Single cell separator, application thereof in single cell separation process and monoclonal cell preparation method

A single-cell separation and single-cell technology, applied in the field of single-cell separation, can solve the problems of complicated operation of limited dilution method and high requirements for flow cytometer method equipment, save experimental time, avoid cell counting and repeated dilution, and improve preparation. The effect of efficiency

Pending Publication Date: 2021-02-23
天康生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The first object of the present invention is to provide a single cell separator, which can quickly separate and extract single cells from the cell fluid, and effectively alleviate the complicated operation of the limited dilution method in the existing single cell separation method And the flow cytometry method has high requirements for equipment

Method used

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  • Single cell separator, application thereof in single cell separation process and monoclonal cell preparation method
  • Single cell separator, application thereof in single cell separation process and monoclonal cell preparation method
  • Single cell separator, application thereof in single cell separation process and monoclonal cell preparation method

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preparation example Construction

[0046] According to one aspect of the present invention, a method for preparing monoclonal cells, the method comprises the following steps:

[0047] Provide cell liquid containing suspended cells, then use the above-mentioned single cell separator to absorb single cells in the cell liquid under a microscope, and then culture the single cells to obtain monoclonal cells;

[0048] Or, provide cell liquid containing adherent cells, then use the above-mentioned single cell separator to absorb single cells in the cell liquid under a microscope, and then culture the single cells to obtain monoclonal cells.

[0049] The method for preparing monoclonal cells provided by the present invention, the method firstly provides cell liquid containing adherent cells or suspension cells, then under a microscope, uses the above-mentioned single cell separator to absorb the single cells in the cell liquid, and then the single cells Cultured to obtain monoclonal cells. The above method realizes th...

Embodiment 1

[0057] figure 1 Schematic diagram of the structure of the single cell separator provided in Example 1; wherein: liquid suction device 1, that is, a 20ul pipette; pipette needle 2, that is, a syringe needle that is folded at about 90° at a distance of about 0.8 cm from the needle mouth.

[0058] like figure 1 Shown, a kind of single-cell separator, the preparation method of described single-cell separator, comprises the following steps:

[0059] First, take a syringe needle, cut off the tip of the needle with scissors, and grind it flat; then use a hemostat to fold the needle at a distance of about 0.8 cm from the needle mouth to about 90°; then, fold the needle at about 90° The tail of the syringe needle was connected to a 20ul pipette to prepare a single cell isolation device.

Embodiment 2

[0061] A kind of single cell separator, the preparation method of described single cell separator, comprises the following steps:

[0062] First, take a syringe needle, cut off the tip of the needle with scissors, and grind it flat; then use a hemostatic forceps to fold the needle at about 0.5cm from the needle mouth to about 75°; then, fold the needle at about 75° The tail of the syringe needle was connected to a 20ul pipette to prepare a single cell isolation device.

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Abstract

The invention provides a single cell separator, application thereof in a single cell separation process and a monoclonal cell preparation method, and relates to the technical field of single cell separation. The single cell separator comprises a liquid transfer needle and a liquid suction device, wherein the liquid transfer needle tightly communicates with the liquid suction device without leakage; and the liquid transfer needle comprises an intravenous injection needle which is bent at an angle of 75 to 90 degrees at a position 0.5 to 1 cm away from a needle opening. By using the single cellseparator provided by the invention, single cells can be quickly separated and extracted from cell sap, and the problems of complicated operation of a limited dilution method and high equipment requirement of a flow cytometer method in an existing single cell separation method are effectively relieved.

Description

technical field [0001] The invention relates to the technical field of single cell separation, in particular to a single cell separator and its application in the process of separating single cells and a method for preparing monoclonal cells. Background technique [0002] In population cells, due to the heterogeneity between cells, even the same cell line of different generations or the sensitivity of cells between different cell lines of the same generation to viruses, the integrity of virion packaging or virus preparation effectiveness will vary. Therefore, single cell isolation is of great significance to overcome cell heterogeneity. In addition, for experiments such as cell transfection and infection, it is very necessary to select a single cell to establish a clone of a specific gene or express a specific protein. Because only a single gene cell population can maintain population stability to the greatest extent. In order to screen high-yield and high-quality cells f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/26C12N5/00
CPCC12M33/04C12N5/00C12N2509/10
Inventor 贺笋张迪赵毅张丹刘宏
Owner 天康生物制药有限公司
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