Cell sorting system based on endonuclease specific recognition

Abstract

The invention relates to a cell sorting system based on endonuclease specific recognition. The cell sorting system based on the endonuclease specific recognition comprises the following components: a first single-stranded nucleotide, a second single-stranded nucleotide, an antibody, a cell sorting medium and a restriction endonuclease, wherein the first single-stranded nucleotide and the second single-stranded nucleotide are complementary, and a complementary region contains a recognition site of the restriction endonuclease; the first single-stranded nucleotide, the second single-stranded nucleotide, the antibody and the cell sorting medium are all modified, the modified first single-stranded nucleotide can be connected with the modified antibody, and the modified second single-stranded nucleotide can be connected with the modified cell sorting medium. The invention also provides a cell sorting method applying the cell sorting system based on the endonuclease specific recognition, and a kit and method for sorting DC (dendritic) cells, NK (natural killer) cells and CIK (cytokine-induced killer) cells from cell samples. By adopting the cell sorting technology provided by the invention, multiple cells can be sorted at the same time, sorting of a large number of delicate cells can be carried out, and the cell yield and survival rate are high.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a cell sorting system based on endonuclease specific recognition. Background technique [0002] There are two main types of cell sorting techniques: [0003] ① Flow cytometry technology [0004] This is achieved by isolating droplets containing single cells. The cells to be tested are combined with monoclonal antibodies and fluorescent dyes to make suspension samples, and the samples enter the flow chamber under a certain gas pressure, and are arranged in a single row under the cell-free buffer package. Under the high-frequency oscillation of the ultra-high-frequency piezoelectric crystal in the flow chamber, the liquid flow is broken into uniform droplets, and the cells to be tested are included in the droplets. These droplets are charged with positive or negative charges. When the charged droplets pass through the electric field, they are deflected under the action of th...

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Problems solved by technology

[0007] The above technologies have the following disadvantages: Although the flow cytometry separation technology can sort the target cells, ① the cells must be able to be labeled by fluorescent molecules; ② can only detect single cells in suspension; ③ the sorting process is not gentle enough, causing great damage to the cells; ④It may contaminate the cells; ⑤It is not conducive to the sorting of large volume samples

Although immunomagnetic bead sorting technology can obtain target cells through positive and negative screening, ① cannot complete "one-time combination" to separate multiple cells; ② If multiple target cells need to be screened from a sample, multiple combinations and Elution, the obtained cells have poor activity and low yield; ③ multiple sorting, high cost; ④ target cells and magnetic beads cannot be completely separated, which is not conducive to follow-up research

[0008] At present, there are no reports on the technology that does little damage to cells, can sort a variety of cells at the same time, can completely separate the obtained target cells from the carrier, and is suitable for sorting a large number of fragile cells.

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