A preparation method of microfluidic chip embedded with oriented nanofiber membrane

A technology of nanofiber membrane and microfluidic chip, which is applied in the field of microfluidic chip and cell sorting, and can solve the problems of cell damage and low release efficiency

Active Publication Date: 2020-11-13
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, some CTC reversible release systems have made gratifying progress, but there are problems such as cell damage and low release efficiency during the release process.

Method used

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  • A preparation method of microfluidic chip embedded with oriented nanofiber membrane
  • A preparation method of microfluidic chip embedded with oriented nanofiber membrane
  • A preparation method of microfluidic chip embedded with oriented nanofiber membrane

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] (1) Dissolve 37.2mg SH-PEG-FA in 5mL deionized water, add 2.42mg Cysteine ​​Cysteine ​​and 10μL 30% hydrogen peroxide, ice bath magnetic stirring reaction for 1h, and then use the molecular weight cutoff of 1000 The dialysis bag was dialyzed in deionized water (9 times, 2L / time) for three days, and freeze-dried at a vacuum of 0.09 mBar at a low temperature of -45°C for 48 hours to obtain a Cys-PEG-FA aqueous solution.

[0059] (2) Add 0.5g of polyethyleneimine PEI and 1.5g of polyvinyl alcohol PVA into 18g of deionized water, and magnetically stir at 90°C for 4h to obtain a PEI / PVA spinning solution with a mass fraction of 10wt%, which is sucked with a 10mL syringe PEI / PVA spinning solution, the syringe is connected to the 16-gauge needle, the injection pump flow rate is set to 0.6mL / h, the aluminum foil paper wrapped on the rotating shaft of 2500r / min is used as the receiving device, the receiving distance is adjusted to 20cm, and the spinning is set The voltage is 25kV, ...

Embodiment 2

[0066] The present invention is characterized by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR), thermogravimetry (TG) and hemolysis-coagulation test, anti-protein and blood cell adhesion test, and cancer cell capture and release test. The various properties of the functionalized oriented nanofiber membrane and its application potential combined with microfluidic chip in the sorting of circulating tumor cells.

[0067] Fourier transform infrared spectrum test:

[0068] FTIR was used to characterize whether the Cys-PEG-FA containing disulfide bonds was synthesized in step (1) of Example 1, and the results of infrared spectroscopy characterization analysis are as follows: figure 1 As shown, at 2591cm -1 Where is the infrared characteristic absorption peak of sulfhydryl SH in SH-PEG-FA, while the infrared spectrum of Cys-PEG-FA is 2591cm -1 The infrared characteristic absorption peak of sulfhydryl SH disappeared at 456cm -1 The infrared absorption p...

Embodiment 3

[0075] Hemolysis test test:

[0076] The hemolysis test was conducted to study the blood compatibility of the nanofiber membrane before and after modification. Take 1 mL of fresh healthy adult whole blood, centrifuge at 2000 r / min for 5 min, and wash with phosphate buffered saline PBS 3 times to obtain red blood cell HRBCs. Dilute HRBCs 30 times with PBS solution, take out 0.2mL HRBCs dilution and transfer to 1.5mL centrifuge tube containing 0.8mL PBS as a negative control. Take 0.2mL HRBCs diluent and transfer it to a 1.5mL centrifuge tube containing 0.8mL distilled water as a positive control. Subsequently, the configured HRBCs diluent was diluted 5 times with PBS, and the PEI / PVA nanofiber membrane after the crosslinking treatment in step (2) of Example 1 and the PEI / PVA-PMPC-Br nanofiber membrane in step (4) were taken. 4 mg each of the fiber membrane and the PEI / PVA-PMPC-FA nanofiber membrane of step (5) was immersed in 1 mL of the 5-fold diluted HRBCs suspension, and 5 pa...

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Abstract

The invention relates to a production method of an oriented nanofiber membrane-embedded micro-fluidic chip. The method comprises the following steps: carrying out electrostatic spinning and steam crosslinking on a PEI / PVA spinning solution to obtain an oriented PEI / PVA nanofiber membrane; modifying the surface of the oriented PEI / PVA nanofiber membrane with zwitterionic MPC and a targeting ligand Cys-PEG-FA to obtain PEI / PVA-PMPC-FA; and carrying out plasma bonding on the PEI / PVA-PMPC-FA and a herringbone PDMS micro-fluidic chip to obtain the oriented nanofiber membrane-embedded micro-fluidic chip which can be used for sorting circulating tumor cells. The oriented nanofiber membrane-embedded micro-fluidic chip is simple to operate, can rapidly realize the highly-pure capture and rapid release of the circulating tumor cells in blood, endows the nanofiber membrane with excellent anti-protein and anti-blood cell adhesion performances through amphion modification to improve the cell capture purity, and reduces the non-specific adhesion of blood cells through the oriented nanofiber membrane in order to further improve the capture purity, so the oriented nanofiber membrane-embedded micro-fluidic chip has a wide application prospect.

Description

Technical field [0001] The invention belongs to the technical field of microfluidic chips and cell sorting, and particularly relates to a method for preparing a microfluidic chip embedded with an oriented nanofiber membrane. Background technique [0002] The detection of circulating tumor cells (CTC) is of great significance for the early diagnosis of tumors, disease development prediction, curative effect evaluation, prognostic judgment and individualized treatment. However, the number of CTCs in the blood is very small, and it is difficult to achieve capture and separation by conventional means. For example, depending on the density, the separation of various components in blood by density gradient centrifugation is the most classic method. However, CTC may migrate to the plasma layer or remain in red blood cells or neutrophils, causing loss of CTC during the separation process [Lalmahomed ZS, et al. Circulating tumor cells and sample size: the more, the better. Journal of Cli...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12N5/09D06M15/356D06M13/10D06M15/53D04H1/4382D06B13/00D06M101/24D06M101/30
CPCC12M23/16C12N5/0693D04H1/4382D06B13/00D06M13/10D06M15/356D06M15/53D06M2101/24D06M2101/30
Inventor 史向阳肖云超王梦媛
Owner DONGHUA UNIV
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