Microfluidic chip for separating neuroblastoma CTC and capture method thereof

A technology of neuroblastoma and microfluidic chip, which is applied in the field of microfluidic chip for separating circulating tumor cells (CTC), which can solve the problems of poor and intractable prognosis, and achieve the goal of reducing detection cost, reducing operation steps and high activity Effect

Pending Publication Date: 2021-03-12
SHENZHEN CHILDRENS HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The biological characteristics and clinical manifestations of different ages, tumor locations and different tissue differentiation levels are very

Method used

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  • Microfluidic chip for separating neuroblastoma CTC and capture method thereof
  • Microfluidic chip for separating neuroblastoma CTC and capture method thereof
  • Microfluidic chip for separating neuroblastoma CTC and capture method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Using photolithography, a microfluidic chip template capable of capturing neuroblastoma CTCs was fabricated. A polydimethylsiloxane (PDMS) prepolymer was poured on the template and cured at 80°C for 0.5h. Then the PDMS microfluidic chip is detached from the template, and holes are drilled to form blood inlet holes and waste blood outlet holes, which are bonded to the glass substrate after oxygen plasma activation treatment. The surface of the glass substrate was modified with 3-mercaptopropyltrimethoxysilane (modified with 4% v / v ethanol for 30 min at room temperature), followed by N-maleimidobutyryloxysuccinimide After ester (GMBS, 0.1 mM dimethyl sulfoxide (DMSO)) treatment for 30 min, the substrate surface was spiked with streptavidin (10 μg / ml PBS pre-treated for 1 h). Finally, the modified substrate was combined with GD2 antibody (10 μg / mL of PBS with 1.0% (w / v) BSA and 0.1% (w / v) sodium azide for 30 min) before use.

[0035] Microfluidic chips such as figure 1 ...

Embodiment 2

[0040] In order to test how the flow rate affects the capture rate, blood samples were added with fluorescently labeled neuroblastoma SK-N-SH cell line at a concentration of about 200 cells / ml, at 6, 9, 12, 15 and 18ml / The flow rate of h passes to figure 1 microfluidic chip, and then use a fluorescence microscope to observe the number of captured tumor cells. The capture efficiency is calculated according to the following formula: capture rate = N C / N S ×100%, where N C is the number of cancer cells captured, N S is the total number of cancer cells injected into the blood, and the capture efficiency represents the ratio of the number of captured tumor cells to the total number of tumor cells. Test results such as Figure 4 As shown, as the flow rate increased from 6 mL / h to 18 mL / h, the capture rate of tumor cells remained above 90%.

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Abstract

The invention discloses a microfluidic chip for separating neuroblastoma (CTC) and a capture method thereof, relates to a microfluidic chip for separating CTC, and aims to enable CTC cells to be quickly enriched and captured in the chip. The method provided by the invention realizes one-step separation and capture of the neuroblastoma CTC from whole blood based on the critical sorting size of fluid mechanics in physics and the antigen-antibody affinity effect in biochemistry, reduces the operation steps, and improves the capture efficiency and cell activity of the neuroblastoma CTC. The methodhas the advantages that one-step operation is achieved, and complex pretreatment is not needed; 2, the capture efficiency and purity of CTC are improved through two-stage separation; 3, the CTC cellswith high efficiency and high activity are finally obtained under the dual capture action; 4, the cells can be put under the microscope for observing and detecting.

Description

technical field [0001] The invention relates to a microfluidic chip for isolating circulating tumor cells (CTCs), in particular to a microfluidic chip for isolating neuroblastoma CTCs and a capture method thereof. Background technique [0002] Neuroblastoma (NB) is the most common extracranial solid tumor in infants and young children, accounting for 8% to 10% of childhood malignancies. Neuroblastoma is a group of diseases with very different clinical manifestations and prognosis, ranging from tumor dissemination, metastasis, death of children, to tumor maturation into benign ganglioneuroma or spontaneous regression and other clinical outcomes. The adrenal gland is the most common primary site, followed by the abdominal sympathetic ganglion, thoracic sympathetic ganglion, cervical sympathetic ganglion, and pelvic sympathetic ganglion. About 1% of patients failed to find the primary tumor. Neuroblastoma can metastasize to lymph nodes, bone marrow, bone, dura mater, orbit, li...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12N5/09B01L3/00
CPCC12M47/04C12N5/0693C12N5/0618B01L3/5027B01L3/502707C12N2509/00
Inventor 袁秀丽陈小文文飞球宋建明刘宗彬
Owner SHENZHEN CHILDRENS HOSPITAL
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