2D and 3D cell co-culture system capable of implementing continuous harvesting without enzyme digestion and construction method and application thereof
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A technology of co-culture system and enzymatic digestion, applied in 3D culture, artificial cell constructs, cell culture support/coating, etc., can solve problems such as inability to fix, affect fixed point, timing observation, technical complexity, etc., to reduce the impact Effect
Pending Publication Date: 2021-01-12
UNIV OF ELECTRONICS SCI & TECH OF CHINA
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Problems solved by technology
However, there are still some problems to be solved in cell culture technology
[0003] 1. 2D culture conditions cannot reflect the real 3D culture environment of cells
[0004] 2. The currently commonly used 3D cell culture system needs to add collagen, sodium alginate and other gel or solid components as scaffolds. If the choice is not good, the composition of the material is very different from the real cell environment
[0005] 3. There are also some specific cells (such as tumor cells), which can form microspheres in suspension culture due to their strong self-renewal ability, but the microspheres are freely suspended in the culture medium and cannot be fixed, which affects fixed-point and regular observation.
[0006] 4. At present, compared with 2D culture, 3D cell culture has complicated technology and needs to be prepared on demand each time, which cannot achieve sustainable acquisition
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Embodiment 1
[0037] A kind of continuous harvesting of human cervical cancer cells (Hela) without enzyme digestion cell 2D, 3D co-culture system construction, comprising the following steps:
[0038] 1. Preparation of micropattern array stamps;
[0039] A rectangular pattern array with a size of 100×100 μm was prepared by UV lithography, and a PDMS stamp was prepared by using Dow Corning 184, which was cleaned for later use.
[0040] 2. Preparation of culture substrate with microarray pattern;
[0041] (1) Drop 200 μl of Fibronectin working solution onto the surface of the PDMS stamp and incubate for 30 minutes.
[0042] (2) Absorb the excess Fibronectin working solution, and dry the PDMS in an oven at 50°C.
[0043] (3) Put the PDMS stamp on a Petri dish (Guangzhou Jiete 3.5mm 2 Petri dish) on the bottom surface, apply a pressure of about 50N, and remove the PDMS stamp after 5min.
[0044] (4) Use the amphiphilic copolymer to block the sites without extracellular matrix patterns on th...
Embodiment 2
[0050] A kind of continuous harvesting of human breast cancer cells (MDA-MB-231), construction of 2D and 3D co-culture system of cells without enzyme digestion, comprising the following steps:
[0051] Diversity is the characteristic of tumor cells, each tumor cell has its own unique gene expression, protein level and biological behavior characteristics under specific conditions. The polymorphism of tumors can be better studied by taking individual cells as the research object.
[0052]1. Preparation of micropattern array stamps;
[0053] A rectangular pattern array with a size of 100×100 μm was prepared by UV lithography, and a PDMS stamp was prepared by using Dow Corning 184, which was cleaned for later use.
[0054] 2. Preparation of culture substrate with microarray pattern;
[0055] (1) Drop 200 μl of Fibronectin working solution onto the surface of the PDMS stamp and incubate for 30 minutes.
[0056] (2) Absorb the excess Fibronectin working solution, and dry the PDMS...
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Abstract
The invention discloses a 2D and 3D cell co-culture system capable of implementing continuous harvesting without enzyme digestion and a construction method and application thereof. The construction method of the 2D and 3D cell co-culture system comprises: (1) preparing a seal with a microarray pattern; (2) preparing a culture substrate with a microarray pattern; (3) co-culturing 2D and 3D cells onthe micro-pattern culture substrate; and (4) obtaining a 3D cell microsphere and 2D cell co-culture system. According to the cell 2D and 3D co-culture system capable of implementing continuous harvesting without enzyme digestion, which is provided by the invention, 2D and 3D coexsiting co-culture of the cells can be implemented in the same environment, and if 3D cell microspheres need to be obtained, the 3D cell microspheres can be harvested by simple blowing and beating through a simple suction pipe without enzyme digestion; In addition, the substrate cells can still continue to generate the3D cell microspheres just like that continuous harvesting can be implemented by planting plants, meanwhile, collagen, sodium alginate and other materials do not need to be added in the formation of the 3D cell microspheres, so as to reduce the influence of exogenous extracellular matrixes in research.
Description
technical field [0001] The invention belongs to the technical field of cell co-culture systems, and in particular relates to a 2D and 3D cell co-culture system that can be harvested continuously and does not require enzyme digestion, and its construction method and application. Background technique [0002] Cell culture is an important experimental technique in biological and medical research, and plays an irreplaceable role in basic research, drug evaluation and other experiments. However, there are still some problems to be solved urgently in cell culture technology. [0003] 1. 2D culture conditions cannot reflect the real 3D culture environment of cells. [0004] 2. The currently commonly used 3D cell culture system needs to add gel or solid components such as collagen and sodium alginate as a scaffold. If the choice is not good, the material composition will be very different from the real cell environment. [0005] 3. There are also some specific cells (such as tumor...
Claims
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Application Information
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