Cultural method for enhancing killing activity and proliferative activity of LAK (lymphokine-activated killer) cells and application

A technology of proliferation activity and killing activity, applied in the field of cell culture, can solve the problems of low proliferation activity and killing activity of LAK cells, and achieve the effect of enhancing the killing activity, proliferation activity and killing activity of LAK cells.

Active Publication Date: 2015-10-28
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of this, it is necessary to address the above-mentioned problems of low proliferation and killing activity of LAK cells and provide a culture method and application for enhancing the killing and proliferation activities of LAK cells

Method used

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  • Cultural method for enhancing killing activity and proliferative activity of LAK (lymphokine-activated killer) cells and application
  • Cultural method for enhancing killing activity and proliferative activity of LAK (lymphokine-activated killer) cells and application
  • Cultural method for enhancing killing activity and proliferative activity of LAK (lymphokine-activated killer) cells and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, the culture method of LAK cell

[0039] A culture method for LAK cells, comprising the following steps:

[0040] S1. Sorting of cells

[0041] CD14 was isolated from PBMC (peripheral blood mononuclear cell) suspension using anti-CD14 antibody magnetic beads + cells and CD14 - cell.

[0042] The preparation method of PBMC in this embodiment is as follows: after 20mL peripheral blood is centrifuged at 400-600g for 10-15min, the upper plasma is collected; ) in a sepmate centrifuge tube (purchased from STEM CELL Company), centrifuged at 600-800g for 10-20min; after centrifugation, pour out the supernatant liquid containing mononuclear cells from the sepmate centrifuge tube, and then add a certain amount of RPMI 1640 medium weight Suspend the cells and centrifuge at 400-500g for 3-5min; obtain PBMC.

[0043]In this example, the STEM CELL magnetic bead sorting kit was used to sort CD14 + monocytes, the CD14 antibody used was Easy Sep TM Human Monocyte Enr...

Embodiment 2

[0058] Embodiment 2, the culture method of LAK cell

[0059] A culture method for LAK cells, comprising the following steps:

[0060] S1. The cell sorting method is the same as that in Example 1.

[0061] Induction of S2 and DC cells

[0062] CD14 obtained by sorting step S1 + cells according to 1×10 6 The cell density per mL was resuspended with X-VIVO 15 serum-free medium containing 1000U / mL GM-CSF and 1000U / mL IL-4, then the cells were transferred to a T75 culture flask, and the cell culture flask was placed at 37°C , 5% carbon dioxide incubator for culture; observe the morphology of the cells under the microscope every day, count the cells every two days, and count the cells according to 1×10 6 A certain amount of X-VIVO 15 serum-free medium was added to the cell density per mL, and GM-CSF and IL-4 were added to make the concentration of GM-CSF 1000U / mL and the concentration of IL-4 1000U / mL ; On the 6th day, 1000 U / mL TNF-α was added to promote the maturation of DC c...

Embodiment 3

[0067] Embodiment 3, the culture method of LAK cell

[0068] A culture method for LAK cells, comprising the following steps:

[0069] S1. The cell sorting method is the same as that in Example 1.

[0070] Induction of S2 and DC cells

[0071] CD14 obtained by sorting step S1 + cells according to 1×10 6 The cell density per mL was resuspended with X-VIVO 15 serum-free medium containing 100 U / mL GM-CSF and 500 U / mL IL-4, then the cells were transferred to a T75 culture flask, and the cell culture flask was placed at 37°C , 5% carbon dioxide incubator for cultivation. Observe the morphology of the cells under the microscope every day, count the cells every two days, and count the cells according to 1×10 6 A certain amount of X-VIVO 15 serum-free medium was added to the cell density of cells / mL, and GM-CSF and IL-4 were added, so that the concentration of GM-CSF was 100U / mL and the concentration of IL-4 was 500U / mL . On the 6th day, 100 U / mL TNF-α was added to promote the m...

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Abstract

The invention relates to a cultural method for enhancing killing activity and proliferative activity of LAK (lymphokine-activated killer) cells and an application. The cultural method for enhancing the killing activity and proliferative activity of the LAK cells is co-culturing the LAK cells and DC (dendritic cells). The application of the DC cells is to enhance the killing activity and proliferative activity of the LAK cells. According to the cultural method, through co-culturing the LAK cells and the DC cells, the proliferation of the LAK cells is promoted, and the killing activity of the LAK cells is enhanced.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a culture method and application for enhancing LAK cell killing activity and proliferation activity. Background technique [0002] With the aging population, the destruction of the ecological environment, the emergence of unhealthy lifestyles and food safety issues, the incidence of cancer in my country has continued to rise for many years, and it has become a public health problem and even a social problem that must be attached great importance to. China urgently needs to declare war on cancer. According to the World Cancer Report, in 2012, the number of cancer cases in China was 3.065 million, accounting for about one-fifth of the global incidence; the number of cancer deaths was 2.205 million, accounting for about one-quarter of the global cancer deaths. At present, the incidence and mortality of cancer in my country are showing a continuous growth trend. The incidence of cancer in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783C12N5/0784
Inventor 陈海佳王一飞葛啸虎罗二梅
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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