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Upper-layer chip and micro device for multistage addition of cell cryoprotectant

A cryoprotectant and chip technology, applied in the field of medical devices, can solve the problems of dilution, easy loss of cells, low addition efficiency, etc., and achieve the effects of reducing mechanical damage, reducing osmotic pressure damage, and improving adding efficiency.

Pending Publication Date: 2021-02-23
GUANGDONG PROV MEDICAL INSTR INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are microfluidic devices based on the principle of laminar interface diffusion and the principle of micromembrane dialysis to realize the addition of precious micro-cell cryoprotectants, but the addition efficiency is low, and cells are easily lost and diluted

Method used

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  • Upper-layer chip and micro device for multistage addition of cell cryoprotectant
  • Upper-layer chip and micro device for multistage addition of cell cryoprotectant
  • Upper-layer chip and micro device for multistage addition of cell cryoprotectant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055]The first processing unit in the cryoprotectant additive liquid generating area 6 is provided with four first mixers, and the diluent and the cryoprotectant stock solution are combined in the channel of the cryoprotectant additive liquid generating area 6 at a certain flow ratio and are in the first A mixer is uniformly mixed, and then a part of the diluted cryoprotectant solution produced enters the cryoprotectant stepwise addition zone 8 to perform the addition of the cryoprotectant, and the other part of the diluted cryoprotectant solution and the diluent are then lowered at a certain flow ratio. The mixing is carried out in the first mixer of the first stage, and the production of four gradient concentrations of cryoprotectant additives with volume fractions of 12.5%, 10%, 7.5%, 5% DMSO and flow rates of 0.1 mL / min is realized step by step.

[0056]The cryoprotectant stepwise addition zone 8 is equipped with 4 second mixers and cell separation and concentration modules. The c...

Embodiment 2

[0059]Add fetal bovine serum to DMEM medium and mix well to obtain a dilution containing 40% (mass to volume ratio) of fetal bovine serum. Treat cultured cells MCF-7 to obtain 1×106Cells / mL of cell suspension. The diluent, cryoprotectant stock solution (100% DMSO), and cell suspension were injected into the microbe at 0.365 mL / min, 0.035 mL / min, and 0.1 mL / min from the first inlet 1, the second inlet 2, and the third inlet 3, respectively. Device.

[0060]The diluent and the cryoprotectant stock solution flow along the channel of the cryoprotectant additive liquid producing area 6, and when they reach the mixer, they are evenly mixed, and gradually form 12.5%, 10%, 7.5%, 5% DMSO cryoprotectant additive liquid And each flow into the cryoprotectant stepwise addition zone 8 at a flow rate of 0.1 mL / min. Subsequently, the 0.1mL / min cell suspension was successively combined with the cell cryoprotectant additive solution 5%, 7.5%, 10%, 12.5% ​​DMSO in the cryoprotectant stepwise addition zon...

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Abstract

The invention discloses an upper-layer chip and a micro device for multistage addition of a cell cryoprotectant. The upper-layer chip is provided with a cryoprotectant addition liquid generation areaand a cryoprotectant step-by-step addition area, and the cryoprotectant addition liquid generation area comprises a first liquid inlet channel and a first processing unit; the first processing unit comprises a plurality of first mixers which are connected in series; the cryoprotectant step-by-step addition area comprises a plurality of second treatment units which are connected in series; and thefirst mixers and the second processing units are arranged in a reverse pairing manner. The first treatment unit is provided with multiple stages of first mixers for diluting to obtain a cryoprotectantadditive solution with gradient concentration, and then the cryoprotectant additive solution flows into the cryoprotectant step-by-step addition area in a graded manner to be mixed with the cell suspension in a graded manner, separated and concentrated, and the cryoprotectant is added step by step so that mechanical damage and osmotic pressure damage of cells are reduced; cell dilution is avoided, addition efficiency is improved and cost is reduced. The upper-layer chip and the micro device can be widely applied to the technical field of medical instruments.

Description

Technical field[0001]The present invention relates to the field of medical devices, and in particular, to a multi-stage additional cell cryopreservation protector.Background technique[0002]Low temperature preservation is an important technique for precious cell preservation in the fields of reproductive medical, tissue regeneration and cellular treatment. Low-temperature protective agents (such as dimethyl sulfoxide DMSO, glycerol, glycerol, etc.) enable cells to avoid low temperature damage during low temperature storage, improve the survival ability after cell thaw, so the cells need to be added to low temperature protection before storage Agent. However, during the low temperature protection agent added, the cells can severe osmotic injury due to the imbalance of intracellular osmosis pressure. Therefore, it is extremely important to safely and effectively load a low temperature protection agent into a cell sample.[0003]Traditionally, a step-by-step loading method is often loaded...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0242
Inventor 邹丽丽龚尧伊翔陈龙胜
Owner GUANGDONG PROV MEDICAL INSTR INST
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