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Thylakoid membrane and NK cell fused cell and construction method therefor

A construction method and technology of NK cells, applied in the field of fusion cells of thylakoid membrane and NK cells and their construction, can solve problems such as influence, and achieve the effects of low acquisition cost, shortened extraction time, and effective fusion

Active Publication Date: 2021-07-23
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Despite their activity in controlling tumor growth, NK cells are susceptible to multiple immunosuppressive mechanisms in the tumor microenvironment (TME)

Method used

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  • Thylakoid membrane and NK cell fused cell and construction method therefor
  • Thylakoid membrane and NK cell fused cell and construction method therefor
  • Thylakoid membrane and NK cell fused cell and construction method therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0047] 1. Extraction of thylakoid membranes

[0048] Isolation of thylakoid membranes from fresh spinach.

[0049] Specifically, 100 g of spinach leaves were washed with pure water, and then kept overnight at 4°C. Homogenize the spinach leaves in a juicer, and add 300mL of homogenization buffer (20mM Tricine, 0.4M sucrose, 2mM ascorbic acid, 2mM MgCl 2 , 40 mM NaCl, 0.2% BSA, pH 7.8). Then filter the leaf fragments with ten layers of gauze and collect the filtrate. The filtrate was centrifuged at 8000 rpm for 10 min to collect complete chloroplasts.

[0050] The obtained chloroplasts were resuspended in hypotonic lysis buffer (10 mM HEPES, pH 8.0) for 2 h, centrifuged at 8000 rpm for 10 min, and the matrix extract in the supernatant was removed.

[0051] Finally, the thylakoids were suspended in water, sonicated for 5 minutes, and centrifuged at 16,000 rpm for 15 minutes to take out the extract from the tube. The obtained thylakoid membrane was a dark green precipitate.

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Abstract

The invention provides a thylakoid membrane and NK cell fused cell and a construction method therefor and relates to the technical field of biology. The construction method for the thylakoid membrane and NK cell fused cell comprises the step: fusing thylakoid nano-vesicles with NK cells, wherein the thylakoid nano-vesicles, of which the particle size is 200nm-500nm and the concentration is 5 to 80 micrograms, are added into per 4*10<6> to 6*10<6> NK cells, and the cells are subjected to induced fusion through polyethylene glycol. According to the thylakoid membrane and NK cell fused cell and the construction method therefor, the NK cells and a thylakoid membrane are fused to form an engineering fused cell for the first time, and the fused cell simultaneously has catalase activity of the thylakoid membrane and tumor targeting and killing properties of the NK cells.

Description

technical field [0001] The invention relates to the technical field of biology, in particular to a fusion cell of thylakoid membrane and NK cell and a construction method thereof. Background technique [0002] Adoptive immune cell therapy collects the body's own immune cells, cultivates them in vitro to increase their number and enhance their targeted killing function, and then infuse them back into the patient's body to kill pathogens and cancer cells in blood and tissues , Mutated cells. Adoptive immune cell therapy has proven ability to overcome the bottleneck of cancer treatment. Natural killer cells (NK cells) account for 10-20% of lymphocytes in blood, are one of the most important natural immune cells in immune monitoring, and have been found to have rapid and effective immunity to cancer cells. Unlike T cells and B cells, NK cells have the ability to directly target cancer cells through inhibitory or activating cell surface receptors, and can kill cancer cells by r...

Claims

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Application Information

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IPC IPC(8): C12N15/08C12N5/28
CPCC12N5/12C12N2500/50
Inventor 谢海燕雷垚武广昊刘厚莉
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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