CN103525700A - Rotational shear force stimulation and electric stimulation combined cell culture device - Google Patents
Rotational shear force stimulation and electric stimulation combined cell culture device Download PDFInfo
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- CN103525700A CN103525700A CN201310414782.9A CN201310414782A CN103525700A CN 103525700 A CN103525700 A CN 103525700A CN 201310414782 A CN201310414782 A CN 201310414782A CN 103525700 A CN103525700 A CN 103525700A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/12—Well or multiwell plates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/48—Holding appliances; Racks; Supports
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
Abstract
The invention discloses a rotational shear force stimulation and electric stimulation combined cell culture device which comprises a cell culture plate, a shaking table, a signal generator and a cell culture box, wherein the cell culture plate is fixed on the shaking table and can provide rotational shear force stimulation to the cells; a needle electrode and an annular electrode are arranged in the cell culture plate; and the signal generator is connected with the needle electrode and the annular electrode through an electrode wire and can provide electric simulation to the cells. According to the invention, a cell culture method under the joint effect of rotational shear force stimulation and electric stimulation is established, and the development of a cell and tissue culture method and related tissue engineering reactors is promoted.
Description
Technical field
The present invention relates to a kind of rotational shear power and electricity combined stimulation cell culture apparatus, belong to cell engineering and tissue engineering technique field.
Background technology
Cell cultures is the importance of Tissue Engineering Study, is keeping under the prerequisite of cell normal physiological function, how economical, stable, obtain a large amount of cells to high-density, becomes one of emphasis of Tissue Engineering Study.Cell is subject to the impact of many factors in vivo, and as stress, micro-electric current, somatomedin etc., these factors have important impact to the growth of cell.Applicable physical stimulation stimulates or electricity irritation can produce promoter action to aspects such as the propagation of cell, differentiation and metabolism as power.
The regulation and control of mechanical environment play key effect in cell, organizational project culturing process, and it is applied to certain shearing force, can keep and further bring into play the function of cell.Research shows after shear action, proliferation of rat osteoblasts ability improves, cytoactive strengthens (Wu Dan etc., 2007), the content of the crosslinking protein on cytoskeleton, α-actin and filamin significantly increases, after hydrodynamic shear effect, they sprawl get Geng Kai (Wesley M et al., 2008) in cell simultaneously; Applicable hydrodynamic shear can promote the expression of scleroblast internal force sensitive gene and synthesizing of associated protein, as immediate-early gene, short inflammation gene expression (Celil A B et al., 2007), osteopontin (Ponik S M et al., 2007) etc., contribute to Matrix Mineralization and bone to rebuild.Rotational shear power also can significantly promote the propagation (Haga M, Yamashita A et al., 2003) of smooth muscle cell.
Except complicated mechanical environment, electricity environment is also one of important microenvironment of cells in vivo growth.By analogue body inner cell, the residing electricity microenvironment of tissue growth, can make cell fast, high-quality amplification.Use electric polypyrrole as culture medium material, neurocyte PC-12 etc. to be cultivated, experiment shows to add electricity irritation and saves than not adding electrical stimulation will nearly double (Schmidt CE, et al., 1997; Ze Zhang, et al., 2007); Pulsed voltage (frequency 20Hz, pulse width 0.4ms) by 1V, 5V, 10V and 15V stimulates scleroblast, finds that cell proliferation situation has had significant lifting (Ercan B, et al., 2008); Electricity irritation can enlarge markedly scleroblast at the unit elongation of nano-material surface, and osteanagenesis and fractures are had to potential using value (Shijun Shao, et al., 2011).
As from the foregoing, loading force stimulates or electricity irritation respectively, can promote the growth of cell.But cell itself is in power, the simultaneous environment of electricity irritation, for maintaining cell normal function, differentiation, secretion etc., is all very important.And existing cell, tissue culture method much all do not have in conjunction with these two kinds of environmental influences.The present invention has considered the impact of the stimulation of rotational shear power and these two kinds of physical stimulation factor cell growth of electricity irritation.
Summary of the invention
The object of the invention is can not provide to cell the problem of rotational shear power, the simultaneous physical environment of electricity irritation in order to solve existing method, cell is placed under the combined action of the stimulation of rotational shear power and electricity irritation, analogue body inner cell living environment, promotes growth and the differentiation of cell.
