CN105505773A - Magnetic-electric stimulation cell culture device - Google Patents

Magnetic-electric stimulation cell culture device Download PDF

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Publication number
CN105505773A
CN105505773A CN201510999565.XA CN201510999565A CN105505773A CN 105505773 A CN105505773 A CN 105505773A CN 201510999565 A CN201510999565 A CN 201510999565A CN 105505773 A CN105505773 A CN 105505773A
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magnetic
electric
cell
tissue culture
cell culture
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CN201510999565.XA
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李萍
张书培
李昆
牛旭锋
樊瑜波
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Beihang University
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Beihang University
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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
    • 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
    • C12M35/06—Magnetic means

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  • General Engineering & Computer Science (AREA)
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  • Cell Biology (AREA)
  • Physics & Mathematics (AREA)
  • Electromagnetism (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
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Abstract

The invention provides a magnetic-electric stimulation cell culture device capable of being used for loading magnetic-electric stimulation on cells. The device comprises a cell culture board, a magnetic-electric double-function fiber membrane, a pulse electric signal emission source, platinum sheet electrodes and magnets. The magnetic-electric double-function fiber membrane is arranged in a cell culture board base. A pair of parallel platinum sheet electrodes is inserted into the portion, corresponding to each cell culture hole, of a cell culture board cover, the lower ends of the platinum sheet electrodes abut against the bottom of the pore board and make contact with the magnetic-electric double-function fiber membrane, the upper ends of the platinum sheet electrodes are connected with the pulse electric signal emission source through wires, and electric stimulation can be loaded on the cells by switching on the pulse electric signal emission source. A pair of ferrite magnets is placed on the left side and the right side of the cell culture board base in parallel, and magnetic stimulation can be loaded on the cells. The magnetic-electric stimulation cell culture device can load electric stimulation and magnetic stimulation on the cells at the same time, and development of a cell and tissue culture method can be promoted.

