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Detection method of immune functions of mesenchymal stem cells

A technology of high-quality stem cells and immune function, which is applied in the field of stem cell application to achieve the effect of good quality and strong immune function

Pending Publication Date: 2020-05-15
深圳市芙丽嘉科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for mesenchymal stem cells, a method that can efficiently detect its immune function has not been found

Method used

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  • Detection method of immune functions of mesenchymal stem cells
  • Detection method of immune functions of mesenchymal stem cells
  • Detection method of immune functions of mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Detection of lymphocyte proliferation activity

[0027] 1. Isolation and culture of human umbilical cord MSCs

[0028] The umbilical cord tissue of the postpartum full-term fetus was collected, and the umbilical cord tissue was processed, and the umbilical cord MSCs were obtained by the adherent culture method or enzyme digestion culture method.

[0029] 2. Human Umbilical Cord MSCs Processing

[0030] ① When the confluence of MSCs reaches 90% in the T75 flask, discard the old medium, add stem cell medium containing 10 μg / mL mitomycin C, and put it back into the incubator for 2 hours. Then the mitomycin C medium was discarded, and the cells were washed 4 times with normal saline to fully wash away the mitomycin C.

[0031] ②Then add 0.125% trypsin to digest the stem cells for 1 min, and finally add complete medium containing 10% fetal bovine serum (FBS) to terminate the digestion, and pipette repeatedly to make it into a single cell suspension.

[0032] ③ Collect the...

Embodiment 2

[0042] Lymphocyte subset detection

[0043] Steps 1-3 are as in Example 1.

[0044] 4. Spread mitomycin C-treated stem cells on a 24-well plate according to the cell density, and the number of fixed PBMCs in each well is 2×10 6 cells / well, MSCs: PBMCs cell number ratio (0:1), (1:10), (1:50) MSC cells were plated, and the well plate was placed in CO 2 incubator.

[0045] 5.1 Detection of Treg cell population

[0046] Detection of CD4 by Flow Cytometry + CD25 + FOXP3 + The cell population is Tregs cells.

[0047] The result is as figure 2 shown. figure 2 It is the detection result of the lymphocyte Tregs cell population of the method for detecting the immune function of mesenchymal stem cells according to an embodiment of the present invention. from figure 2 As can be seen in the human umbilical cord MSCs on CD4 + CD25 + FOXP3 + The inhibitory effect of the cell population was enhanced with the increase of the concentration, and the results of P2 and P5 MSCs showed...

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Abstract

The invention provides a detection method of immune functions of mesenchymal stem cells. The detection method of the immune functions of the mesenchymal stem cells comprises the following steps: (1) co-culturing umbilical cord mesenchymal stem cells and peripheral blood mononuclear cells so as to obtain a sample to be tested; and (2) detecting proliferative activity of lymphocytes and proportion of lymphocyte subsets in the sample to be tested. The detection method of the immune functions of the mesenchymal stem cells is suitable for non-diagnostic purposes. According to the detection method of the immune functions of the mesenchymal stem cells provided by the embodiment of the invention, the mesenchyma stem cells and the human peripheral blood mononuclear cells are co-cultured so as to obtain the sample to be tested; and then, the proportion of the lymphocyte subsets and the proliferative activity of the lymphocytes in the sample to be tested are detected so as to evaluate the immunoregulatory capacity of the mesenchymal stem cells. The greater the changes of the proportion of the lymphocyte subsets and the proliferative activity of the lymphocytes are, the stronger the immune functions and the better the quality of the mesenchymal stem cells are.

Description

technical field [0001] The invention relates to the technical field of stem cell application, in particular to a method for detecting the immune function of mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells are a group of cells first isolated from bone marrow in the 1970s. Later, people found through research that this kind of cells has multi-directional differentiation potential, and can differentiate into osteoblasts, chondrocytes, adipocytes, cardiomyocytes, etc. under certain induction culture conditions. The differentiation potential can still be maintained. Because they can differentiate into a variety of mesenchymal stem cells derived from mesoderm, they are called mesenchymal stem cells (MSCs). In the 1990s, people isolated mesenchymal stem cells from human umbilical cord tissue. After research, it was found that umbilical cord mesenchymal stem cells met the definition of mesenchymal stem cells by the International Society for Cellular Th...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
CPCG01N33/5008G01N33/505
Inventor 陶伟贺光锐王慧琴
Owner 深圳市芙丽嘉科技有限公司
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