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A 3D printing-based T lymphocyte expansion method

A lymphocyte and 3D printing technology, which is applied in the field of T lymphocyte expansion, can solve the problem of limited space for cell proliferation, and achieve an effective expansion path and a short time-consuming effect

Active Publication Date: 2022-07-12
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention provides a T lymphocyte expansion method, which can solve the technical problems of the traditional T lymphocyte expansion method that the cell proliferation space is limited and cannot form continuous and controllable stimulation for cell proliferation, thereby realizing rapid expansion of T lymphocytes. Lymphocytes

Method used

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  • A 3D printing-based T lymphocyte expansion method
  • A 3D printing-based T lymphocyte expansion method
  • A 3D printing-based T lymphocyte expansion method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] 1. Preparation of functionalized bioink

[0047] like figure 1 As shown in the figure, GelMA (w / v, 10%) microspheres were prepared by emulsification method, and GelMA microspheres with a diameter of about 1 μm were selected, CD3 antibody and CD28 antibody were coupled on the surface, and IL-2 molecules were loaded inside, and the IL-2 concentration was 300U / mL, CD3 antibody and CD28 antibody concentration were 10ng / mL, respectively.

[0048] The number of microcarriers in the bioink is adjusted according to the number of T lymphocytes, and the ratio of microcarriers to T lymphocytes is maintained at 1:1.

[0049] 2. Mix T lymphocytes with the functionalized bioink, and control the concentration of T lymphocytes in the mixed system to be 1×10 6 / mL;

[0050] 3D printing was carried out, solidified and cross-linked while printing, and then cultured in the G-Rex gas permeable culture system for 12 days; the medium used was a special medium for lymphocyte culture.

Embodiment 2

[0052] Compared with Example 1, the only difference is that the initial concentration of T lymphocytes in the bioink is 2.5 × 10 6 / mL.

Embodiment 3

[0054] Compared with Example 1, the only difference is that the initial concentration of T lymphocytes in the bioink is 0.5×10 6 / mL.

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Abstract

The invention relates to a T lymphocyte expansion method, in particular to a T lymphocyte expansion method based on 3D printing. The method includes providing a functionalized bioink; the functionalized bioink includes a bioink body, and a microcarrier combined with IL-2, CD3 antibody and CD28 antibody; T lymphocytes are combined with the functionalized bioink The 3D printed T lymphocyte-containing 3D structures were cultured. The method of the invention can solve the technical problems that the traditional T lymphocyte expansion method has limited space for cell proliferation and cannot form a continuous and controllable stimulation for cell proliferation, thereby realizing rapid expansion of T lymphocytes.

Description

technical field [0001] The invention relates to a T lymphocyte expansion method, in particular to a T lymphocyte expansion method based on 3D printing. Background technique [0002] The activation and proliferation of T lymphocytes in vivo requires the joint participation of the first signal, the second signal and the third signal, wherein the first signal refers to the T lymphocyte surface receptor (TCR) and the target cell surface histocompatibility antigen (MHC) binding elicited stimulatory signals, secondary signals refer to co-stimulatory molecules (e.g., CD28-B7 molecule pair, B7 expressed on the surface of antigen-presenting cells (APC), CD28 expressed on the surface of T lymphocytes) Stimulatory signals, the third signal refers to cytokines involved in the regulation of this process. [0003] As adoptive T lymphocyte immunotherapy (eg, genetically engineered CAR-T and TCR-T cell therapy) continues to create great success stories in preclinical and clinical studies, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2501/2302C12N2501/515C12N2501/51C12N2513/00C12N2533/54C12N2533/80C12N2533/72C12N2533/30C12N2533/50
Inventor 孙伟庞媛周珍珍
Owner TSINGHUA UNIV