Array chip for cell capture and tumor ball culture and preparation and operation method thereof

A cell culture and tumor sphere technology, applied in the field of array chip preparation, can solve the problems of single cell chip function and single cell waste, and achieve the effects of simple and fast operation, simple and easy preparation method and operation method.

Pending Publication Date: 2019-10-08
XIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide an arrayed chip for cell capture and tumor sphere culture, by designing a complex microfiltration structure to achieve the capture of single cells of different sizes and deformability, and then by combining with the microporous structure to achieve The long-term culture and subsequent detection of captured single cells solves the problem that the cell chip with the existing microfiltration structure has a single function and cannot be cultured in the later stage, resulting in the waste of captured single cells

Method used

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  • Array chip for cell capture and tumor ball culture and preparation and operation method thereof
  • Array chip for cell capture and tumor ball culture and preparation and operation method thereof
  • Array chip for cell capture and tumor ball culture and preparation and operation method thereof

Examples

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preparation example Construction

[0051] The method for preparing an arrayed chip for cell capture and tumorsphere culture of the present invention specifically includes the following steps:

[0052] Step 1, prepare single cell sorting layer:

[0053] Mix the PDMS matrix and curing agent at a mass ratio of 5:1. At the same time, use trimethylchlorosilane (Trimethylchlorosilan, TMCS) steam to treat the single-cell separation layer mold for 5 minutes to 10 minutes, and pour the mixture of PDMS matrix and curing agent into the trimethyl Put the chlorosilane-treated single-cell sorting layer mold on the vacuum degassing and place it in an oven at 80°C to 100°C for heating and curing for 0.5h to 1h, peel the cured PDMS from the mold, and cut it as required. And punch holes to prepare screening tanks, clean them for future use, wherein, the PDMS matrix and curing agent are purchased from Dow Corning Corporation of the United States, number: SYLGARD 184.

[0054] Step 2, prepare cell culture layer:

[0055] Mix the...

Embodiment 1

[0064] The applicant's laboratory of the present invention designs the structure and size of the micro-control chip, such as figure 2 As shown, A and B represent the width and length of the microstructure in the single-cell sorting layer, respectively, and the values ​​are 120 μm and 200 μm, respectively, and the width of the constriction in the middle is 90 μm. C represents the distance between the microarrays, and the value is 50 μm, D and E represent the size of the first pore and the second pore between two adjacent microstructures are the same in the capture units of the same column, but their sizes are sequentially reduced by 2 μm in the capture units of different microarrays (ie The size of the first pore in this example is 16 to 8 μm, and the size of the second pore is 14 to 6 μm), such as image 3 As shown, the length of the screening groove formed between two adjacent microstructures is 100 μm, the width of the widest part in the middle is 40 μm, the height L1 of th...

Embodiment 2

[0079] The applicant's laboratory of the present invention designs the structure and size of the micro-control chip, such as figure 2 As shown, A and B represent the width and length of the microstructure in the single-cell sorting layer, respectively, the values ​​are 120 μm and 250 μm, and the width of the constriction in the middle is 90 μm, C represents the distance between the microarrays, the value is 50 μm, D and E represent the size of the first pore and the second pore between two adjacent microstructures are the same in the capture units of the same column, but their sizes are sequentially reduced by 2 μm in the capture units of different microarrays (ie The size of the first pore in this example is 16 to 8 μm, and the size of the second pore is 14 to 6 μm), such as image 3 As shown, the length of the screening groove formed between two adjacent microstructures is 120 μm, the width of the widest part in the middle is 50 μm, the height L1 of the screening groove of ...

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Abstract

The invention relates to an array chip for cell capture and tumor ball culture, which comprises a single cell sorting layer and a cell culture layer, and a cell culture layer is adhered to the singlecell sorting layer. The single cell sorting layer includes at least one set of cell capture arrays, a cell suspension liquid inlet, and a cell suspension liquid outlet, each set of cell capture arrayscomprises at least three columns of microarrays, each column of microarrays includes single cell capture units arranged side by side, and the single cell capture units comprise two microstructures, ascreening groove is formed between the two microstructures, a first pore and a second pore for capturing the cells are respectively formed at two ends of the screening groove, and a width of the first pore is larger than the width of the second pore by 2 [mu]m; the cell culture layer is provided with a culture tank, and the culture tank corresponds to the single cell capture unit. The invention also discloses a preparation method and an operation method of the chip, which have the characteristics of simple and quick operation and low consumption, and can be widely used in a variety of parallel high-throughput and multi-repetition single-cell operation and analysis applications.

Description

technical field [0001] The invention belongs to the field of cell biology and microfluidic chip technology, and specifically relates to an arrayed chip for cell capture and tumor sphere culture, and also relates to a preparation method for an array chip for cell capture and tumor sphere culture, and also relates to a chip array for cell capture and tumor sphere culture. An operation method of an arrayed chip for cell capture and tumorsphere culture. Background technique [0002] At present, research and analysis based on single-cell level is widely used in biological research and clinical detection, and has grown into a new analysis method and detection platform. Microfluidic chip technology is a micro-operation and analysis method that has just emerged in this century. It has shown great potential for single-cell manipulation and analysis since its inception. So far, single cell capture and manipulation methods based on microfluidic chips mainly include: microstructure fi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/12C12M1/00
CPCC12M33/14C12M23/02
Inventor 庞龙葛玉鑫袁皓月范江霖范士冈郭陆露靳娅茹陈镝
Owner XIAN MEDICAL UNIV
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