Description

A kind of magnetic-Electrical stimulation cell culture device
Technical field
The present invention relates to a kind of magnetic-Electrical stimulation cell culture device based on support, belong to field of cell culture, the research of particularly magnetic-electricity irritation cell growth impact.
Background technology
The growing environment of cell is a very complicated microenvironment; except being subject to the chemical molecular signal, biotic factor signal, mechanical stimulation etc. in environment; cell usually can be subject to the stimulation in micro-electric current and magnetic field in biological tissue; such as bone and its cells is subject to piezoelectric current because organism motion produces, electricity irritation and Neural stem cell; not only have great significance to the growth of cell, differentiation, be also the prerequisite of its function of cells play simultaneously.
There are some researches prove, the normal conduction that a lot of cells in human body all rely on electric charge plays its physiological function, and such as the disappearance of a large amount of normal function type myocardial cell can cause the improper electrical conductive behavior of myocardial cell, finally often causes myocardial infarction.Electricity irritation is of great significance the same tool of histocyte that scleroblast, neurocyte etc. are subject to microelectric current impact in body.And magnetic field can effectively prevent bone density from reducing, promote bone injury healing.
The people such as Chun develop a kind of conductive fabric, and the myocardial cell on conducting film is interconnected to form cell mass under adding electricity irritation outside, and group's inner cell is along material orderly direction growth.After cultivating, can clearly observe cell articulin 43 in cell mass, all myocardial cells can synchronously beat.
The mode that the people such as Zhuang use condenser coupling to stimulate, mouse bone-forming cell is loaded with to the electricity irritation of 20mV/cm, result shows, electricity irritation facilitates osteoblastic differentiation, and scleroblast DNA resultant quantity increases to some extent.
The people such as Shan are by the fixing Fe of blended Poly-L-lactic acid (PLLA) 3o 4nanoparticle has prepared a kind of base material-magnetic Fe for osteanagenesis 3o 4/ PLLA composite nano fiber, and the cytotoxicity of research material.The Fe of different content in composite electrostatic spinning nanofiber 3o 4nanoparticle is dispersed along fiber direction.Obtained Fe 3o 4/ PLLA conjugated fibre shows with the positively related superparamagnetism of magnetic particle content.Fe 3o 4/ PLLA matrix material does not have purer PLLA to show larger cytotoxicity, on the contrary, but shows cell adhesion and the better promoter action of propagation.
But what at present investigator paid close attention to is single physical stimulation mostly, be difficult to Reality simulation biological tissue environment in all directions and investigate that cell is bred, broken up under multiple stimulation, impact that pattern and migratory behaviour are subject to.Therefore research can be that the device tool that cell loads magnetic-electricity combined stimulation is simultaneously of great significance.
Summary of the invention
The object of the invention is to overcome existing apparatus and can not investigate cell affected deficiency under multiple stimulation, providing can be the device that cell loads magnetic, electricity combined stimulation simultaneously.
A kind of magnetic-Electrical stimulation cell culture device, contrive equipment comprises Tissue Culture Plate, magnetic-electric difunctional tunica fibrosa, pulse electrical signal emissive source, platinum plate electrode and magnet; Magnetic-electric difunctional tunica fibrosa is placed in Tissue Culture Plate base; Tissue Culture Plate lid each cell culture well corresponding vertically inserts pair of parallel platinum plate electrode, and platinum plate electrode upper end connects pulse electrical signal emissive source by wire, and platinum plate electrode lower end near Tissue Culture Plate base, and directly contacts magnetic-electric difunctional tunica fibrosa.Described magnetic-electric difunctional tunica fibrosa is poly (glycolide-lactide) (PLGA)/Fe 3o 4the outer in-situ polymerization of magnetic Nano fiber membrane gathers 3,4-rthylene dioxythiophene conducting polymer coating (PEDOT/Fe 3o 4/ PLGA tunica fibrosa), there is good magnetic and conductivity.Be placed on the Tissue Culture Plate base left and right sides by parallel for a pair ferrite magnet (0.1-1.0T), can be cell and Neural stem cell is provided.
When starting impulse electrical signal emissive source, electricity irritation can be loaded to the cell on magnetic-electric difunctional tunica fibrosa; When not starting impulse electrical signal emissive source, and when a pair ferrite magnet is placed in the Tissue Culture Plate base left and right sides, Neural stem cell can be loaded to the cell on magnetic-electric difunctional tunica fibrosa; When starting impulse electrical signal emissive source, and when placing a pair ferrite magnet, Neural stem cell and electricity irritation can be loaded to the cell on magnetic-electric difunctional tunica fibrosa simultaneously.
The invention provides a kind of magnetic-Electrical stimulation cell culture device, can be used for the impact studying electricity irritation, Neural stem cell and magnetic-electricity combined stimulation on cell proliferation, differentiation, pattern.
The invention has the advantages that:
(1) device of the present invention carries simple, and cost is low, is easy to cleaning and sterilizing;
(2) electric signal parameter used in the present invention and magnitude of field intensity adjustable;
(3) the present invention can load electricity irritation and Neural stem cell to cell simultaneously, simulates complex environment in organism better, significant on the impact of the cell proliferation of research different stimulated, differentiation, pattern.
Accompanying drawing explanation
Fig. 1 is magnetic of the present invention-Electrical stimulation cell culture device structural representation, in figure:
1-pulse electrical signal emissive source 2-Tissue Culture Plate lid 3-platinum plate electrode 4-magnet 5-Tissue Culture Plate base
Embodiment
Below in conjunction with drawings and Examples, the present invention is described in further detail.
The present invention is magnetic-Electrical stimulation cell culture device, as shown in Figure 1, pulse electrical signal emissive source (1), Tissue Culture Plate lid (2), platinum plate electrode (3), magnet (4) and Tissue Culture Plate base (5) is comprised;
Magnetic-Electrical stimulation cell culture device uses medical alcohol wiping, and soak 30 minutes, ultra violet lamp 2h carries out aseptically process;
Magnetic-electric difunctional tunica fibrosa is cut into circle, puts into Tissue Culture Plate base (5);
Cell good for growth conditions is seeded on the magnetic-electric difunctional tunica fibrosa in Tissue Culture Plate base (5);
Tissue Culture Plate lid (2) is vertically inserted in platinum plate electrode (3) parallel placement, the magnetic in lower end direct exposing cell culture plate base (5)-electric difunctional tunica fibrosa; Upper end connects pulse electrical signal emissive source (1) by wire and arranges suitable electric signal parameter;
Magnetic-Electrical stimulation cell culture device is placed in cell culture incubator, open the pulse electrical signal emissive source (1) be connected with platinum plate electrode (3), a pair rectangular parallelepiped ferrite magnet (4) is parallel is placed on Tissue Culture Plate base (5) left and right sides, can load magnetic-electricity irritation to cell.
Embodiment 1
Device is as shown in Figure 1 adopted to load magnetic-electricity irritation to scleroblast MC3T3-E1.
By PEDOT/Fe 3o 4it is that the circle of 1cm is placed in 24 porocyte culture plate bases that/PLGA magnetic-electric difunctional tunica fibrosa is cut into diameter.Scleroblast MC3T3-E1 is with 1 × 10 5the density in individual/hole is seeded in PEDOT/Fe 3o 4on/PLGA magnetic-electric difunctional tunica fibrosa.Tissue Culture Plate is steadily placed on CO 2cultivate 30min in cell culture incubator, start, after attaching, slowly to add the MEM perfect medium of 1mL in each orifice plate, be placed in CO until cell 2cultivate in cell culture incubator, example of spatial compartmentalizationis.After cell inoculates 24 hours, open pulse electrical signal emissive source, pulse electrical signal emissive source parameter is amplitude is 500mV, and frequency is 20Hz, and dutycycle is 0.1%, and waveform is rectangular pulse signal.A pair ferrite magnet (15 × 88 × 40cm, 0.72T) is parallel is placed on the Tissue Culture Plate base left and right sides, loads Neural stem cell to cell.When pulse electrical signal emissive source starts and places ferrite magnet, the scleroblast MC3T3-E1 in device can be subject to magnetic-electricity combined stimulation, and detects in sampling in the 1st, 3,5 day.
Embodiment 2
Device is as shown in Figure 1 adopted to load magnetic-electricity irritation to Human umbilical vein endothelial cells HUVEC.
By PEDOT/Fe 3o 4it is that the circle of 1cm is placed in 24 porocyte culture plate bases that/PLGA magnetic-electric difunctional tunica fibrosa is cut into diameter.HUVEC is with 1 × 10 5the density in individual/hole is seeded on magnetic-electric difunctional tunica fibrosa.Tissue Culture Plate is steadily placed on CO 2cultivate 30min in cell culture incubator, start, after attaching, slowly to add the DMEM perfect medium of 1mL in each orifice plate, be placed in CO until cell 2cultivate in cell culture incubator, example of spatial compartmentalizationis.After cell inoculates 24 hours, open pulse electrical signal emissive source, pulse electrical signal emissive source parameter is amplitude is 200mV, and frequency is 20Hz, and dutycycle is 0.1%, and waveform is rectangular pulse signal.A pair ferrite magnet (15 × 88 × 40cm, 0.72T) is parallel is placed on the Tissue Culture Plate base left and right sides, loads Neural stem cell to cell.When pulse electrical signal emissive source starts and places ferrite magnet, the HUVEC in device can be subject to magnetic-electricity combined stimulation, and detects in sampling in the 1st, 3,5 day.

