CN103060175A - Cell micro-array chip and preparation method thereof - Google Patents
Cell micro-array chip and preparation method thereof Download PDFInfo
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- CN103060175A CN103060175A CN2013100016626A CN201310001662A CN103060175A CN 103060175 A CN103060175 A CN 103060175A CN 2013100016626 A CN2013100016626 A CN 2013100016626A CN 201310001662 A CN201310001662 A CN 201310001662A CN 103060175 A CN103060175 A CN 103060175A
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Abstract
The invention provides a cell micro-array chip and a preparation method thereof and belongs to the technical field of cell chips. The cell micro-array chip is characterized by being prepared by using alginate gels; and the preparation method comprises the following steps: coating a layer of sodium alginate solution on a glass plate, rapidly immerging the glass plate in a calcium chloride solution and soaking the glass plate for 5-30 minutes to form a gel template; preparing a dot matrix template on which a plurality of cylinders are arranged, soaking the top ends of the cylinders in a chitosan or polylysine solution and taking out the cylinders after 5-30 minutes; embossing the top ends of the cylinders on the alginate gel template and forming a complex dot matrix template through electrostatic complexation reaction; soaking the complex dot matrix template in a culture solution with cell concentration of 1*105-1*107 and forming the alginate gel cell micro-array chip when the complex dot matrix template is cultured for 4-16 hours; and the cell micro-array chip provided by the invention has the advantage of simplicity and convenience in operation and can be used for cell tissue like culturing, high throughput screening of compounds or medicines, discovery of new genes, proteomic studies and the like.
Description
Technical field:
The invention belongs to the cell chip technical field, be specifically related to a kind of cell microarray chip and preparation method thereof.
Background technology:
Biochip mainly is that biologically active substance is assembled on the solid substrate, consists of microarray, to realize screening or the detection to accurate, the quick and bulk information of compound (such as medicine), protein, cell and other biological component.Cell microarray chip is a kind of of biochip, namely by micro-processing technology, makes the check-out console that density is higher, the aperture is less at substrates such as silicon or glass or plastics, and cell is fixed in the hole.The maturation of this technology has extremely important meaning for the detection of exploitation, food, environment and the life science of new drug.
At present, existing technology mainly with silicon or glass or plastics etc. as base material, the method for distributing by micro-sampling or other trace, with the surface of cell assembling to base material, prepare cell microarray chip, complex manufacturing process is not easy to simultaneously operate continuously.
Summary of the invention:
The object of the present invention is to provide a kind of cell microarray chip and preparation method thereof, can effectively overcome the shortcoming that prior art exists.
The present invention is achieved in that the cell microarray chip that it is characterized in that by the preparation of Lalgine gel, and as shown in Figure 1, its preparation method is:
The first step, preparation alginic acid hydrogel template: the mass concentration that is about to preparation is that the sodium alginate soln of 0.8～4.0 g/L is coated in above the sheet glass, then sheet glass is immersed in fast in the calcium chloride solution that mass concentration is 5～30 g/L, soak after 5～30 minutes and take out, can form one deck Lalgine gel at sheet glass, be called alginic acid hydrogel template 1.
Second step, preparation polydimethylsiloxane dot matrix template: namely prepare one and be arranged with in the above a plurality of cylindrical lattice mode templates, right cylinder 2 tops on this mould plate are immersed in the chitosan solution or polylysine solution that mass concentration is 0.05～3.0 g/L, soaked 5～30 minutes, take out, make polydimethylsiloxane dot matrix template 3.
The 3rd step, right cylinder 2 tops above the polydimethylsiloxane dot matrix template 3 are stamped on the alginic acid hydrogel template 1, reacted 5～20 minutes, chitosan or polylysine can be infiltrated in the Lalgine gel, form complex compound lattice point 4 by the static complex reaction, and form complex compound dot matrix template 5.
In the 4th step, inoculating cell: it is 1 * 10 that the Lalgine gel template 5 that will process is immersed in cell concn
5～1 * 10
7Nutrient solution in, cultivated 4～16 hours, cell 6 self aggregations also adhere on the dot matrix, form cell microarray chip 7.
The method is easy and simple to handle, can be used for the research of the discovery of the high flux screening of class Organoid culture, compound or the medicine of cell, new gene and gene function and proteomics research etc.
