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Method for promoting in-vitro amplification of umbilical cord blood T cell and maintaining high-proportion TSCM subpopulation

A technology of umbilical cord blood and cell bodies, applied in the field of cytology

Inactive Publication Date: 2019-06-28
四川药智联恒科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, whether low-concentration CAG can also increase the proliferation of T cells derived from umbilical cord blood has not been reported yet.

Method used

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  • Method for promoting in-vitro amplification of umbilical cord blood T cell and maintaining high-proportion TSCM subpopulation
  • Method for promoting in-vitro amplification of umbilical cord blood T cell and maintaining high-proportion TSCM subpopulation
  • Method for promoting in-vitro amplification of umbilical cord blood T cell and maintaining high-proportion TSCM subpopulation

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Embodiment 1

[0023] 1 Materials and methods

[0024] 1.1 Main reagents and instruments

[0025] Cycloastragenol (HPLC≥98%) was purchased from Chengdu Mansite Biological Company. RPMI1640 medium was purchased from Hyclone Company. Fetal Bovine Serum (FBS) was purchased from BOVOGEN Company. Lymphoprep lymphatic separation liquid was purchased from Axis-shield Company. Penicillin-streptomycin double antibody was purchased from Gibco. Interleukin 7 (IL-7) and interleukin 15 (IL-15) were purchased from Cellgenix. Dynabeads Human T-Activator CD3 / CD28 was purchased from Gibco. FITC anti-humanCD45RA, APC / cy7 anti-humanCD8, PE / cy7 anti-humanCD4, PE anti-humanCD197 (CCR7), PE anti-humanCD3, and CFSE were purchased from BD Company. ACEA NovoCyte series flow cytometer (ACEA NovoCyte 3130) was purchased from ACEA Biosciences. Low speed centrifuge, CO 2 The incubator was purchased from Thermo Fisher Company.

[0026] 1.1 1.2 Separation of Umbilical Cord Blood Mononuclear Cell (UCBMC)

[0027] H...

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Abstract

The invention discloses a method for promoting in-vitro amplification of an umbilical cord blood T cell and maintaining a high-proportion TSCM subpopulation. The method comprises the steps that cycloastragenol, a cell factor IL-7 and a cell factor IL-15 are combined and added into an in-vitro culture solution of the T cell to promote the amplification of the T cell and maintain the high-proportionTSCM subpopulation. The method for promoting the in-vitro amplification of the umbilical cord blood T cell and maintaining the high-proportion TSCM subpopulation has the advantages that the high-proportion TSCM cell subpopulation can be maintained while the low-concentration CAG combined with the IL-7 and the IL-15 to promote the amplification of the umbilical cord blood derived T cell is successively explored; the novel in-vitro culture method lays an experimental foundation for the in-vitro long-term culture of the T cell and an immunization therapy containing a CAR-T cell therapy.

Description

technical field [0001] The invention belongs to the field of cytology, and in particular relates to a method for promoting the in vitro expansion of umbilical cord blood T cells and maintaining a high proportion of T cells SCM subgroup method. Background technique [0002] Immune cell therapy is the most promising tumor treatment method, including tumor-infiltrating lymphocyte (TIL) therapy, T cell receptor engineered-T cell (TCR-T) therapy and Chimeric Antigen Receptor T-cell (Chimeric Antigen Receptor Engineered T-cell, CAR-T) therapy, etc. Among them, TIL therapy is a treatment method in which tumor-infiltrating lymphocytes are isolated from the patient's tumor tissue, and after in vitro expansion, the cell population with tumor cell killing activity is returned to the patient's body. It is currently the most effective cell therapy for melanoma; Both TCR-T therapy and CAR-T therapy use genetic modification technology to improve the ability of T cells to specifically rec...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
Inventor 王晓东郑武燕
Owner 四川药智联恒科技有限公司
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