A kind of rotational shear power of the present invention and electricity combined stimulation cell culture apparatus, comprise Tissue Culture Plate, shaking table, signal generator and cell culture incubator; Tissue Culture Plate is fixed on shaking table, and is connected with signal generator by electrode wires; Shaking table is positioned in cell culture incubator, and signal generator is positioned over outside cell culture incubator; Cell culture incubator both sides are respectively equipped with a duct communicating with the external world, shaking table supply lead in cell culture incubator and electrode wires respectively passing hole channel are connected with power supply and signal generator, cell culture incubator is sealed structure, and cell culture incubator provides the required carbonic acid gas of Growth of Cells, temperature and humidity environment;
In described Tissue Culture Plate, be provided with two metal electrodes, i.e. pin electrode and ring electrode, pin electrode vertically inserts in the culture hole of Tissue Culture Plate by the aperture of Tissue Culture Plate top cover, and ring electrode is placed on Tissue Culture Plate base; Pin electrode is connected with positive pole, the negative pole of signal generator respectively with ring electrode;
When shaking table rotates, can load rotational shear power to the cell in Tissue Culture Plate stimulates; Signal generator can load electricity irritation by electrode pair cell.By changing voltage or the electric current of signal generator, the electricity irritation of different sizes is provided, electricity irritation is constant voltage, pulsed voltage or DC micro-electric stream etc.When working simultaneously, shaking table and signal generator can load rotational shear power and electricity combined stimulation to cell.
The invention has the advantages that:
(1) power electricity combined stimulation loading regime provided by the invention is closer to the internal milieu of Growth of Cells, growth, and the combined action of power electricity can improve Growth of Cells condition, thereby Promote cell's growth, improves cytoactive;
(2) the present invention both can be used for cell, tissue culture, also can be used for cell, tissue growth research, and cell expansion ex vivo is had to positive meaning;
(3) the present invention is simple to operate, and good stability has larger application prospect.
Accompanying drawing explanation
Fig. 1 is the structural representation of a kind of rotational shear power of the present invention and electricity combined stimulation cell culture apparatus;
Fig. 2 is the structural representation of Tissue Culture Plate of the present invention;
In figure:
1-signal generator 2-electrode wires 3-Tissue Culture Plate 4-shaking table 5-cell culture incubator
301-pin electrode 302-ring electrode 303-Tissue Culture Plate top cover 304-Tissue Culture Plate base
Embodiment
Below in conjunction with drawings and Examples, the present invention is described in further detail.
The present invention is a kind of rotational shear power and electricity combined stimulation cell culture apparatus, as shown in Figure 1, comprises signal generator 1, electrode wires 2, Tissue Culture Plate 3, shaking table 4 and cell culture incubator 5;
Tissue Culture Plate 3 is fixed on shaking table 4, is placed in cell culture incubator 5.Electrode wires 2 connects the electrode of Tissue Culture Plate 3 and the positive and negative electrode of signal generator 1.The supply lead of shaking table 4 passes connection power supply from the duct of cell culture incubator 5, and electrode wires 2 passes with signal generator and is connected from the duct of cell culture incubator 5.Cell culture incubator 5 is sealed structure, and the required carbonic acid gas of Growth of Cells, temperature and humidity environment are provided;
The structure of Tissue Culture Plate 3 as shown in Figure 2, inside be provided with two metal electrodes, be pin electrode 301 and ring electrode 302, pin electrode 301 vertically inserts in the culture hole of Tissue Culture Plate 3 by the aperture of Tissue Culture Plate top cover 303, and ring electrode 302 is placed on Tissue Culture Plate base 304; Pin electrode 301 is connected with positive pole, the negative pole of signal generator 1 respectively with ring electrode 302; Cell is seeded in the culture hole of Tissue Culture Plate base 304;
When shaking table 4 rotation, can load rotational shear power to the cell in Tissue Culture Plate 3 stimulates; Signal generator 1 can load electricity irritation by electrode 301 and 302 pairs of cells.When working simultaneously, shaking table 4 and signal generator 1 can load rotational shear power and electricity combined stimulation to cell.
Cell cultures detailed process of the present invention is as follows:
Cell is inoculated in the culture hole of Tissue Culture Plate base 304, ring electrode 302 is placed on Tissue Culture Plate base 304.Cover Tissue Culture Plate top cover 303, from the aperture of top cover 303, insert pin electrode 301.By electrode wires 2, pin electrode 301, ring electrode 302 are connected on the positive and negative electrode of signal generator 1.Tissue Culture Plate 3 is fixed on shaking table 4, by the motion of shaking table 4, provides rotational shear power to stimulate; Signal generator 1 with the interior pin electrode 301 of Tissue Culture Plate 3, ring electrode 302 is connected that electricity irritation is provided, and at loading duration, makes to grow under the environment of cell in rotational shear power and electricity irritation double stimuli, completes the cultivation of cell.
