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Preparation method of lung cancer stem cell vaccine for obviously improving immunocompetence

A technology of lung cancer stem cells and immune ability, applied in the field of vaccine preparation, can solve the problems such as the difficulty in preparing lung cancer vaccines, and achieve the effects of enhancing immunity, controlling growth, and significant curative effect

Inactive Publication Date: 2017-05-17
CHENGDU YUYA TECH
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  • Summary
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  • Claims
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Problems solved by technology

Similarly, although mannan or mannan peptides are also commonly used anti-tumor adjuvants (such as "Oligomannose-coated liposomes as a therapeutic antigen-delivery and an adjuvant vehicle for induction of in vivo tumor immunit"), it is also difficult to prepare a lung cancer vaccine

Method used

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  • Preparation method of lung cancer stem cell vaccine for obviously improving immunocompetence

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Experimental program
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Embodiment

[0023] (1) The resuscitated lung cancer stem cells were cultured in DMEM full medium containing 8% FBS, the culture temperature was 37 °C, the humidity was 85%, CO 2 The concentration is 0.5%, cultured for 3-4 days, and then the cultured cells were divided into 1:3 plates, and then cultured with the same medium under the same culture conditions for 3 days, the cells showed spindles or similar spindles ;

[0024] (2) Utilize the result of step (1) 60 Co was irradiated with a dose of 3Gy;

[0025] (3) Select CD133-positive stem cells, freeze and thaw them several times after culture to inactivate them, and then resuspend the stem cells in normal saline to make the cell concentration 5×10 5 a / ml;

[0026] (4) According to the ratio of volume ratio 4:1.5:2, the result of step (3), chitosan saline solution and laminarin saline solution were mixed, after mixing, the concentration of chitosan was 2%, and the concentration of laminarin was 2%. The concentration is 2.5%.

experiment example

[0030] Ten Kunming rats implanted with lung cancer tumors were injected with the vaccine obtained in the examples as experimental group 1, another 10 Kunming rats were injected with the product of the comparative example as experimental group 2, and another 10 Kunming rats were injected with normal saline as the control group. The injection volume of both the experimental group and the control group was 200 ul, and the number of peripheral lymphocytes, lymphocyte function and tumor volume were detected 15 days later. The results are shown in Table 1, Table 2 and Table 3.

[0031] Table 1

[0032]

[0033] Note: *P<0.01

[0034] Experimental group 1 Experimental group 2 control group IFN-γ (pg / ml) 1.69±0.11 0.81±0.13 0.74±0.20 TNF (pg / ml) 1.57±0.26 0.76±0.15 0.69±0.12 IL-4 (pg / ml) 2.95±0.11 2.01±0.15 1.88±0.24

[0035] Note: *P<0.01

[0036] Experimental group 1 Experimental group 2 control group Tumor volume ...

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Abstract

The invention provides a preparation method of a lung cancer stem cell vaccine for obviously improving immunocompetence. The method comprises the following steps of (1) culturing the recovered lung cancer stem cells in a DMEM whole culture medium, wherein the DMEM whole culture medium comprises 8 percent of FBS; the culture temperature is 37 DEG C; the humidity is 85 percent; the CO2 concentration is 0.5 percent; the culture time is 3 to 4 days; then, performing 1:3 dish separation on the cultured cell; next, performing culture for 3 days by using the same culture medium under the same culture conditions so that the cells show spindles or are similar to spindles; (2) performing irradiation treatment by 60Co at the dosage of 3 to 4 Gy; (3) selecting CD133 positive stem cells; performing inactivation treatment after the culture; resuspending the stem cells by normal saline so that the cell concentration is 1*10<5> to 1*10<6> / ml; (4) according to the volume ratio of 4:1.5:2, mixing the materials obtained in the step (3), chitosan normal saline solution and laminarin normal saline solution, and after the mixing, the chitosan concentration is 2 percent, and the laminarin concentration is 2.5 percent. The obtained vaccine has the obvious curative effect on the lung cancer.

Description

technical field [0001] The invention belongs to the field of vaccine preparation, and in particular relates to a preparation method of a lung cancer stem cell vaccine that significantly improves immunity. Background technique [0002] Tumor vaccines use tumor cells or tumor antigens to induce the body's specific cellular and humoral immune responses to regulate the body's immune function and achieve the purpose of treating tumors. Therapeutic tumor vaccines can induce systemic specific antitumor effects through active immunization. According to different preparation methods, vaccines are mainly divided into: cellular vaccines, polypeptide vaccines and protein vaccines, nucleic acid vaccines, and anti-idiotypic antibody vaccines. [0003] Cancer stem cell vaccine is a recently emerging research method, and corresponding research progress has been made in liver cancer and ovarian cancer. Since the immunogenicity of tumor stem cells is very weak, one of the main tasks of deve...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00C12N5/095
CPCA61K39/0011A61K2039/5152C12N5/0695C12N2529/00
Inventor 向红先
Owner CHENGDU YUYA TECH
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