Liver cancer stem cell vaccine as well as preparation method and application thereof

A technology of liver cancer stem cells and liver cancer cells, which is applied in the field of tumor stem cell vaccines and their preparation and application, can solve the problems of low curative effect and lack of liver cancer stem cell vaccines, and achieve the effects of enhancing curative effect, improving the immune function of the body, and enhancing the ability to absorb antigens

Active Publication Date: 2015-01-07
广州复大医疗有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, radiotherapy and chemotherapy have little effect on the small number of stem cell-like cells present in the tumor, and the proliferation of the remaining liver cancer stem cells after treatment is enough to promote one of liver cancer recurrence and metastasis.
At present, there is still a lack of effective liver cancer stem cell vaccines, and those skilled in the art urgently need to make a breakthrough in this regard

Method used

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  • Liver cancer stem cell vaccine as well as preparation method and application thereof
  • Liver cancer stem cell vaccine as well as preparation method and application thereof
  • Liver cancer stem cell vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: Preparation of Liver Cancer Stem Cell Vaccine

[0042] In this example, liver cancer stem cell vaccines were prepared using liver cancer cells stored in liquid nitrogen as raw materials.

[0043] 1. Put the liver cancer cell QGY-7701 stored in liquid nitrogen or -80℃ in a 37℃ water bath to dissolve quickly;

[0044] 2. Mix the thawed tumor cell suspension with 8.0ml medium (RPMI1640+10% FBS), and centrifuge at 1500 rpm for 8 minutes;

[0045] 3. Discard the supernatant, then absorb 8.0ml culture medium to mix the cell pellet, then centrifuge at 1500 rpm for 8 minutes, discard the supernatant, mix the cell pellet with 1.0ml culture medium, and set aside;

[0046] 4. Take another 75cm 2 Square bottle, add 14.0ml of culture medium, add the suspension containing tumor cells (1.0ml) prepared above into this square bottle, at 37°C, 5% CO 2 cultured in an incubator;

[0047] 5. Tumor cells grow on the wall. After 3-4 days, when the tumor cells grow to 80%-95% mo...

Embodiment 2

[0064] Example 2: Curative Effect Observation After Injection of Liver Cancer Stem Cell Vaccine

[0065] The liver cancer stem cell vaccine prepared in Example 1 was applied to the treatment of liver cancer patients. The detailed experimental process was as follows: 30 liver cancer patients were randomly selected and divided into two groups, 15 of which received the vaccine injection treatment prepared in Example 1 as a treatment. group; the other 15 people who did not receive vaccine treatment were used as the control group. Before injecting patients, 5ml of blood was drawn from each patient, and the number of peripheral blood lymphocytes (T BNK detection kit) and lymphocyte function (6colour T BNK With Trucount detection kit) were detected, and the report sheet was obtained through flow cytometry detection. For patients in the treatment group, 500ul of the vaccine dose was subcutaneously injected into the liver cancer patients, and then injected again after one week interval...

Embodiment 3

[0070] Example 3: Humoral immune response induced by the tumor stem cell vaccine prepared in Example 1

[0071] 30 routine patients in the embodiment 1 are equally divided into two groups (no vaccine injection group-control group, vaccination group-experimental group), carry out according to the following method respectively:

[0072] Collect 5ml of the whole blood of the patient on the 14th day, and separate the plasma with the lymphocyte separation medium. The details are as follows: Take 5ml of fresh anticoagulant blood, mix it with normal saline for injection at a ratio of 1:1, and carefully add it to 2ml of the lymphocyte separation medium , centrifuge at 400g (horizontal rotor with a radius of 15cm) for 20 minutes. At this time, the centrifuge tube is divided into four layers, and the uppermost layer is plasma. The IgG content in plasma was detected by ELISA.

[0073] The result is as image 3 As shown, compared with the control group, P image 3 It can be seen that aft...

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Abstract

The invention discloses a cancer stem cell vaccine as well as a preparation method and application thereof. The liver cancer stem cell vaccine comprises inactivated liver cancer stem cells, mannatide and fat milk, which are mixed in a volume ratio of 3 to 1 to 1. The preparation method of the liver cancer stem cell vaccine comprises the following steps: obtaining the liver cancer stem cells; inactivating the liver cancer stem cells; freeze-thawing the inactivated liver cancer stem cells in normal saline and uniformly mixing the liver cancer stem cells with the adjuvants, namely mannatide and fat milk, thus obtaining the liver cancer stem cell vaccine. The invention provides the application of the liver cancer stem cell vaccine obtained by the preparation method to preparation of drugs for treating liver cancers. The liver cancer stem cell vaccine provides a new approach for treatment of liver cancers and preparation of the vaccines of liver cancers, has positive treatment effects on patients with liver cancers, can enhance the immune functions of the patients, and plays a positive role in lengthening the survival time of the patients.

Description

technical field [0001] The invention relates to biomedical engineering technology, and relates to a tumor stem cell vaccine and its preparation and application. Background technique [0002] Malignant tumor is a systemic chronic disease. In patients with middle and advanced stages, there are not only obvious mass tumors in the organs during imaging examinations, but also a large number of invisible circulating tumor cells and cancer stem cells (Cancer stem cells, CSCs) in the peripheral blood, ready to settle in the body at any time Susceptible sites develop into new metastases, which is why the tumor cannot be cured. Traditional tumor treatment includes 4 main therapies, namely surgery, chemotherapy, radiotherapy and immunotherapy. Usually, after 3-5 years of treatment with the first three therapies, the tumor will metastasize to peripheral organs or distant organs. With the in-depth research on CSC, it has been clear that the reason for recurrence and metastasis is not ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/39A61K47/42C12N5/095A61P35/00
Inventor 陈继冰林茂左建生刘建国徐克成牛立志
Owner 广州复大医疗有限公司
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