Embodiment:
Under aseptic condition 1-3 days ages SD rat suckling mouse scleroblast by 1 * 10
5/ cm
2density is seeded in the culture hole of Tissue Culture Plate base 304, and preculture, after 3 days, is put into ring electrode 302.Cover Tissue Culture Plate top cover 303, in the aperture of Tissue Culture Plate top cover, insert pin electrode 301.By electrode wires 2, pin electrode 301, ring electrode 302 are connected on the positive and negative electrode of signal generator 1.Tissue Culture Plate 3 is fixed on shaking table 4.Open the shaking table 4 and the signal generator 1 that regulate, make the stimulation of rotational shear power and electricity irritation cooperate with scleroblast, scleroblast is grown under the environment of the stimulation of rotational shear power and electricity irritation double stimuli.
The rotational shear power applying stimulates as 8dyn/cm
2, electricity irritation is electric impulse signal, and amplitude is 150mV, and pulse width is 5ms, and frequency is 1Hz, stimulation time is 12h.
Above-mentioned rotational shear power stimulates, the setting of electrical stimulation parameters is to select according to institute's cultured cells feature difference.
Claims (1)
1. rotational shear power and an electricity combined stimulation cell culture apparatus, is characterized in that: comprise Tissue Culture Plate, shaking table, signal generator and cell culture incubator, Tissue Culture Plate is fixed on shaking table, and be connected with signal generator by electrode wires;
In described Tissue Culture Plate, be provided with two metal electrodes, i.e. pin electrode and ring electrode, pin electrode vertically inserts in the culture hole of Tissue Culture Plate by the aperture of Tissue Culture Plate top cover, and ring electrode is placed on Tissue Culture Plate base; Pin electrode is connected with positive pole, the negative pole of signal generator respectively with ring electrode;
Shaking table is positioned in cell culture incubator, guarantees the growing environment of cell, meets it to temperature, humidity and CO
2requirement; Signal generator is positioned over outside cell culture incubator; When shaking table rotates, can load rotational shear power to the cell in Tissue Culture Plate stimulates; Signal generator can load electricity irritation by electrode pair cell.When working simultaneously, shaking table and signal generator can load rotational shear power and electricity combined stimulation to cell.
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| CN201310414782.9A CN103525700A (en) | 2013-09-12 | 2013-09-12 | Rotational shear force stimulation and electric stimulation combined cell culture device |
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| CN201310414782.9A CN103525700A (en) | 2013-09-12 | 2013-09-12 | Rotational shear force stimulation and electric stimulation combined cell culture device |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109055220A (en) * | 2018-10-12 | 2018-12-21 | 新希望六和股份有限公司 | A kind of circular rotating Cell culture plate structure |
| CN112608845A (en) * | 2021-01-11 | 2021-04-06 | 兰州大学第一医院 | Bionic stimulation device for mechanical pressure of stem cells |
| US20220064583A1 (en) * | 2020-09-02 | 2022-03-03 | Samsung Electronics Co., Ltd. | Electrical stimulation device |
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| CN101892125A (en) * | 2010-07-16 | 2010-11-24 | 黄山富溪源茶油开发有限公司 | Efficient winterization device and winterization method of tea oil |
| CN102559492A (en) * | 2011-12-15 | 2012-07-11 | 北京航空航天大学 | Cell culture device capable of loading shear force stimulation and electrical stimulation jointly in parallel direction |
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2013
- 2013-09-12 CN CN201310414782.9A patent/CN103525700A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101892125A (en) * | 2010-07-16 | 2010-11-24 | 黄山富溪源茶油开发有限公司 | Efficient winterization device and winterization method of tea oil |
| CN102559492A (en) * | 2011-12-15 | 2012-07-11 | 北京航空航天大学 | Cell culture device capable of loading shear force stimulation and electrical stimulation jointly in parallel direction |
Non-Patent Citations (2)
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| 王华: "剪切力对体外培养人软骨细胞代谢的影响", 《中国博士学位论文全文数据库》 * |
| 贾璐等: "力/电联合刺激细胞培养体系搭建及装置研制", 《2009年全国大学生创新性实验计划项目成果展示》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109055220A (en) * | 2018-10-12 | 2018-12-21 | 新希望六和股份有限公司 | A kind of circular rotating Cell culture plate structure |
| CN109055220B (en) * | 2018-10-12 | 2021-09-17 | 新希望六和股份有限公司 | Circular rotatory cell culture plate structure |
| US20220064583A1 (en) * | 2020-09-02 | 2022-03-03 | Samsung Electronics Co., Ltd. | Electrical stimulation device |
| US11952565B2 (en) * | 2020-09-02 | 2024-04-09 | Samsung Electronics Co., Ltd. | Electrical stimulation device |
| CN112608845A (en) * | 2021-01-11 | 2021-04-06 | 兰州大学第一医院 | Bionic stimulation device for mechanical pressure of stem cells |
| CN112608845B (en) * | 2021-01-11 | 2023-07-25 | 兰州大学第一医院 | Stem cell mechanical pressure bionic stimulation device |
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