Fetal cell capturing module, application method thereof, micro-fluidic chip for fetal cell capturing and application method of micro-fluidic chip for fetal cell capturing

A microfluidic chip and fetal cell technology, applied in the field of cell capture, can solve the problems of low capture efficiency, high cost and low purity of fetal cell analysis

Active Publication Date: 2020-07-24
WISDOM HEALTHY BIOTECH XIAMEN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of low capture efficiency, low purity, high cost and difficulty in genome-wide analysis of existing fetal cell analysis technology, the following fetal cell capture module is proposed for Microfluidic chips for capturing fetal cells and their respective usage methods to achieve high-efficiency, high-purity capture and release of fetal cells, and to enable fetal genome-wide analysis

Method used

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  • Fetal cell capturing module, application method thereof, micro-fluidic chip for fetal cell capturing and application method of micro-fluidic chip for fetal cell capturing
  • Fetal cell capturing module, application method thereof, micro-fluidic chip for fetal cell capturing and application method of micro-fluidic chip for fetal cell capturing
  • Fetal cell capturing module, application method thereof, micro-fluidic chip for fetal cell capturing and application method of micro-fluidic chip for fetal cell capturing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0112] Example 1 Experimental Example of Magnetic Beads + Disulfide Bond Modified Antibody

[0113] 1.1 Experimental method Magnetic beads are selected from Thermo Fisher Company, Dynabeads TM MyOne TM CarboxylicAcid, the product number is 65012, refer to the instruction manual for the method of use, the specific display is as follows: after taking out the magnetic beads, shake and mix well, take 20 μL of magnetic beads and wash them three times with 15mM MES buffer pH 6.0, add 1-(3-dimethylaminopropyl) - 3-Ethylcarbodiimide hydrochloride solution (see instructions for concentration), react for 30 minutes, magnetically separate, wash three times with 15mM MES buffer pH 6.0, add 3-mercapto-2-propanamine, react for 30 minutes, magnetically separate, Wash with 15mM MES buffer pH 6.0 three times, then add 0.01% (mass fraction) of m-pyridyl disulfide polyethylene glycol succinimide valerate (polyethylene glycol molecular weight is 2000) solution, react for 30 minutes, Magnetic se...

Embodiment 2

[0118] Example 2 Preparation of microfluidic chip for fetal cell capture

[0119] see figure 1 structure to make a microfluidic chip. The mask plate of the chip is a silicon-based chip containing su-8 photoresist channels obtained by ultraviolet lithography. Dimethylsiloxane (PDMS) prepolymer is poured into the chip, and after pumping, heating, and degassing The PDMS channel layer containing microfluidic channels can be obtained by four steps of molding and drilling. The PDMS channel layer and the glass slide were bonded into a complete chip using a plasma cleaner (Harrick, model: PDC-002). The slide glass is preferably a sailboat brand 25.4×76.2 mm glass slide without frosted edges.

[0120] The chip is provided with two sample inlets: inlet (1) and inlet (2) and one sample outlet: outlet (3). Between the sample inlet and the sample outlet is a fluid microchannel filled with microarrays. Such as figure 1 shown. In this example, the sample inlet and the sample outlet are...

Embodiment 3

[0128] Example 3 Detecting the Capture Efficiency of Microfluidic Chips to Fetal Cells

[0129] 3.1 Experimental method

[0130] Take the microfluidic chip obtained by the recognition molecule coupling method 2 in Example 2, and use 3% bovine serum albumin solution as the blocking solution to block the chip for 30 minutes.

[0131] Add cultureable cell lines to 1mL healthy human peripheral blood to simulate the peripheral blood environment of pregnant women, (the cultureable fetal cell line is JEG-3 (human choriocarcinoma cell line), which was purchased from the Shanghai Institute of Biological Sciences, Chinese Academy of Sciences Cell Resource Center, Cat. No. TCHu195). Specifically: the number of JEG-3 cells is about 100, and the number of white blood cells is 9.63×10 6 cells / mL, red blood cells 3.98×10 9 samples / mL, the obtained blood was passed through the injection port (1) through the syringe pump, and at the same time, the buffer solution was passed through the inje...

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Abstract

The invention relates to a fetal cell capturing module, an application method thereof, a micro-fluidic chip for fetal cell capturing and an application method of the micro-fluidic chip for fetal cellcapturing. The fetal cell capturing module comprises a cell capturing carrier and a distinguishing molecule for specific capturing of cells, and the distinguishing molecule is connected to the surfaceof the carrier through an organic coupler containing disulfide bonds. The surface of the micro-fluidic chip for fetal cell capturing is modified with the distinguishing molecule used for specific capturing of the fetal cells by means of the organic coupler containing disulfide bonds, and after the distinguishing molecule captures the cells, chemical chopping is performed on the disulfide bonds inthe organic coupler to realize the release of the cells. The capturing module, the application method thereof, the micro-fluidic chip and the application method of the micro-fluidic chip can realizefetal cell capturing of whole blood without being pretreated, the capturing rate is high, the cell loss is low, the cells are simple in release operation and accurate in action, so that the fetal cells can be efficiently and harmlessly released, and complete genomes can be analyzed.

Description

technical field [0001] This application relates to the field of cell capture. It specifically relates to a fetal cell capture module, a microfluidic chip for fetal cell capture, and their use methods. Background technique [0002] Improving reproductive health and preventing and controlling major birth defects is one of the important goals of Healthy China. The "Report on the Prevention and Control of Birth Defects in China" issued by the Ministry of Health in 2012 pointed out that the total incidence of birth defects in my country is about 5.6%, and the number of new birth defects is about 900,000 every year. After the implementation of the "universal two-child" policy, the number of elderly pregnant women has increased, and the prevention and control of birth defects is also facing more challenges. "Prenatal screening and diagnosis" is the most important means of preventing birth defects. Amniocentesis, chorionic villus biopsy, and umbilical cord blood aspiration, as th...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12N5/073C12N5/078
CPCC12M23/16C12N5/0641C12N5/0605C12N5/0634B01L3/502753B01L3/502761B01L2200/0652B01L2300/0654B01L2300/0819C12N2509/00
Inventor 杨朝勇张惠敏杨园园刘艺龙朱志
Owner WISDOM HEALTHY BIOTECH XIAMEN CO LTD
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