Claims (5)

1. magnetic-Electrical stimulation cell culture device, is characterized in that: comprise Tissue Culture Plate, magnetic-electric difunctional tunica fibrosa, pulse electrical signal emissive source, platinum plate electrode and magnet; Magnetic-electric difunctional tunica fibrosa is placed in Tissue Culture Plate base; Platinum plate electrode inserts Tissue Culture Plate lid each cell culture well place corresponding; Platinum plate electrode is connected with pulse electrical signal emissive source; Magnet is positioned at the Tissue Culture Plate base left and right sides.
2. device according to claim 1, it is characterized in that, platinum plate electrode covers corresponding each cell culture well from Tissue Culture Plate and vertically inserts, contact with the magnetic in Tissue Culture Plate base-electric difunctional tunica fibrosa, platinum plate electrode upper end connects pulse electrical signal emissive source by wire, often pair of opposing parallel placement of platinum plate electrode.
3. device according to claim 1, it is characterized in that, described magnetic-electric difunctional tunica fibrosa is Fe 3o 4the outer in-situ polymerization of/poly (glycolide-lactide) magnetic Nano fiber membrane gathers 3,4-rthylene dioxythiophene conducting polymer coating, has magnetic and conductivity.
4. device according to claim 1, it is characterized in that, described magnet is a pair ferrite magnet, is parallelly placed on the Tissue Culture Plate base left and right sides, provides static magnetic field (0.1 ~ 1.0T).
5. device according to claim 1, it is characterized in that, described magnetic-electricity irritation is for be seeded on magnetic-electric difunctional tunica fibrosa by cell, and magnet is placed on the Tissue Culture Plate base left and right sides, open the pulse electrical signal emissive source be connected with platinum plate electrode, magnetic-electricity irritation is loaded to cell.
CN201510999565.XA 2015-12-28 2015-12-28 Magnetic-electric stimulation cell culture device Pending CN105505773A (en)

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CN110694115A (en) * 2019-10-22 2020-01-17 上海交通大学医学院附属第九人民医院 Method for constructing tendon tissue in vitro, and biological material and application thereof
ES2793098A1 (en) * 2020-10-13 2020-11-12 Univ Murcia Device for the in vitro application of electrolysis using needles (Machine-translation by Google Translate, not legally binding)
WO2020258083A1 (en) * 2019-06-26 2020-12-30 巽晨国际股份有限公司 Cell culture method

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020258083A1 (en) * 2019-06-26 2020-12-30 巽晨国际股份有限公司 Cell culture method
CN110694115A (en) * 2019-10-22 2020-01-17 上海交通大学医学院附属第九人民医院 Method for constructing tendon tissue in vitro, and biological material and application thereof
CN110694115B (en) * 2019-10-22 2022-03-01 上海交通大学医学院附属第九人民医院 Method for constructing tendon tissue in vitro, and biological material and application thereof
ES2793098A1 (en) * 2020-10-13 2020-11-12 Univ Murcia Device for the in vitro application of electrolysis using needles (Machine-translation by Google Translate, not legally binding)
WO2022079330A1 (en) * 2020-10-13 2022-04-21 Universidad De Murcia Device for the in vitro application of needle electrolysis

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Application publication date: 20160420