construction method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] This embodiment provides a method for constructing a human hemangioma animal model, and the specific steps are as follows:

[0039] ①Cultivation of hemangioma endothelial cells (HemECs)

[0040] a. Purchasing or separating and extracting human hemangioma endothelial cells;

[0041] b. Cells were cultured and proliferated in DMEM complete medium (10% FBS + 1% double antibody + 89% DMEM basal medium), and counted by hemocytometer.

[0042] ② Hemangioma endothelial cells (HemECs) to construct hemangioma animal model mother tumor

[0043] a. Digest the cultured hemangioma endothelial cells with 0.25% trypsin;

[0044] b. After counting, take 1×10 6 Centrifuge the cell digestion solution in a 15mL centrifuge tube at 1200rpm for 5 minutes, and discard the supernatant;

[0045] c. Add PBS to resuspend the cells, centrifuge to discard the supernatant, then add PBS to resuspend the cells, centrifuge to discard the supernatant, add 100 μL PBS to resuspend the cells.

[0046]...

Embodiment 2

[0060] This embodiment provides a method for constructing a human hemangioma animal model, and the specific steps are as follows:

[0061] ①Cultivation of hemangioma stem cells

[0062] a. Purchasing or separating and extracting human hemangioma stem cells;

[0063] b. Cells were cultured and proliferated in DMEM complete medium (10% FBS + 1% double antibody + 89% DMEM basal medium), and counted by hemocytometer.

[0064] ② Hemangioma stem cells to construct hemangioma animal model mother tumor

[0065] a. Digest the cultured hemangioma stem cells with 0.25% trypsin;

[0066] b. After counting, take 1×10 6 Centrifuge the cell digestion solution in a 15mL centrifuge tube at 1200rpm for 5 minutes, and discard the supernatant;

[0067] c. Add PBS to resuspend the cells, centrifuge to discard the supernatant, then add PBS to resuspend the cells, centrifuge to discard the supernatant, add 100 μL PBS to resuspend the cells.

[0068] d. Take a 1mL syringe, draw 100μL of cell sus...

Embodiment 3

[0076] This embodiment provides a method for constructing a human hemangioma animal model, the specific steps are basically the same as those in Example 1, and the steps are as follows:

[0077] ① Culture and expansion of human hemangioma endothelial cells.

[0078] ② Put 1×10 6 Hemangioma endothelial cells were resuspended evenly in 100 μL PBS;

[0079] ③ Inoculate the above-mentioned human hemangioma endothelial cells into immunodeficient BALB / c-nu nude mice subcutaneously with a syringe;

[0080] ④ intraperitoneal injection of estradiol 0.5 mg to obtain the mother tumor;

[0081] ⑤ Remove the mother tumor, peel it off and wash it with PBS;

[0082] ⑥ Divide the mother tumor into 30mm 3 Small pieces of (tumor tissue);

[0083] ⑦The tumor tissue was inoculated subcutaneously into immunodeficient BALB / c-nu nude mice with implantation trocar.

[0084] The shape and tissue structure of the hemangioma animal model established by the present invention are very similar to hum...

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Abstract

The invention discloses a human hemangioma animal model and a high-success-ratio construction method thereof. The construction method includes the steps: S1 taking isolated human hemangioma cells forstandby application; S2 uniformly resuspending the human hemangioma cells by PBS (phosphate buffer); S3 subcutaneously inoculating the resuspended human hemangioma cells into immune deficiency animals; S4 performing intraperitoneal injection with estradiol to obtain a parent tumor; S5 picking the parent tumor, and completely peeling and flushing the parent tumor; S6 dividing the flushed parent tumor into tumor tissues of 30-70mm<3>; S7 subcutaneously inoculating the tumor tissues into the immune deficiency animals, and feeding the tumor tissues to obtain the human hemangioma animal model. Themethod has the advantages that hemangioma formation rate is high and can reach 100%, cell components of the formed hemangioma are completely humanized, and the model is applicable to research pathogenesis, new drug evaluation and treatment methods of the human hemangioma.

Description

technical field [0001] The invention relates to an animal model and a construction method thereof, in particular to a human hemangioma animal model constructed from human hemangioblastoma constructed by human hemangioma endothelial cells and a construction method thereof with a high success rate. Background technique [0002] Hemangioma is one of the most common angiogenic diseases in infants and young children. It occurs in the head, face and extremities, accounting for 2.5-12% of all newborns. Under normal circumstances, the tumor body begins to disappear one year after birth, and it can naturally and slowly disappear and return to normal within 10 years. However, some superficial tumors may affect the appearance and physiological function of patients, and some tumors may cause ulcers, bleeding, infection, high-flow congestive heart failure, and even life-threatening. [0003] At present, the formation mechanism of hemangioma is still unclear, and there is no simple and e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/02
CPCA01K67/02
Inventor 袁伟恩郭晓霜卫旻炎苑子涵徐阳陈璇
Owner SHANGHAI JIAO TONG UNIV
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