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Serialized perfusion type microfluidic device

A microfluidic device and perfusion technology, applied in the field of biomedicine, can solve problems such as difficult to achieve cryoprotectant treatment, and achieve the effect of avoiding the accumulation effect of osmotic pressure difference and reducing osmotic damage

Active Publication Date: 2020-02-11
UNIV OF ELECTRONICS SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, in the membrane microfluidic system introduced by Lusianti and Higgins, both simulation and experiment prove that it is difficult to achieve sufficient cryoprotectant treatment (Biomicrofluidics, 2014, 8, 054124)

Method used

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  • Serialized perfusion type microfluidic device
  • Serialized perfusion type microfluidic device
  • Serialized perfusion type microfluidic device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] see figure 1 with figure 2 , a serialized perfusion microfluidic device comprising a plate-shaped perfusion-side body 1 , a plate-shaped middle layer body 3 and a plate-shaped cell-side body 2 arranged in sequence from top to bottom.

[0041] The materials of the body 1 on the perfusion side, the body 2 on the cell side and the body 3 on the middle layer are the same, all being polymethyl methacrylate (PMMA). The thickness of the perfusion side body 1 is 10 mm, the thickness of the cell side body 2 is 5 mm; the thickness of the middle layer body 3 is 0.1 mm.

[0042] The perfusion side body 1 is provided with a perfusion side inlet 11, and the inner surface of the perfusion side body 1 corresponding to the three membrane windows on the middle layer body 3 is provided with a perfusion side flow channel 12. The total length of the flow channel 12 is 300mm, width 1mm, and depth 0.5mm, one end of the perfusion side channel 12 is connected to the perfusion side inlet 11;

...

Embodiment 2

[0047] Add cryoprotectant to stem cell suspension.

[0048] The commonly used cryoprotectant for stem cell cryopreservation is DMSO with a concentration of 10%. To add DMSO to the cell suspension to this target concentration, the following setup was used: a 20% DMSO solution was used as the perfusate, and the initial concentration of DMSO in the stem cell suspension was 0%. The specific form of setting the serialized perfusion microfluidic device is as described in Example 1. During the treatment process, the flow rate of the cell suspension at the cell side inlet 21 is 1ml / min, which is injected from the cell side inlet 21; the perfusate at the perfusion side inlet 11 The flow rate is the same as 1ml / min, which is injected from the inlet 11 on the perfusion side; the perfusate of the injection device flows into the stem cell suspension through the first membrane window 31, the second membrane window 32, and the third membrane window 32, and finally flows through the cell side...

Embodiment 3

[0056] Removal of cryoprotectants from stem cell suspensions

[0057] The device of the present invention can be used to remove the cryoprotectant in the stem cells with physiological saline as the perfusate. Pass the cell suspension containing the cryoprotectant through the inlet 21 of the cell fluid side, and set the inlet flow rate to 0.5ml / min; feed the perfusate into the perfusate inlet 11, and set the inlet flow rate to 5ml / min; In 23 places, the cell suspension with a fixed dilution ratio can be obtained, and the removal rate of the cryoprotectant in the cells is above 90%.

[0058] Below in conjunction with accompanying drawing, the principle of present embodiment 3 is described in further detail:

[0059] join Figure 7 In embodiment 3, the reverse flow of flow channels on both sides of the device forms transmembrane pressures that increase sequentially on both sides of the three membrane windows, and the principle is the same as in embodiment 2.

[0060] see Fig...

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Abstract

The invention relates to a serialized perfusion type microfluidic device. The serialized perfusion type microfluidic device comprises a plate-shaped perfusion side body, a plate-shaped intermediate layer body and a plate-shaped cell side body which are arranged from top to bottom in sequence; the perfusion side body is provided with a perfusion side flow passage and a perfusion side inlet, the intermediate layer body is provided with three membrane windows or above, and the membrane windows are filled with microfiltration membranes; the cell side body is separately provided with a cell side inlet, a cell side flow passage and a cell side outlet which are sequentially connected; the perfusion side inlet and the cell side outlet are correspondingly positioned at the same side of the device from top to bottom, and the cell side inlet is positioned at the other side of the device; when the device works, perfusate is injected into the perfusion side inlet, and cell suspension is injected into the cell side inlet; the perfusate flows into the cell side flow passage in an one-way mode through the membrane windows, the perfusate is mixed with the cell suspension, and the mixture is discharged from the cell side outlet. The device can avoid an accumulation effect of intracellular and extracellular osmotic pressure difference in continuous type treatment systems, including a diffusion method, a dialysis method, etc., thereby better adapting to high-throughput treatment requirements.

Description

technical field [0001] The invention relates to the field of biomedicine and is used for cell suspension treatment, especially for adding or removing cryoprotectant to the cell suspension. Background technique [0002] Biomedicine often involves changing the concentration of a certain component of a cell suspension. Due to the selective passage characteristics of the cell membrane, changes in the concentration of external solutes will cause a difference in osmotic pressure between the inside and outside of the cell, resulting in a change in the osmotic volume of the cell. Especially when this change is too fast, it often causes osmotic damage to cells. This phenomenon is especially evident in the process of adding or removing cryoprotectant during cryopreservation. [0003] Low temperature preservation is widely used in the long-term preservation of rare blood red blood cells, stem cells, immune cells and other living materials. In order to avoid or reduce low temperature...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/06C12M3/02
CPCC12M23/16C12M29/04C12M29/10
Inventor 周晓明蒋志豪刘杰王吉李亚东
Owner UNIV OF ELECTRONICS SCI & TECH OF CHINA
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