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Cell stripping method

A technology of cells and cell fluids, which is applied in the field of medicine to achieve the effect of improving the stripping efficiency, simplifying the experimental process, and improving the efficiency of stripping and purification

Pending Publication Date: 2020-03-24
JIANGSU JORTEN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The stripping detection of T cells plays a pivotal role in the health monitoring of the human body; the stripping efficiency and stripping quality of the existing T cell stripping methods still need to be further improved

Method used

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  • Cell stripping method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] S1. Put 40g of nylon wool into a beaker, add 80ml of distilled water, cover the beaker with aluminum foil and boil for 15 minutes; then cool to room temperature, pour it into a funnel with filter paper, filter out the water, and put the nylon wool again In the beaker, repeat the above steps 5 times;

[0023] S2. Spread the nylon wool in a tray covered with gauze, incubator at 30℃ and dry for 30h;

[0024] S3. Take a 50ml syringe, and configure a rubber tube with a collet at the mouth of the syringe, put nylon wool into the syringe, then wrap the syringe with tin foil, and put it into a sterilization box for sterilization.

[0025] S4. Fix the syringe, fill the syringe with a cell culture solution at 35°C, let it stand for 30 minutes, then open the chuck, and release the cell culture solution, and repeat the above steps twice.

[0026] S5. Dilute the cell liquid to be stripped to 4.9×10 with the culture medium at 35°C 7 cells / ml

[0027] S6. Pour the cell liquid in S5...

Embodiment 2

[0030] S1. Put 60g of nylon wool into a beaker, add 120ml of distilled water, cover the beaker with aluminum foil and boil for 18 minutes; then cool to room temperature, pour it into a funnel with filter paper, filter out the water, and put the nylon wool again In the beaker, repeat the above steps 6 times;

[0031] S2. Spread the nylon wool in a tray covered with gauze, incubate at 35°C and dry for 35h, cover the top of the tray with aluminum foil;

[0032] S3. Take a 50ml syringe, and configure a rubber tube with a collet at the mouth of the syringe, put nylon wool into the syringe, then wrap the syringe with tin foil, and put it into a sterilization box for sterilization.

[0033] S4. Fix the syringe, fill the syringe with a cell culture solution at 38°C, let it stand for 30 minutes, then open the chuck, and let off the cell culture solution, and repeat the above steps 3 times.

[0034] S5. Dilute the cell liquid to be stripped to 5×10 with the culture medium at 38°C 7 ce...

Embodiment 3

[0038] S1. Put 80g of nylon wool into a beaker, add 160ml of distilled water, cover the beaker with aluminum foil and boil for 20min; then cool to room temperature, pour it into a funnel with filter paper, filter out the water, and put the nylon wool again In the beaker, repeat the above steps 7 times;

[0039] S2. Spread the nylon wool in a tray covered with gauze, incubator at 45℃ and dry for 40h;

[0040] S3. Take a 50ml syringe, and configure a rubber tube with a collet at the mouth of the syringe, put nylon wool into the syringe, then wrap the syringe with tin foil, and put it into a sterilization box for sterilization.

[0041] S4. Fix the syringe, fill the syringe with cell culture solution at 40°C, let it stand for 30 minutes, then open the chuck, and let off the cell culture solution, and repeat the above steps 4 times.

[0042] S5. Dilute the cell liquid to be stripped with a culture medium at 40°C to 5.1×10 7 cells / ml

[0043] S6. Pour the cell liquid in S5 into ...

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Abstract

The invention provides a cell stripping method, and specifically relates to the technical field of medicines. The cell stripping method comprises the following steps of S1, putting 40-80g of nylon hairs in a beaker, adding 80-160ml of distilled water, covering the beaker with aluminium foil, performing boiling over for 15-20 min, performing cooling to the room temperature, and performing pouring to a funnel with a filter paper to filter off the water; S2, spreading the nylon hairs in a tray laid with a gauze, placing the tray in a temperature box of 30-45 DEG C, and performing drying for 30-40h; S3, taking a 50ml syringe, configuring a rubber tube with a clamping head at a syringe opening, placing the nylon hairs into the syringe, then wrapping the syringe with tinfoil, and placing the coated syringe in a sterilizing box for sterilization; S4, stuffing the syringe with cell culture liquid of 35-40 DEG C, and performing standing for 30 minutes; S5, diluting the cell liquid to be stripped with the culture liquid of 35-40 DEG C until the concentration is 4.9 *10<7>-5.1 *10<7> cells / ml; and S6, adding the cellular liquid in the S5 into the syringe, performing incubation under 35-40 DEG C for 50-70 min, and performing centrifugation, namely T lymphocyte stripping. Through the adoption of the cell stripping method disclosed by the invention, the stripping efficiency and the stripping quality of T lymphocyte can be improved.

Description

technical field [0001] The invention belongs to the technical field of medicine, and particularly relates to a cell stripping method. Background technique [0002] Cell dissection is a commonly used experimental method in the field of biotechnology, and it plays an important role in cell research. T lymphocytes are derived from pluripotent stem cells of bone marrow (from yolk sac and liver during embryonic stage). During the human embryonic and primary stages, a part of pluripotent stem cells or pre-T cells in the bone marrow migrate into the thymus, differentiate and mature under the induction of thymic hormones, and become immunocompetent T cells. Mature T cells are distributed to the thymus-dependent region of peripheral immune organs and settle in the peripheral immune organs, and can be recycled through lymphatic vessels, peripheral blood and tissue fluid, etc., to play the functions of cellular immunity and immune regulation. The recycling of T cells is conducive to e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2509/00
Inventor 曹振芳陈礼明杨桂娟
Owner JIANGSU JORTEN PHARMA CO LTD
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