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Integrated micro-fluidic chip device and method for separating T cells and amplifying integrated gel incubation droplets

A microfluidic chip and cell technology, applied in tissue cell/virus culture devices, biochemical equipment and methods, biological material sampling methods, etc., can solve the problem of high differentiation ability of T cells, difficult separation of T cells, T cells Low activity and other problems, to achieve high amplification efficiency, improve effectiveness, and easy operation

Active Publication Date: 2022-01-04
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the existing technology exists: it is difficult to quickly isolate T cells, the activity of isolated T cells is low, CD8 + Hyperdifferentiation and underpotency of T cells

Method used

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  • Integrated micro-fluidic chip device and method for separating T cells and amplifying integrated gel incubation droplets
  • Integrated micro-fluidic chip device and method for separating T cells and amplifying integrated gel incubation droplets
  • Integrated micro-fluidic chip device and method for separating T cells and amplifying integrated gel incubation droplets

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0048] As a preferred embodiment, the confluence connecting channel 33 includes a second serpentine channel 331 and a vertical connecting channel 332;

[0049] One end of the second serpentine channel 331 communicates with the T cell pipeline 26, and the other end vertically communicates with the end of the vertical connecting channel 332 close to the separation pipeline 2;

[0050] One end of the oil phase channel 32 is in communication with the oil phase inlet 31 , and the other end of the vertical connection channel 332 is in vertical communication with the end away from the separation pipeline 2 .

[0051] The function of the second serpentine channel 331 is to stabilize the fluid in the channel, which is beneficial to the subsequent formation of droplets. Then through the vertical connecting channel 332, because the ends of the oil phase channel 32 in the vertical connecting channel 332 are vertically connected to form an inverted T-shaped structure, the continuous fluid ...

Embodiment 1

[0074] Example 1. An integrated microfluidic chip device for separating T cells and amplifying integrated gel incubation droplets and its preparation method

[0075] 1. An integrated microfluidic chip device for separating T cells and expanding integrated gel incubation droplets, such as figure 1 shown, including:

[0076] A chip body 1, including a substrate 11 and a cover sheet 12, the chip body is 35mm long and 25mm wide;

[0077] And the flow channel arranged in the chip body; the flow channel includes:

[0078] The separation pipeline 2 includes a buffer inlet 21, a blood inlet 22, a spiral channel 23, a first separation pipeline 24, a second separation pipeline 25, a T cell pipeline 26, a first waste liquid collection hole 27 and a second waste liquid Liquid collection hole 28, the buffer solution inlet 21 and the blood inlet 22 communicate with the beginning of the spiral channel 23 respectively, the first separation pipeline 24, the second separation pipeline 25 and ...

Embodiment 2

[0086] Example 2. Preparation of T cell gel incubation droplets using a chip

[0087] The chip device described in Example 1 was sterilized by ultraviolet light for use. Before preparing the cell vaccine, 1% BSA solution was passed into the chip to reduce the adhesion of cells and channel components. Blood was collected from the orbital venous plexus of mice, and the blood was stored in a blood collection tube, which was pre-added with sodium heparin as an anticoagulant. Buffer containing 50mM alginate, 50mM Ca-EDTA complex, TLR1 / 2 agonist (1g / mL), cytokine IL-21 (10ng / mL), anti-human CD3 antibody (1g / mL) and Anti-human CD28 antibody (1 g / mL) in PBS buffer.

[0088] Such as figure 1 As shown in (A), the buffer solution inlet 21 and the blood inlet 22 were respectively connected to syringe pumps, and the samples were simultaneously driven at a constant flow rate. The flow rates are Qv1 (blood) = 20 μL / min, Qv2 (buffer) = 200 μL / min. After the separated part is stabilized (ab...

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Abstract

The invention discloses an integrated micro-fluidic chip device and method for separating T cells and amplifying integrated gel incubation droplets. The device comprises a chip body, a separation pipeline arranged in the chip body and a split charging pipeline, the separation pipeline comprises a buffer solution inlet, a blood inlet, a spiral channel, a first separation pipeline, a second separation pipeline, a T cell pipeline, a first waste liquid collection hole and a second waste liquid collection hole, the buffer solution inlet and the blood inlet are correspondingly in communication with the initial end of the spiral channel; the first separation pipeline, the second separation pipeline and the T cell pipeline are correspondingly in communication with the tail end of the spiral channel; the split charging pipeline comprises an oil phase inlet, an oil phase channel, a convergence connecting channel, a first serpentine channel and a gel incubation droplet collecting hole; and one end of the convergence connecting channel is in communication with the T cell pipeline, and the other end of the convergence connecting channel is vertically in communication with the tail end of the oil phase channel to form an inverted T-shaped structure. The separable T cells according to the present invention maintain low differentiation and high potency while proliferating rapidly.

Description

technical field [0001] The present invention relates to the interdisciplinary technical field of microfluidic technology and cancer immunotherapy, in particular to an integrated microfluidic chip device and method for separating T cells and amplifying integrated gel incubation droplets. Background technique [0002] In recent years, immunotherapy has emerged as a powerful and potential treatment for various types of cancer and recurrent viral diseases. Adoptive cell transfer (ACT) is a type of immunotherapy that involves the isolation and expansion of antigen-specific T cells in vitro, followed by adoptive transfer back to the patient. Accumulating evidence indicates that antigen-specific CD8 + Adoptive transfer of T cells may be an effective strategy against chronic viral infections and malignancies such as melanoma. A recent study by Rosenberg SA showed that circulating PD-1 in peripheral blood + CD8 + T cells can specifically recognize melanoma antigens and bind to tu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12N5/0783
CPCC12M23/16C12M33/10C12M25/01C12N5/0636Y02A50/30
Inventor 张志凌田已申
Owner WUHAN UNIV
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