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Construction method and application of stem cell quantitative standard for screening immunomodulatory ability of high-quality human umbilical cord mesenchymal stem cells

A quality stem cell and immune regulation technology, which is applied in the field of screening high-quality human umbilical cord mesenchymal stem cells and the construction of a quantitative standard for stem cells with immune regulation ability, can solve problems such as the lack of effective detection methods for MSCs immune regulation, and achieve the effect of improving the therapeutic effect.

Pending Publication Date: 2021-06-22
NANJING DRUM TOWER HOSPITAL
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Problems solved by technology

[0004] The invention provides a construction method and application of a stem cell quantification standard for screening high-quality human umbilical cord mesenchymal stem cells for immunoregulation ability, so as to overcome the technical problem of lack of effective methods for detecting MSCs immunoregulation at present

Method used

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  • Construction method and application of stem cell quantitative standard for screening immunomodulatory ability of high-quality human umbilical cord mesenchymal stem cells
  • Construction method and application of stem cell quantitative standard for screening immunomodulatory ability of high-quality human umbilical cord mesenchymal stem cells
  • Construction method and application of stem cell quantitative standard for screening immunomodulatory ability of high-quality human umbilical cord mesenchymal stem cells

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Embodiment 1

[0049] This embodiment provides a method for constructing quantitative standards for screening high-quality human umbilical cord mesenchymal stem cells for immunoregulatory ability stem cells, including the following steps:

[0050] Step 1: Primary isolation and expansion of umbilical cord mesenchymal stem cells from 12 strains of human umbilical cord, and subculture to P5 generation. The 12 strains of umbilical cord mesenchymal stem cells are umbilical cord mesenchymal stem cells of different genders and ages.

[0051] Among them, the isolation and expansion methods of human umbilical cord mesenchymal stem cells are as follows: collect human umbilical cord under sterile conditions, clean umbilical cord blood, remove umbilical arteries and veins, cut into pieces, adherent culture, digestion, centrifugation, resuspension, inoculation Culture until 80-90% confluence, cell digestion, subculture, expansion culture, and the umbilical cord mesenchymal stem cells in the logarithmic p...

Embodiment 2

[0093] This example provides an application of the quantification standard for screening high-quality human umbilical cord mesenchymal stem cells for immunoregulatory ability: according to the requirement of immunoregulatory ability of stem cells transplanted into the body, detect and calculate the Th1 inhibitory proliferation ratio and Th17 inhibition of the stem cells to be screened Proliferation ratio or Treg proliferation promotion ratio; compare it with the corresponding quantification standard, and the stem cells greater than the quantification standard are high-quality human umbilical cord mesenchymal stem cells that can be screened for immunoregulatory ability.

[0094] Wherein, the method for detecting and calculating the Th1 inhibitory proliferation ratio and the Th17 inhibitory proliferation ratio of the stem cells to be screened is: co-culture the stem cells to be screened with freshly isolated human peripheral blood mononuclear cells (PBMCs) as the Th-MSC group; In...

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Abstract

The invention discloses a construction method and application of a stem cell quantitative standard for screening immunomodulatory ability of high-quality human umbilical cord mesenchymal stem cells. The construction method comprises the following steps: step 1, carrying out primary isolation and amplification on umbilical cord mesenchymal stem cells from a plurality of human umbilical cords, and subculturing to P5 generation; step 2, carrying out flow analysis on the Th1 subgroup and the Th17 subgroup of the lymphocyte subgroup and carrying out flow analysis on the regulatory T cell subgroup of the lymphocyte subgroup; 3, step analyzing a streaming result obtained in the step 2 by adopting Flowjo software, and calculating the proliferation inhibition ratio of Th1, the proliferation inhibition ratio of Th17 and the proliferation promotion ratio of Treg of each group; and respectively averaging to obtain a corresponding stem cell quantitative standard for screening the immunomodulatory ability of the high-quality human umbilical cord mesenchymal stem cells. The invention provides a cell selection standard for basic research and clinical transformation of stem cells for treating autoimmune diseases and immune disorder related diseases, and improves the treatment effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a quantitative standard for screening stem cells, in particular to a construction method and application of a quantitative standard for screening high-quality human umbilical cord mesenchymal stem cells with immune regulation ability stem cells. Background technique [0002] Mesenchymal Stem Cells (MSCs) can be isolated and cultured from a series of tissues, including but not limited to bone marrow, fat, umbilical cord, amniotic membrane, and placenta. Multi-source cells of various mesenchymal tissue cells such as adipocytes, osteoblasts, chondrocytes, hepatocytes and nerve cells. Therefore, mesenchymal stem cells have become a research hotspot in the field of basic medicine and clinical tissue and organ injury repair and regeneration. [0003] Among them, human umbilical cord derived MSCs (human umbilical cord derived MSCs, HUCMSCs) have the advantages of a wide range of sources, non...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775G01N15/14G01N33/68
CPCC12N5/0665G01N15/14G01N33/6866G01N33/6869C12N2509/00C12N2509/10G01N2333/57G01N2333/54
Inventor 王斌谢园园
Owner NANJING DRUM TOWER HOSPITAL
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