Description of drawings
Fig. 1 is that cell chip of the present invention prepares synoptic diagram
1-alginic acid hydrogel template;
2-right cylinder;
3-polydimethylsiloxane template;
4-complex compound lattice point;
5-complex compound dot matrix template;
6-cell;
7-cell microarray chip.
Embodiment
Embodiment 1: at the micro-array chip of Lalgine gel preparation HepG2 cell
The HepG2 cell is a kind of of hepatoma cell line.
The first step, preparation alginic acid hydrogel template: the mass concentration that is about to preparation is that the sodium alginate soln of 1.0 g/L is coated in above the sheet glass, then sheet glass is immersed in fast in the calcium chloride solution that mass concentration is 10 g/L, soak after 30 minutes and take out, can form one deck Lalgine gel at sheet glass, be called the alginic acid hydrogel template.
Second step, preparation polydimethylsiloxane dot matrix template: the lattice mode board size is 30 * 30 mm, be arranged with a plurality of right cylinders above it, cylindrical cross-sectional dimension is 200 μ m, distance between each right cylinder center is 600 μ m, then and with the top of this template be dipped in the chitosan solution solution that mass concentration is 0.5 g/L, soaked 30 minutes.
The 3rd step was stamped in the right cylinder top above the polydimethylsiloxane dot matrix template on the alginic acid hydrogel template, reacted 20 minutes, formed static complex compound dot matrix template.
In the 4th step, inoculating cell: it is 1 * 10 that the Lalgine gel template 3 that will process is immersed in cell concn
5Nutrient solution in, cultivated 16 hours, the cell self aggregation also adheres on the dot matrix, forms the HepG2 cell microarray chip.
Embodiment 2: at the micro-array chip of Lalgine gel preparation Chinese hamster ovary celI
Chinese hamster ovary celI is a clone of Chinese hamster ovary cell.
The first step, preparation alginic acid hydrogel template: the mass concentration that is about to preparation is that the sodium alginate soln of 2.0 g/L is coated in above the sheet glass, then sheet glass is immersed in fast in the calcium chloride solution that mass concentration is 15 g/L, soak after 20 minutes and take out, can form one deck Lalgine gel at sheet glass, be called the alginic acid hydrogel template.
Second step, preparation polydimethylsiloxane dot matrix template: the lattice mode board size is 30 * 30 mm, be arranged with a plurality of right cylinders above it, cylindrical cross-sectional dimension is 200 μ m, distance between each right cylinder center is 600 μ m, then and with the top of this template be dipped in the chitosan solution solution that mass concentration is 3.0 g/L, soaked 5 minutes.
The 3rd step was stamped in the right cylinder top above the polydimethylsiloxane dot matrix template on the alginic acid hydrogel template, reacted 15 minutes.
In the 4th step, inoculating cell: it is 1 * 10 that the Lalgine gel template that will process is immersed in cell concn
7Nutrient solution in, cultivated 4 hours, the cell self aggregation also adheres on the dot matrix, forms the Chinese hamster ovary celI micro-array chip.
Embodiment 3: at the micro-array chip of Lalgine gel preparation McF7 cell
The McF7 cell is a kind of of breast cancer cell line.
The first step, preparation alginic acid hydrogel template: the mass concentration that is about to preparation is that the sodium alginate soln of 4.0 g/L is coated in above the sheet glass, then sheet glass is immersed in fast in the calcium chloride solution that mass concentration is 30 g/L, soak after 5 minutes and take out, can form one deck Lalgine gel at sheet glass, be called the alginic acid hydrogel template.
Second step, preparation polydimethylsiloxane dot matrix template: the lattice mode board size is 30 * 30 mm, be arranged with a plurality of right cylinders above it, cylindrical cross-sectional dimension is 200 μ m, distance between each right cylinder center is 600 μ m, then and with the top of this template be dipped in the polylysine solution solution that mass concentration is 0.05 g/L, soaked 30 minutes.
The 3rd step was stamped in the right cylinder top above the polydimethylsiloxane dot matrix template on the alginic acid hydrogel template, reacted 5 minutes.
In the 4th step, inoculating cell: it is 2 * 10 that the Lalgine gel template that will process is immersed in cell concn
6Nutrient solution in, cultivated 8 hours, the cell self aggregation also adheres on the dot matrix, forms the McF7 cell microarray chip.
Claims (2)
1. cell microarray chip is characterized in that by the preparation of Lalgine gel.
