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T lymphocyte amplification method based on 3D printing

A lymphocyte, 3D printing technology, applied in the field of T lymphocyte expansion, can solve the problem of limited space for cell proliferation, and achieve the effect of promoting rapid expansion, short time-consuming, and effective T lymphocyte expansion pathway

Active Publication Date: 2020-08-14
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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  • T lymphocyte amplification method based on 3D printing
  • T lymphocyte amplification method based on 3D printing
  • T lymphocyte amplification method based on 3D printing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] 1. Preparation of functionalized bioink

[0047] like figure 1 As shown, GelMA (w / v, 10%) microspheres were prepared by emulsification method, GelMA microspheres with a diameter of about 1 micron 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 concentrations were 10ng / mL.

[0048] The number of microcarriers in the bioink was adjusted according to the number of T lymphocytes, and the ratio of microcarriers to T lymphocytes was kept 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] Carry out 3D printing, solidify the cross-linking while printing, and then culture in the G-Rex gas-permeable culture system for 12 days; the medium used is 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 amplification method, in particular to a T lymphocyte amplification method based on 3D printing. The method includes the following steps: providing a functionalized bio-ink, wherein the functionalized bio-ink comprises a bio-ink body and a micro-carrier combined with IL-2 and CD3 antibodies and a CD28 antibody; mixing T lymphocytes with the functionalized bio-ink, and carrying out 3D printing; and culturing the 3D-printed 3D structure containing the T lymphocytes. The method disclosed by the invention can solve the technical problems that the cell proliferation space is limited and continuous and controllable stimulation cannot be formed on cell proliferation in traditional T lymphocyte proliferation methods, and realizes rapid proliferation of T lymphocytes.

Description

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

Claims

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Application Information

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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
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