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Method for detecting in-vitro inhibition of lymphocyte proliferation of human amniotic epithelial cells

A lymphocyte proliferation and epithelial cell technology, applied in the field of lymphocyte proliferation inhibition, can solve the problems of slow reproduction, large data fluctuation, and difference in the quality of human amniotic cells.

Pending Publication Date: 2021-04-30
华夏源细胞工程集团股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, human amnion cells will enter the logarithmic growth phase after three days of culture, multiply slowly, and have a slow inhibitory effect on lymphocyte proliferation. At the same time, due to differences in cell culture proliferation and deproliferation methods, the quality of human amnion cells also varies greatly. , when the lymphocyte proliferation inhibition is detected in vitro, the data fluctuates greatly and is unstable

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  • Method for detecting in-vitro inhibition of lymphocyte proliferation of human amniotic epithelial cells
  • Method for detecting in-vitro inhibition of lymphocyte proliferation of human amniotic epithelial cells

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0061]Example 1 of the present invention provides a method of detecting a proliferative inhibition of lymphocytes in vitro in human amniotic membrane epithelial cells, which is as specifically as follows:

[0062](1) Cascardine epithelial cell vaccination: Take 400 g of cell suspension, centrifuge for 5 min, to the supernatant, add 10% by weight of the gibu blood-free GIBCO 1640 medium heavy, sampling count; adjust the cell concentration to 2 × 105 / ml; the adjusted cell suspension was inoculated into 3 24-well culture plates, and 4 holes were inoculated each, and 12 holes were inoculated with 500 μl per well. Place the inoculated cell plate in a carbon dioxide incubator (CO2The concentration is set to: 5.0 vol%, the temperature is set to: 37.0 ° C), cultured for 20 h;

[0063](2) Subside each well medium in step (1), 500 μL of GIBCO 1640 medium containing 8 μg / ml of silk serum Ceramine Ceramine Ceramine, in the carbon dioxide incubator (CO)2The concentration is set to: 5.0 vol%, the te...

Embodiment 2

[0067]Example 2 of the present invention provides a method of detecting a proliferative inhibition of lymphocytes in vitro in human amniocenteular epithelial cells, which is specifically:

[0068](1) Cascardine epithelial cell vaccination: Take 400 g of cell suspension, centrifuge for 5 min, to the supernatant, add 10% by weight of the gibu blood-free GIBCO 1640 medium heavy, sampling count; adjust the cell concentration to 2 × 105 / ml; the adjusted cell suspension was inoculated into 3 24-well culture plates, and 4 holes were inoculated each, and 12 holes were inoculated with 500 μl per well. Place the inoculated cell plate in a carbon dioxide incubator (CO2The concentration is set to: 5.0 vol%, the temperature is set to: 37.0 ° C), cultured for 20 h;

[0069](2) Subside each well medium in step (1), 500 μl of GIBCO 1640 medium containing 10 wt% fetal bovine serum containing 12 μg / ml of keteng serum is placed in a carbon dioxide incubator (CO) each of the GIBCO 1640 medium containing 1...

Embodiment 3

[0073]Example 3 of the present invention provides a method of detecting a proliferative method of lamb epithelial cells inhibiting lymphocytes, which is specifically:

[0074](1) Cascardine epithelial cell vaccination: Take 400 g of cell suspension, centrifuge for 5 min, to the supernatant, add 10% by weight of the gibu blood-free GIBCO 1640 medium heavy, sampling count; adjust the cell concentration to 2 × 105 / ml; the adjusted cell suspension was inoculated into 3 24-well culture plates, and 4 holes were inoculated each, and 12 holes were inoculated with 500 μl per well. Place the inoculated cell plate in a carbon dioxide incubator (CO2The concentration is set to: 5.0 vol%, the temperature is set to: 37.0 ° C), cultured for 20 h;

[0075](2) Subside each well medium in step (1), 500 μl of GIBCO 1640 medium containing 10 wt% fetal bovine serum containing 10 wt% fetal bovine serum is placed in a carbon dioxide incubator for each GIBCO 1640 medium containing 10 wt% fetal bovine serum Co.2T...

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Abstract

The invention relates to the field of lymphocyte proliferation inhibition, in particular to a method for detecting in-vitro inhibition of lymphocyte proliferation of human amniotic epithelial cells, which comprises the following steps of: co-culturing the human amniotic epithelial cells and stained PBMC (peripheral blood mononuclear cells) in a first complete culture medium in the presence of a free amino stimulant; and recording lymphocyte proliferative capacity. The method for detecting in-vitro inhibition of lymphocyte proliferation of human amniotic epithelial cells is simple to operate and stable in data, the relative standard deviation is within 5%, the inhibition rate is as high as 75%, and the method can be applied to immunoregulation.

Description

Technical field[0001]The present invention relates to the field of lymphocyte proliferation inhibition, and more particularly, the present invention relates to a method of detecting human amniotic epithelial cells inhibiting lymphocyte proliferation inhibitory inhibition.Background technique[0002]The amniocentesis is popular in the form of a transparent film that is wrapped in amniotic fluid, a mature amniotic membrane area of ​​2 square meters, gathered with 300 million amnobium epithelial cells. As the protective layer of the embryo, the amnio is not only providing nutrients for fetal development, but also a wonderful barrier that blocks the mother and the fetus.[0003]The human amniocenteular epithelial cells are present in the amniotic tissue on the placenta after childbirth, and the cell groups are differentiated from the sacral epicocyte groups of fertilized eggs to the 8th day, with differentiation potential, and can differentiate into three epodent. Under suitable induction c...

Claims

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Application Information

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IPC IPC(8): C12Q1/02C12N5/078C12N5/073
CPCG01N33/5047G01N33/5044C12N5/0634C12N2501/515C12N2501/51C12N2502/025
Inventor 朱灏
Owner 华夏源细胞工程集团股份有限公司
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