2. the cell microarray chip by Lalgine gel preparation as claimed in claim 1 is characterized in that the preparation method is:
The first step, preparation alginic acid hydrogel template: the mass concentration that is about to preparation is that the sodium alginate soln of 0.8～4.0 g/L is coated in above the sheet glass, then sheet glass is immersed in fast in the calcium chloride solution that mass concentration is 5～30 g/L, soak after 5～30 minutes and take out, can form one deck Lalgine gel at sheet glass, be called alginic acid hydrogel template (1);
Second step, preparation polydimethylsiloxane dot matrix template: namely prepare a lattice mode template that is arranged with in the above a plurality of right cylinders (2), right cylinder (2) top on this mould plate is immersed in the chitosan solution or polylysine solution that mass concentration is 0.05～3.0 g/L, soaked 5～30 minutes, take out, make polydimethylsiloxane dot matrix template (3);
The 3rd step, right cylinder top above the polydimethylsiloxane dot matrix template (3) is stamped on the alginic acid hydrogel template (1), reacted 5～20 minutes, chitosan or polylysine can be infiltrated in the Lalgine gel, generate complex compound lattice point (4) by the static complex reaction, and form complex compound dot matrix template (5);
In the 4th step, inoculating cell: it is 1 * 10 that complex compound dot matrix template (5) is immersed in cell concn
5～1 * 10
7Nutrient solution in, cultivated 4～16 hours, cell (6) self aggregation also adheres on the dot matrix, forms the cell microarray chip (7) by the preparation of Lalgine gel.
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Cited By (7)
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| CN103351484A (en) * | 2013-07-03 | 2013-10-16 | 苏州睿研纳米医学科技有限公司 | Micropatterned hydrogel coating, its preparation method and use |
| CN103387206A (en) * | 2013-07-18 | 2013-11-13 | 中国科学院合肥物质科学研究院 | Manufacturing method of ultramicropore broadband flexible micro-perforated plate |
| CN105950467A (en) * | 2016-05-18 | 2016-09-21 | 清华大学 | Method for detecting cytotoxicity of to-be-tested medicine to target cell and cell chip specially used by method |
| CN106238111A (en) * | 2016-07-28 | 2016-12-21 | 南京理工大学 | A kind of microcapsule preparation method based on micro-fluidic chip shear flow |
| CN109312300A (en) * | 2016-04-06 | 2019-02-05 | 微电子联合有限公司 | Sphere micro-array tissue and preparation method |
| CN110452801A (en) * | 2019-07-04 | 2019-11-15 | 中国科学技术大学 | A kind of micro-fluidic chip and preparation method thereof and catching method |
| CN113138249A (en) * | 2021-04-12 | 2021-07-20 | 北京蛋白质组研究中心 | Micro-sample metabolome, proteome and phosphoproteome multi-group chemical analysis method based on micropore array chip |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103351484A (en) * | 2013-07-03 | 2013-10-16 | 苏州睿研纳米医学科技有限公司 | Micropatterned hydrogel coating, its preparation method and use |
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| CN109312300A (en) * | 2016-04-06 | 2019-02-05 | 微电子联合有限公司 | Sphere micro-array tissue and preparation method |
| JP2019513419A (en) * | 2016-04-06 | 2019-05-30 | マイクロマトリシーズ アソシエイツ リミテッド | Spheroid tissue microarray and method for producing the same |
| CN105950467A (en) * | 2016-05-18 | 2016-09-21 | 清华大学 | Method for detecting cytotoxicity of to-be-tested medicine to target cell and cell chip specially used by method |
| CN105950467B (en) * | 2016-05-18 | 2018-08-28 | 清华大学 | A kind of method detecting drug to be measured to the cytotoxicity of aim cell and its special cell chip |
| CN106238111A (en) * | 2016-07-28 | 2016-12-21 | 南京理工大学 | A kind of microcapsule preparation method based on micro-fluidic chip shear flow |
| CN110452801A (en) * | 2019-07-04 | 2019-11-15 | 中国科学技术大学 | A kind of micro-fluidic chip and preparation method thereof and catching method |
| CN113138249A (en) * | 2021-04-12 | 2021-07-20 | 北京蛋白质组研究中心 | Micro-sample metabolome, proteome and phosphoproteome multi-group chemical analysis method based on micropore array chip |
| CN113138249B (en) * | 2021-04-12 | 2021-11-23 | 北京蛋白质组研究中心 | Micro-sample metabolome, proteome and phosphoproteome multi-group chemical analysis method based on micropore array chip |
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