Cultivation method for differentiating into T cell of embryonic stem cell and differentiation culture medium

A technology of embryonic stem cells and culture methods, applied in the field of culture of embryonic stem cells differentiated into T cells, can solve the problems of carcinogenesis, teratogenicity, difficulty in clinical application, cell mutation, etc.

Active Publication Date: 2007-10-31
SUN YAT SEN UNIV
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Problems solved by technology

However, the reported induction systems all have the following two defects: (1) they must rely on exogenous cells to provide the three-dimensional support and differentiation signals required for T lymphocyte differentiation. Some precursor cells of the cells are very similar, and some cells are easily mutated by the transfer of foreign genes, thus limiting the application of T cells
(2) The carrier used when transferring the foreign gene has the risk of carcinogenesis and teratogenicity, so it is difficult to enter the actual clinical application

Method used

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  • Cultivation method for differentiating into T cell of embryonic stem cell and differentiation culture medium

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Embodiment Construction

[0022] Materials: Embryonic stem cells (embryonic stem cells, ES cells) are the international common cell line E14.1 and the cell lines SC1001 and SC1002 derived from B6 / 129 established by the inventor's laboratory.

[0023] Instruments mainly include: CO2 incubator, centrifuge, flow cytometer, etc.

[0024] The method for culturing embryonic stem cells differentiated into T cells according to the present invention comprises the following steps:

[0025] A. Preparation of embryoid bodies: Embryonic stem cells were digested into single cells with trypsin and inoculated in high-glucose DMEM medium containing 15% fetal bovine serum for suspension culture for 5-7 days to induce the formation of embryoid bodies.

[0026] The high-sugar DMEM medium is selected from the CAT NO.SH30003.01 product of HYCLONE Company, or similar products of GIBCO Company.

[0027] B. Embryonic bodies are transferred to differentiation culture medium, and described differentiation culture medium is to a...

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Abstract

The culture method to differentiate embryonic stem cell into T cell comprises: preparing embryo body; transferring the body into differentiated culture medium; digesting the body into single cell by pancreatin; labeling the opposite antibody of T cell special surface cell; detecting T cell quantity with a FCM; and obtaining the need T cell. It also provides the opposite culture medium. This invention overcomes the defect in prior art, induces T lymphocyte from embryonic stem cell directly, and provides a simple and effective external model for research and a safe source for clinical application of T cell.

Description

technical field [0001] The invention belongs to the field of stem cells and immunology, and relates to a method for culturing stem cells, in particular to a method for culturing embryonic stem cells differentiated into T cells. Background technique [0002] T lymphocytes are one of the main effector cells of specific immune response, and play an important role in infectious diseases, especially viruses and intracellular parasites, as well as tumor immunity. The abnormality of T lymphocytes can not only make the host susceptible to infectious diseases and tumors, but also lead to autoimmune diseases and allergic diseases. Common clinical transplant rejection and many senile diseases are also closely related to T lymphocytes. [0003] So far, the development process and mechanism of T lymphocytes are not very clear. This is mainly because the development of T lymphocytes must rely on the thymus microenvironment, therefore, it is difficult to induce T lymphocytes in vitro, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/06C12N5/08C12N5/0783
Inventor 项鹏李树浓彭延文
Owner SUN YAT SEN UNIV
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