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Method for culturing cells B for long time

A B cell and cell technology, applied in the field of cell culture, can solve the problems of limited B cell proliferation multiple, cannot meet clinical use, and difficult to source, and achieve the effect of enhancing phagocytosis and killing effect, reducing risk, and multiplying multiple multiples.

Inactive Publication Date: 2016-11-23
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The above-mentioned technology is still quite limited in terms of the multiplier of B cells, which cannot meet the needs of clinical use at all; and the existing technology requires the use of human serum, which is difficult to source and also has a high risk of infectious diseases, which is also prohibited in clinical practice.

Method used

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  • Method for culturing cells B for long time
  • Method for culturing cells B for long time
  • Method for culturing cells B for long time

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Medium formula:

[0036]

[0037]

[0038] (1) Collect 20ml of peripheral blood from healthy people;

[0039] (2) Collect plasma after centrifugation at 500g;

[0040] (3) Dilute the lower blood cells 1:1 with physiological saline, then add the diluted solution to the sepmate centrifuge tube (purchased from STEM CELL company) that has Ficoll separation solution (purchased from STEM CELL company), and centrifuge for 10-20 min;

[0041] (4) Pour the upper liquid from the centrifuge tube after centrifugation, and add RPMI 1640 medium to resuspend the cells. The amount of medium is determined by the number of cells, and the cell density is greater than 1×10. 7 / ml, centrifugal counting detection flow data;

[0042] (5) Take 1×10 among them 7 Cells, according to 1×10 6 The cell density of cells / ml is added to the culture medium of the present invention at 37°C, 5% CO2 for culture, and the fluid is supplemented every 3-4 days to ensure that the cell density is 1×10 6 Pcs / ml.

[0043] (6...

Embodiment 2

[0046] Medium formula:

[0047]

[0048]

[0049] (1) Collect 20ml of peripheral blood from healthy people;

[0050] (2) Collect plasma after centrifugation at 500g;

[0051] (3) Dilute the lower layer blood cells 1:1 with physiological saline, and then add the diluent to the sepmate centrifuge tube (purchased from STEM CELL company) that has Ficoll separation solution (purchased from STEM CELL company), and centrifuge for 10-20 min;

[0052] (4) Pour the upper liquid from the centrifuge tube after centrifugation, and add RPMI 1640 medium to resuspend the cells. The amount of medium is determined by the number of cells, and the cell density is greater than 1×10. 7 / ml, centrifugal counting detection flow data;

[0053] (5) Take 1×10 among them 7 Cells, according to 1×10 6 Cell density per ml added to the culture medium of the present invention at 37℃, 5% CO 2 Incubate and replenish the fluid every 3-4 days to ensure that the cell density is 1×10 6 Pcs / ml.

[0054] (6) On the 21st day of ...

Embodiment 3

[0057] Medium formula:

[0058]

[0059]

[0060] (1) Collect 20ml of peripheral blood from healthy people;

[0061] (2) Collect plasma after centrifugation at 500g;

[0062] (3) Dilute the lower layer blood cells 1:1 with physiological saline, and then add the diluent to the sepmate centrifuge tube (purchased from STEM CELL company) that has Ficoll separation solution (purchased from STEM CELL company), and centrifuge for 10-20 min;

[0063] (4) Pour the upper liquid out of the centrifuge tube after centrifugation, and add RPMI 1640 medium to resuspend the cells. The amount of medium is determined by the number of cells, and the cell density is greater than 1×10. 7 / ml, centrifugal counting detection flow data;

[0064] (5) Take 1×10 among them 7 Cells, according to 1×10 6 The cell density of cells / ml is added to the culture medium of the present invention at 37°C, 5% CO2 for culture, and the fluid is supplemented every 3-4 days to ensure that the cell density is 1×10 6 Pcs / ml.

[0065]...

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Abstract

The invention relates to the field of cell culture, in particular to a method for culturing cells B for a long time. According to the method, firstly, the proliferation number of the cells B is increased; then, the culture by using human serum is avoided; the cell safety is greatly enhanced.

Description

Technical field [0001] The invention relates to the field of cell culture, in particular to a method for culturing B cells for a long time. Background technique [0002] B lymphocytes are a very important immune cell, and T lymphocytes together constitute the human immune system. In the immune response, B lymphocytes play a very powerful role. First of all, in humoral immunity, the main function is to become plasma cells that secrete immunoglobulins to produce antibodies, or to become memory B cells; at the same time, B cells are also very important for antigen extraction. Presenting cells can engulf, recognize, and present antigens to T cells, thereby guiding the killing effect of T cells. Activated B cells can continuously express major histocompatibility complex (MHC) class I and class II molecules, effectively present antigens, and can also express CD80, which is an indispensable part of the human immune system. [0003] At present, B cells are cultured in IMDM culture medium...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0781
CPCC12N5/0635C12N2500/25C12N2500/84C12N2501/22C12N2501/2304C12N2501/2316C12N2501/26C12N2501/33
Inventor 陈海佳王一飞葛啸虎万桦张维敏
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD