CN102276866B - Plasma-photochemical method for grafting carboxyl on cell culture surface - Google Patents
Plasma-photochemical method for grafting carboxyl on cell culture surface Download PDFInfo
- Publication number
- CN102276866B CN102276866B CN201110200304A CN201110200304A CN102276866B CN 102276866 B CN102276866 B CN 102276866B CN 201110200304 A CN201110200304 A CN 201110200304A CN 201110200304 A CN201110200304 A CN 201110200304A CN 102276866 B CN102276866 B CN 102276866B
- Authority
- CN
- China
- Prior art keywords
- cell culturing
- plasma
- culturing surfaces
- solution
- cultivation surface
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 38
- 238000004113 cell culture Methods 0.000 title abstract description 13
- 239000007788 liquid Substances 0.000 claims abstract description 38
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000000178 monomer Substances 0.000 claims abstract description 20
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims abstract description 16
- 229910052760 oxygen Inorganic materials 0.000 claims abstract description 15
- 239000001301 oxygen Substances 0.000 claims abstract description 15
- 238000012258 culturing Methods 0.000 claims description 65
- 239000000463 material Substances 0.000 claims description 21
- -1 polyethylene terephthalate Polymers 0.000 claims description 17
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 claims description 15
- 238000002203 pretreatment Methods 0.000 claims description 13
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 claims description 12
- 230000015572 biosynthetic process Effects 0.000 claims description 12
- 239000007789 gas Substances 0.000 claims description 12
- NIXOWILDQLNWCW-UHFFFAOYSA-N Acrylic acid Chemical compound OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims description 11
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 claims description 11
- CCJAYIGMMRQRAO-UHFFFAOYSA-N 2-[4-[(2-hydroxyphenyl)methylideneamino]butyliminomethyl]phenol Chemical compound OC1=CC=CC=C1C=NCCCCN=CC1=CC=CC=C1O CCJAYIGMMRQRAO-UHFFFAOYSA-N 0.000 claims description 10
- 125000004494 ethyl ester group Chemical group 0.000 claims description 10
- 229920000139 polyethylene terephthalate Polymers 0.000 claims description 6
- 239000005020 polyethylene terephthalate Substances 0.000 claims description 6
- 229930040373 Paraformaldehyde Natural products 0.000 claims description 5
- 229920003229 poly(methyl methacrylate) Polymers 0.000 claims description 5
- 229920000515 polycarbonate Polymers 0.000 claims description 5
- 239000004417 polycarbonate Substances 0.000 claims description 5
- 239000004926 polymethyl methacrylate Substances 0.000 claims description 5
- 229920006324 polyoxymethylene Polymers 0.000 claims description 5
- 239000004698 Polyethylene Substances 0.000 claims description 4
- 229920000573 polyethylene Polymers 0.000 claims description 4
- FPYJFEHAWHCUMM-UHFFFAOYSA-N maleic anhydride Chemical compound O=C1OC(=O)C=C1 FPYJFEHAWHCUMM-UHFFFAOYSA-N 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 210000000130 stem cell Anatomy 0.000 abstract description 14
- 239000003960 organic solvent Substances 0.000 abstract description 10
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 230000001678 irradiating effect Effects 0.000 abstract description 2
- 239000003999 initiator Substances 0.000 abstract 2
- 239000002344 surface layer Substances 0.000 abstract 2
- 238000004140 cleaning Methods 0.000 abstract 1
- 238000007781 pre-processing Methods 0.000 abstract 1
- 238000003892 spreading Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 55
- 239000000243 solution Substances 0.000 description 44
- 239000004793 Polystyrene Substances 0.000 description 17
- 238000005516 engineering process Methods 0.000 description 10
- 230000001464 adherent effect Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 238000009736 wetting Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 125000003636 chemical group Chemical group 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012776 electronic material Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 210000001178 neural stem cell Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000009965 odorless effect Effects 0.000 description 1
- 238000011197 physicochemical method Methods 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000009168 stem cell therapy Methods 0.000 description 1
- 238000009580 stem-cell therapy Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000013077 target material Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229920001169 thermoplastic Polymers 0.000 description 1
- 239000004416 thermosoftening plastic Substances 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Treatments Of Macromolecular Shaped Articles (AREA)
Abstract
The invention discloses a plasma-photochemical method for grafting carboxyl on a cell culture surface. The method comprises the following steps of: preprocessing the cell culture surface in a plasma reaction chamber, and exposing in air or oxygen for 25 minutes to 24 hours; dissolving a monomer in water to form solution A; spreading the solution A to culture surface obtained by the step (2) uniformly to form a liquid surface layer; irradiating the culture surface on which the liquid surface layer is spread by ultraviolet (UV); and taking the culture surface out of the solution A, and cleaning by the water to obtain the cell culture surface on which the carboxyl is grafted. By the plasma-photochemical method, the carboxyl content of the cell culture surface is improved effectively without using an organic solvent and a light initiator; the damage of the organic solvent and the light initiator to the environment and human bodies in the production process and the potential impact of subsequent stem cell culture are avoided successfully; a step of removing residues is simplified, the production period is shortened, and the cost is saved.
Description
Technical field
The invention belongs to the material engineering field, be specifically related to a kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting.
Background technology
Along with the continuous development of modern science, especially biomedical progressive fast, stem cell is cultivated becomes crucial basic work in a lot of researchs and the Application Areas.All the time, people generally adopt culture plate or the culturing bottle of the PS material instrument as cell cultures.PS is a kind of water white thermoplastics, is generally nontoxic, odorless, colourless transparent grain, and like glassy hard brittle material, general formula is [(CH2CHC6H5) n], and English name is Polystyrene, is called for short PS.Its goods have superior anti-water, moistureproofness.In people's productive life, the purposes of PS is a big purposes wherein as cell cultures very extensively.
The cell culturing surfaces of PS material is nonpolar, and wetting ability is very poor, is unfavorable for the adherent growth of stem cell.Therefore, people usually can carry out modification or processing to cultivation surface before culturing stem cells, and derived a new research direction---the cell culturing surfaces treatment technology in the biomedicine thus.
In research in the past and the application, cultivating the more method of employing to stem cell has: carry out biology in cultivation surface and encapsulate, encapsulate as in Culture of neural stem cells, with Laminin (ln) cultivation surface being carried out biology; Or in substratum, add the adherent performance that serum strengthens cell, like the cultivation of mescenchymal stem cell; Or cultivation surface is carried out modification through some physicochemical methods; Handle cell culturing surfaces like using plasma; The group that introducing contains elements such as oxygen, nitrogen improves its surperficial wetting ability, improves the adherent performance of stem cell on its surface through improving wetting ability.Yet; All there are some deficiencies objectively in these traditional methods: the various coating proteins (as: Laminin) as surperficial substrate capsulating material cost an arm and a leg; To limited funds in any relevant problem all is an acid test; And these albumen generally are the albumen of animal-origin, bring certain risk can for human stem-cell therapy; Serum be used for stem cell cultivate have that composition is indeterminate, the source is complicated and each batch between because problem such as individual different othernesses are bigger, this experimental study and clinical application of cultivating to stem cell has increased bigger risk; The modification of simple using plasma technology is handled, and is unsatisfactory under the serum-free culture condition.
In recent years, be accompanied by the huge advance made of Materials science and nanotechnology, the correlation technique of plasma body and photochemistry grafting functional property chemical group has been applied in the middle of the industrial goods such as electronic material, automotive material.Abroad; The science researcher handles these two kinds of technology introduction to the stem cell cultivation surface in the middle of; Through carrying out condensation reaction at PS surface grafting carboxyl and with polypeptide, polypeptide is easy to by the identification of stem cell surface receptor-specific and combines, thereby promotes the adherent growth of cell.Wherein plasma body combination photochemical method is the important step that obtains to contain a large amount of carboxyl cultivation surface, also is to realize one of this technological committed step.
But combine in the correlation technique of photochemical method carboxyl grafting at plasma body; Existing technology adopts volatile organic solvent that monomer is dissolved and dilutes usually; Then with the monomer deposition of dilution at the substrate surface of goods and evaporate organic solvent, in uv irradiating is handled, also need add light trigger simultaneously.Owing to introduced organic solvent and light trigger, made this technology exist some and still remain to be changed perfect place.At first, the intervention of organic solvent and light trigger unavoidably causes residual, and experimental study and clinical application that these residues are cultivated follow-up stem cell all will produce certain spinoff and risk.Secondly, adopt the method for organic solvent itself that environment and human body are worked the mischief easily.Once more, itself will produce these materials than high cost, adds the follow-up residual expense of removing, and the application of result that makes this technology is become very expensive.At last, existing technology is because of the characteristic of himself, and program is complicated in practice, time and effort consuming.
Summary of the invention
Shortcoming and deficiency in order to overcome prior art the object of the present invention is to provide a kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting, and this method adopts water as monomeric diluent, replace used organic solvent in the existing method; And shine through the pretreated surface of plasma body formed peroxo-layer through UV-light (UV), realize monomeric grafting, avoided in the grafting process, using light trigger thereby make it produce avtive spot.Can obtain the surface of high-carboxyl-content easily through the method for improvement.
The object of the invention is realized through following technical proposals:
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting; Be that using plasma technology pair cell cultivation surface is carried out pre-treatment; Make its surface produce radical, this surface is placed in the air or oxygen forms the peroxo-layer with the oxygen effect, the peroxo-layer produces avtive spot under UV-irradiation; Make monomer be grafted to cell culturing surfaces, thereby obtain to contain the surface of carboxyl through the chemical bond mode.Its concrete steps comprise:
(1) cell culturing surfaces is put into plasma-reaction-chamber and carry out pre-treatment;
(2) pretreated cultivation surface is placed on 25min～24h in the air or oxygen;
(3) with the monomer formation solution A that is dissolved in the water; Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the cell culturing surfaces of carboxyl grafting.
Cell culturing surfaces as herein described is meant when culturing cell, in the cell cultures vessel (like culture plate, culturing bottle, petridish etc.) with the face of cells contacting.
The material of said cell culturing surfaces can be a kind of in PS, Vilaterm, Vestolen PP 7052, polymethylmethacrylate, polyethylene terephthalate, polycarbonate, polyoxymethylene or polyethylene terephthalate-1,4 cyclohexane dimethanol ester;
The working parameter of the said plasma-reaction-chamber of step (1) is: power 35～600W, pressure 10～80Pa, gas flow rate 5～500cc/min, 20～35 ℃ of temperature;
The said pretreated time of step (1) is 30s～10min;
The said monomer of step (3) is the two or more mixtures in vinylformic acid, methylacrylic acid, Rocryl 400, maleic anhydride, N-vinyl pyrrolidone, vinylformic acid-2-carboxylic ethyl ester or the N hydroxymethyl acrylamide;
In the said solution A of step (3), the monomer total mass concentration is 11.5-15.5%;
The height of the said liquid level layer of step (3) is 0.5～2mm;
The described UV irradiation of step (4), its correlation parameter is: UV power of lamp tube 300～1000W, irradiation time 2～30min, the distance 3～13cm of UV fluorescent tube and cultivation surface.
The present invention has following advantage and effect with respect to prior art:
(1) the carboxyl grafting cell culturing surfaces for preparing of the inventive method, a large amount of carboxyls on its surface can be used as functional groups to carry out chemistry with the succeeding target material and is connected, thus the surface that realization has specific functionality.Like substrate surface and special peptides covalent attachment, can cultivate for stem cell experiment consumptive material efficient, of fine quality is provided.
(2) the inventive method does not adopt organic solvent and light trigger in the carboxyl-content that improves cell culturing surfaces effectively; Successfully avoided organic solvent and light trigger in process of production the harm of environment and human body and follow-up stem cell are cultivated in potential impact; Simplified and removed residual step, shortened the production cycle widely, also helped practicing thrift cost, thereby make this technology more help suitability for industrialized production.
Embodiment
Below in conjunction with embodiment the present invention is described in further detail, but embodiment of the present invention is not limited thereto.
Embodiment 1
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of PS material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 35W, pressure 10Pa, gas flow rate 5cc/min, 20 ℃ of temperature, treatment time 10min;
(2) pretreated cultivation surface is placed on 32min in the air or oxygen;
(3) with monomer vinylformic acid 4.0% (all refer to its mass concentration in solution A, down with), methylacrylic acid 4.0%, the Rocryl 400 3.5% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 1mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 1000W, irradiation time 2min, UV fluorescent tube and cultivation surface apart from 13cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the PS cell culturing surfaces of carboxyl grafting.
The PS cell culturing surfaces carboxyl-content that present embodiment prepares is 52.59nmol/cm
2
Embodiment 2
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of PS material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 350W, pressure 30Pa, gas flow rate 200cc/min, 28 ℃ of temperature, treatment time 150s;
(2) pretreated cultivation surface is placed on 25min in the air or oxygen;
(3) with monomer methylacrylic acid 5.0% (all refer to its mass concentration in solution A, down with), maleic anhydride 3.0%, N-vinyl pyrrolidone 2.0%, the Rocryl 400 3.0% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 0.5mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 500W, irradiation time 15min, UV fluorescent tube and cultivation surface apart from 8cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the PS cell culturing surfaces of carboxyl grafting.
The PS cell culturing surfaces carboxyl-content that present embodiment prepares is 49.16nmol/cm
2
Embodiment 3
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of PS material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 600W, pressure 80Pa, gas flow rate 500cc/min, 35 ℃ of temperature, treatment time 30s;
(2) pretreated cultivation surface is placed on 24h in the air or oxygen;
(3) with monomer vinylformic acid 4.0% (all refer to its mass concentration in solution A, down with), vinylformic acid-2-carboxylic ethyl ester 3.0%, methylacrylic acid 2.0%, N hydroxymethyl acrylamide 2.5%, the N-vinyl pyrrolidone 4.0% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 2mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 300W, irradiation time 30min, UV fluorescent tube and cultivation surface apart from 3cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the PS cell culturing surfaces of carboxyl grafting.
The PS cell culturing surfaces carboxyl-content that present embodiment prepares is 55.73nmol/cm
2
Embodiment 4
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of Vilaterm material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 600W, pressure 80Pa, gas flow rate 500cc/min, 35 ℃ of temperature, treatment time 30s;
(2) pretreated cultivation surface is placed on 24h in the air or oxygen;
(3) with monomer vinylformic acid 4.0% (all refer to its mass concentration in solution A, down with), vinylformic acid-2-carboxylic ethyl ester 3.0%, methylacrylic acid 2.0%, N hydroxymethyl acrylamide 2.5%, the N-vinyl pyrrolidone 4.0% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 2mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 300W, irradiation time 30min, UV fluorescent tube and cultivation surface apart from 3cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the Vilaterm cell culturing surfaces of carboxyl grafting.
The Vilaterm cell culturing surfaces carboxyl-content that present embodiment prepares is 37.48nmol/cm
2
Embodiment 5
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of Vestolen PP 7052 material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 600W, pressure 80Pa, gas flow rate 500cc/min, 35 ℃ of temperature, treatment time 30s;
(2) pretreated cultivation surface is placed on 24h in the air or oxygen;
(3) with monomer vinylformic acid 4.0% (all refer to its mass concentration in solution A, down with), vinylformic acid-2-carboxylic ethyl ester 3.0%, methylacrylic acid 2.0%, N hydroxymethyl acrylamide 2.5%, the N-vinyl pyrrolidone 4.0% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 2mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 300W, irradiation time 30min, UV fluorescent tube and cultivation surface apart from 3cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the Vestolen PP 7052 cell cultivation surface of carboxyl grafting.
The Vestolen PP 7052 cell cultivation surface carboxyl-content that present embodiment prepares is 54.76nmol/cm
2
Embodiment 6
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of polymethylmethacrylate material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 600W, pressure 80Pa, gas flow rate 500cc/min, 35 ℃ of temperature, treatment time 30s;
(2) pretreated cultivation surface is placed on 24h in the air or oxygen;
(3) with monomer vinylformic acid 4.0% (all refer to its mass concentration in solution A, down with), vinylformic acid-2-carboxylic ethyl ester 3.0%, methylacrylic acid 2.0%, N hydroxymethyl acrylamide 2.5%, the N-vinyl pyrrolidone 4.0% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 2mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 300W, irradiation time 30min, UV fluorescent tube and cultivation surface apart from 3cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the polymethylmethacrylate cell culturing surfaces of carboxyl grafting.
The polymethylmethacrylate cell culturing surfaces carboxyl-content that present embodiment prepares is 64.62nmol/cm
2
Embodiment 7
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of polyethylene terephthalate material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 600W, pressure 80Pa, gas flow rate 500cc/min, 35 ℃ of temperature, treatment time 30s;
(2) pretreated cultivation surface is placed on 24h in the air or oxygen;
(3) with monomer vinylformic acid 4.0% (all refer to its mass concentration in solution A, down with), vinylformic acid-2-carboxylic ethyl ester 3.0%, methylacrylic acid 2.0%, N hydroxymethyl acrylamide 2.5%, the N-vinyl pyrrolidone 4.0% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 2mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 300W, irradiation time 30min, UV fluorescent tube and cultivation surface apart from 3cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the polyethylene terephthalate cell culturing surfaces of carboxyl grafting.
The polyethylene terephthalate cell culturing surfaces carboxyl-content that present embodiment prepares is 40.53nmol/cm
2
Embodiment 8
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of polycarbonate material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 600W, pressure 80Pa, gas flow rate 500cc/min, 35 ℃ of temperature, treatment time 30s;
(2) pretreated cultivation surface is placed on 24h in the air or oxygen;
(3) with monomer vinylformic acid 4.0% (all refer to its mass concentration in solution A, down with), vinylformic acid-2-carboxylic ethyl ester 3.0%, methylacrylic acid 2.0%, N hydroxymethyl acrylamide 2.5%, the N-vinyl pyrrolidone 4.0% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 2mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 300W, irradiation time 30min, UV fluorescent tube and cultivation surface apart from 3cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the polycarbonate cell culturing surfaces of carboxyl grafting.
The polycarbonate cell culturing surfaces carboxyl-content that present embodiment prepares is 51.13nmol/cm
2
Embodiment 9
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of polyoxymethylene material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 600W, pressure 80Pa, gas flow rate 500cc/min, 35 ℃ of temperature, treatment time 30s;
(2) pretreated cultivation surface is placed on 24h in the air or oxygen;
(3) with monomer vinylformic acid 4.0% (all refer to its mass concentration in solution A, down with), vinylformic acid-2-carboxylic ethyl ester 3.0%, methylacrylic acid 2.0%, N hydroxymethyl acrylamide 2.5%, the N-vinyl pyrrolidone 4.0% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 2mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 300W, irradiation time 30min, UV fluorescent tube and cultivation surface apart from 3cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the polyoxymethylene cell culturing surfaces of carboxyl grafting.
The polyoxymethylene cell culturing surfaces carboxyl-content that present embodiment prepares is 38.56nmol/cm
2
Embodiment 10
A kind of plasma body-photochemical method at the cell culturing surfaces carboxyl grafting may further comprise the steps:
(1) cell culturing surfaces of polyethylene terephthalate-1,4 cyclohexane dimethanol ester material is put into plasma-reaction-chamber and carry out pre-treatment, the plasma-reaction-chamber working parameter is: power 600W; Pressure 80Pa; Gas flow rate 500cc/min, 35 ℃ of temperature, treatment time 30s;
(2) pretreated cultivation surface is placed on 24h in the air or oxygen;
(3) with monomer vinylformic acid 4.0% (all refer to its mass concentration in solution A, down with), vinylformic acid-2-carboxylic ethyl ester 3.0%, methylacrylic acid 2.0%, N hydroxymethyl acrylamide 2.5%, the N-vinyl pyrrolidone 4.0% formation solution A that is dissolved in the water.Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer, the liquid level layer height is 2mm;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation.UV power of lamp tube 300W, irradiation time 30min, UV fluorescent tube and cultivation surface apart from 3cm;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the polyethylene terephthalate-1,4 cyclohexane dimethanol ester cell culturing surfaces of carboxyl grafting.
The polyethylene terephthalate that present embodiment prepares-1,4 cyclohexane dimethanol ester cell culturing surfaces carboxyl-content is 56.82nmol/cm
2
The foregoing description is a preferred implementation of the present invention; But embodiment of the present invention is not restricted to the described embodiments; Other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; All should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (6)
1. plasma body-photochemical method at the cell culturing surfaces carboxyl grafting is characterized in that may further comprise the steps:
(1) cell culturing surfaces is put into plasma-reaction-chamber and carry out pre-treatment;
(2) pretreated cultivation surface is placed on 25min～24h in the air or oxygen;
(3) with the monomer formation solution A that is dissolved in the water; Solution A is spread into the cultivation surface that step (2) obtains equably form thin liquid level layer;
(4) cultivation surface that is covered with the liquid level layer is carried out the UV irradiation;
(5) cultivation surface is taken out from solution A, water cleans, and obtains the cell culturing surfaces of carboxyl grafting;
The material of said cell culturing surfaces is a kind of in PS, Vilaterm, Vestolen PP 7052, polymethylmethacrylate, polyethylene terephthalate, polycarbonate, polyoxymethylene or polyethylene terephthalate-1,4 cyclohexane dimethanol ester;
The said monomer of step (3) is the two or more mixtures in vinylformic acid, methylacrylic acid, Rocryl 400, maleic anhydride, N-vinyl pyrrolidone, vinylformic acid-2-carboxylic ethyl ester or the N hydroxymethyl acrylamide.
2. plasma body-the photochemical method at the cell culturing surfaces carboxyl grafting according to claim 1; It is characterized in that: the working parameter of the said plasma-reaction-chamber of step (1) is: power 35～600W; Pressure 10～80Pa, gas flow rate 5～500cc/min, 20～35 ℃ of temperature.
3. plasma body-the photochemical method at the cell culturing surfaces carboxyl grafting according to claim 1 is characterized in that: the said pretreated time of step (1) is 30s～10min.
4. plasma body-the photochemical method at the cell culturing surfaces carboxyl grafting according to claim 1 is characterized in that: in the said solution A of step (3), the monomer total mass concentration is 11.5-15.5%.
5. plasma body-the photochemical method at the cell culturing surfaces carboxyl grafting according to claim 1 is characterized in that: the height of the said liquid level layer of step (3) is 0.5～2mm.
6. plasma body-the photochemical method at the cell culturing surfaces carboxyl grafting according to claim 1; It is characterized in that: the described UV irradiation of step (4); Its correlation parameter is: UV power of lamp tube 300～1000W; Irradiation time 2～30min, the distance 3～13cm of UV fluorescent tube and cultivation surface.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110200304A CN102276866B (en) | 2011-07-18 | 2011-07-18 | Plasma-photochemical method for grafting carboxyl on cell culture surface |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110200304A CN102276866B (en) | 2011-07-18 | 2011-07-18 | Plasma-photochemical method for grafting carboxyl on cell culture surface |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102276866A CN102276866A (en) | 2011-12-14 |
| CN102276866B true CN102276866B (en) | 2012-10-03 |
Family
ID=45102630
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110200304A Expired - Fee Related CN102276866B (en) | 2011-07-18 | 2011-07-18 | Plasma-photochemical method for grafting carboxyl on cell culture surface |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102276866B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102533632B (en) * | 2012-01-17 | 2014-10-22 | 海狸纳米科技（苏州）有限公司 | Polypeptide biological nano surface for serum-free cell culture and preparation method |
| CN104946529A (en) * | 2014-03-31 | 2015-09-30 | 海狸纳米科技（苏州）有限公司 | Treatment method for culture bottle capable of preventing liquid drops from hanging at opening |
| CN104974366A (en) * | 2014-04-02 | 2015-10-14 | 苏州科技学院 | Surface treatment method for disposable plastic petri dish |
| CN104788707A (en) * | 2015-04-22 | 2015-07-22 | 广州洁特生物过滤股份有限公司 | Preparation method of super-hydrophilic cell growth surface |
| WO2017051912A1 (en) * | 2015-09-25 | 2017-03-30 | 三菱瓦斯化学株式会社 | Substrate for cell culture, cell culture method using same, cell culture vessel, and use as substrate |
| CN108588006A (en) * | 2018-05-10 | 2018-09-28 | 华东理工大学 | A kind of biological support and its preparation method and application for liver cell dimensional culture |
| TW202334264A (en) * | 2022-02-21 | 2023-09-01 | 穎利科技股份有限公司 | Method for hydrophilic treatment of substrate surface and substrate with hydrophilic surface |
| CN114920972A (en) * | 2022-05-25 | 2022-08-19 | 广州热泉科技有限公司 | Preparation method of temperature-sensitive culture dish |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006058237A2 (en) * | 2004-11-24 | 2006-06-01 | Corning Incorporated | Polymer-coated substrates for binding biomolecules and methods of making and using thereof |
| CN101045797A (en) * | 2007-03-30 | 2007-10-03 | 东华大学 | Preparation of moisture controllable polymer composite film and its application |
| CN101985499A (en) * | 2010-10-28 | 2011-03-16 | 浙江工业大学 | Method for preparing super-hydrophobic surface polypropylene |
| US20110171070A1 (en) * | 2008-05-28 | 2011-07-14 | Forward Electronics Co., Ltd. | Surface-modified sensor device and method for surface-modifying the same |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003045461A1 (en) * | 2001-11-23 | 2003-06-05 | Feg Textiltechnik Forschungs- Und Entwicklungsgesellschaft Mbh | Surface-modified textile product and a corresponding method for surface modification |
-
2011
- 2011-07-18 CN CN201110200304A patent/CN102276866B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006058237A2 (en) * | 2004-11-24 | 2006-06-01 | Corning Incorporated | Polymer-coated substrates for binding biomolecules and methods of making and using thereof |
| CN101045797A (en) * | 2007-03-30 | 2007-10-03 | 东华大学 | Preparation of moisture controllable polymer composite film and its application |
| US20110171070A1 (en) * | 2008-05-28 | 2011-07-14 | Forward Electronics Co., Ltd. | Surface-modified sensor device and method for surface-modifying the same |
| CN101985499A (en) * | 2010-10-28 | 2011-03-16 | 浙江工业大学 | Method for preparing super-hydrophobic surface polypropylene |
Non-Patent Citations (1)
| Title |
|---|
| 王琳琳，黄江，金伟 等.环氧树脂微流控芯片的表面功能化修饰.《高等学校化学学报》.2011,第32卷全文. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102276866A (en) | 2011-12-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102276866B (en) | Plasma-photochemical method for grafting carboxyl on cell culture surface | |
| Lee et al. | Role of reactive gas in atmospheric plasma for cell attachment and proliferation on biocompatible poly ɛ-caprolactone film | |
| Chen et al. | Surface modification of non-woven fabric by DC pulsed plasma treatment and graft polymerization with acrylic acid | |
| JPH02211865A (en) | Support material for culturing cell | |
| NO20083285L (en) | Method and apparatus for precision processing of substrates by means of a laser arranged in a liquid flow, and use thereof | |
| US20140335610A1 (en) | Cell culture substrate, and method for manufacturing same | |
| CN108192034A (en) | The preparation method of high-hydroscopicity gamma-polyglutamic-acid hydrogel material | |
| CN106011834A (en) | Preparation method of strontium chelated sodium alginate coating on titanium surface | |
| CN110355082A (en) | A kind of surface treatment method for reinforcing stainless steel sanitary ware antibacterial durability | |
| CN104788707A (en) | Preparation method of super-hydrophilic cell growth surface | |
| CN104744720A (en) | Preparation method of polyvinylpyrrolidone grafting modified polyethylene terephthalate film | |
| KR101400888B1 (en) | Method of manufacturing surfaces for controlling cell attachment using plasma-treated biopolymer | |
| CN106047692B (en) | A kind of perovskite thin film device and its preparation method and application for visible photic cell harvesting | |
| CN104693277B (en) | A kind of self-assembled short peptide and its application in three-dimensional cell cultivation | |
| CN102533632B (en) | Polypeptide biological nano surface for serum-free cell culture and preparation method | |
| CN106084140A (en) | A kind of polyamide containing guanidino group, its preparation method and the hydrogel prepared by it | |
| CN105908159A (en) | A preparing method of a g-C3N4/FTO composite clear electrically conductive film | |
| JP2008220205A (en) | Container for forming neural stem cell aggregate, method for producing the same and method for preparing neural stem cell aggregate | |
| KR20140128124A (en) | Method for culture of human embryonic stem cells using adhesive heparins | |
| CN106832380A (en) | A kind of method that utilization Laser Processing prepares picture on surface polymer film | |
| JPS63198980A (en) | Base material for cultivating cell | |
| CN109777741B (en) | Method for efficiently utilizing walnut shells | |
| Dahan et al. | Dielectric properties of collagen on plasma modified polyvinylidene fluoride | |
| JP4269256B2 (en) | Nerve cell culture substrate and method for producing the same | |
| JPH05244938A (en) | Cell and its culture |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: BEAVER NANOTECHNOLOGY (SUZHOU) INC. Free format text: FORMER OWNER: BEAVER BIOSCIENCES INC. Effective date: 20131205 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 510663 GUANGZHOU, GUANGDONG PROVINCE TO: 215000 SUZHOU, JIANGSU PROVINCE |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20131205 Address after: Suzhou City, Jiangsu Province, Suzhou Industrial Park 215000 Xinghu Street No. 218 BioBAY A6-101 Patentee after: Beaver Nano-Technologies Co., Ltd. Address before: 510663 Guangdong city of Guangzhou province Guangzhou Science City skim Springs Road No. 3, international business incubator D313 Patentee before: Beaver (Guangzhou) Biological Technology Co., Ltd. |
|
| C56 | Change in the name or address of the patentee | ||
| CP01 | Change in the name or title of a patent holder |
Address after: Suzhou City, Jiangsu Province, Suzhou Industrial Park 215000 Xinghu Street No. 218 BioBAY A6-101 Patentee after: Suzhou beaver Biomedical Engineering Co., Ltd. Address before: Suzhou City, Jiangsu Province, Suzhou Industrial Park 215000 Xinghu Street No. 218 BioBAY A6-101 Patentee before: Beaver Nano-Technologies Co., Ltd. |
|
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20121003 Termination date: 20180718 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |