CN105695329B - High-flux electric stimulates the experimental provision and method of Cell differentiation inducing activity and medicine controlled releasing - Google Patents
High-flux electric stimulates the experimental provision and method of Cell differentiation inducing activity and medicine controlled releasing Download PDFInfo
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- CN105695329B CN105695329B CN201610214618.7A CN201610214618A CN105695329B CN 105695329 B CN105695329 B CN 105695329B CN 201610214618 A CN201610214618 A CN 201610214618A CN 105695329 B CN105695329 B CN 105695329B
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- 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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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
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Abstract
A kind of high-flux electric stimulates the experimental provision of Cell differentiation inducing activity and medicine controlled releasing, and device includes electrical stimulation signal generator and cell culture apparatus.The composition of electrical stimulation signal generator is：Single-chip minimum system, digital analog converter, Signal-regulated kinase single-pole double-throw switch (SPDT) moved end be sequentially connected；There are upper cover plate, one end of upper cover plate connection electric wire to be connected with the titanium alloy electrode that plate is hung in cell culture apparatus at the top of the cell culture apparatus, the other end is connected with the first non-moving end of switch；The bottom plate of the cell culture apparatus is drug release plate, and drug release plate is connected with the ground terminal of the second non-moving end of switch and Signal-regulated kinase simultaneously；It is equipped on drug release plate and carries medicine conductive polymer film.It can generate optional frequency and the electrical stimulation signal of amplitude, moreover it is possible to while drug is added, to carry out the screening of the high-throughput electrical stimulation signal and drug during stem cell differentiation in vitro, more extensive, reliable experimental basis is provided for stimulation therapy.
Description
Technical field
The present invention relates to a kind of electro photoluminescence Cell differentiation inducing activity and the experimental provisions and method of medicine controlled releasing.
Background technology
In organizational project, the external functionalization culture of cell, tissue has become the core of organizational project, and provide with
Under the premise of the internal similar living environment of cell, how by controlling environmental stimuli or medicine controlled releasing come inducing cell towards setting side
It is most important for the dimensional culture and functionalization of cell, tissue to differentiation.In recent years, electronic stimulation development is very rapid,
From single electrical stimulation solve the problems, such as muscular movement develop to adjust stem cell differentiation, start limbs and spinal cord regeneration, lure
Lead the generation of dystopy eyeball and control tumour growth etc..
By external electro photoluminescence cell culture test device, can test out electrical stimulation signal and cell the speed of growth,
Power of regeneration, the effect broken up to specific direction and relationship improve electro photoluminescence and control to provide experimental basis for electronic stimulation
The effect for the treatment of.But existing external electro photoluminescence cell culture test device, the electrical stimulation signal provided is typically voltage constant
Drug can not be also added in direct current in culture environment, thus can not be to the ac signal and cell of various frequencies and amplitude
The speed of growth, power of regeneration, to specific direction break up effect and relationship studied and tested；It can not be to exchanging telecommunications
Number with drug to the speed of growth of cell, power of regeneration, to specific direction break up cooperate with, inhibition and relationship are tested
And research.Hinder the further development of electronic stimulation.
Invention content
The first object of the present invention is to provide the experiment dress of a kind of high-flux electric stimulation Cell differentiation inducing activity and medicine controlled releasing
It sets, which can both generate the electrical stimulation signal of wide range of frequencies and amplitude, moreover it is possible to while drug is added, to be done in vitro
The screening of high-throughput electrical stimulation signal and drug in cell differentiation procedure provides more extensive, reliable real for stimulation therapy
Test foundation.
Technical solution is used by the present invention realizes above-mentioned first purpose：A kind of high-flux electric stimulation Cell differentiation inducing activity
And the experimental provision of medicine controlled releasing, including electrical stimulation signal generator and cell culture apparatus, it is characterised in that：
The composition of the electrical stimulation signal generator is：The general end of single-chip minimum system and digital analog converter it is defeated
Enter end to be connected, the output end of digital analog converter is connected with the input terminal of Signal-regulated kinase；The output end of Signal-regulated kinase with
The moved end of single-pole double-throw switch (SPDT) K is connected；
Upper cover plate is stamped at the top of the cell culture apparatus, upper cover plate connects electric wire, upper cover plate connection embedded with upper cover plate
One end of electric wire is pierced by the bottom of upper cover plate and is connected with the titanium alloy electrode of plate, and titanium alloy electrode connects electricity by upper cover plate
Line is hung in the inner cavity of cell culture apparatus；The other end of the upper cover plate connection electric wire is pierced by side and the hilted broadsword of upper cover plate
The first non-moving end of commutator K is connected；
The bottom plate of the cell culture apparatus is drug release plate, and the drug release plate is titanium alloy sheet；Drug is released
Put plate and be welded with drug release switching plate connecting line and drug release plate ground wire, the other end of drug release switching plate connecting line with
The second non-moving end of single-pole double-throw switch (SPDT) K is connected；The other end of drug release plate ground wire is connected with the ground terminal of Signal-regulated kinase；
The upper surface of the drug release plate, which is placed with, carries medicine conductive polymer film.
The second object of the present invention is to provide a kind of using above-mentioned high-flux electric stimulation Cell differentiation inducing activity and drug control
The experimental provision released carries out the experimental method of high-flux electric stimulation Cell differentiation inducing activity and medicine controlled releasing, and this method can be in experiment
Generate the electrical stimulation signal of optional frequency and amplitude, moreover it is possible to while drug be added, during carrying out stem cell differentiation in vitro
High-throughput electrical stimulation signal and drug screening, to provide more extensive, reliable experimental basis for stimulation therapy.
The present invention realizes above-mentioned second purpose the technical scheme adopted is that a kind of lured using above-mentioned high-flux electric stimulation
Guided cell, which breaks up, and the experimental provision of medicine controlled releasing carries out high-flux electric stimulates the experiment side of Cell differentiation inducing activity and medicine controlled releasing
Method, operation are：
A, cell inoculation：Stem cell is seeded on the load medicine conductive polymer film on drug release plate, and to cell
Culture solution is added in incubator, and culture solution is made to flood titanium alloy electrode；
B, signal occurs：Electrical stimulation signal generator is opened, electrical stimulation signal generator is made to generate setpoint frequency and amplitude
Square wave, sine wave or sawtooth wave electrical stimulation signal, or the direct current that makes electrical stimulation signal generator generate setting voltage value
Electrical stimulation signal；
C, electro photoluminescence medicine controlled releasing：Operation single-pole double-throw switch (SPDT) makes its moved end be connected to the first non-moving end, electrical stimulation signal
The electrical stimulation signal that generator generates passes sequentially through titanium alloy electrode, culture solution, carries medicine conductive polymer film and drug release
Plate returns to electrical stimulation signal generator；The drug in medicine conductive polymer film is carried, is released under the stimulation control of electrical stimulation signal
It is put into culture solution, to carry out electro photoluminescence medicine controlled releasing；
D, electro photoluminescence cell differentiation：After the electro photoluminescence of C steps was carried out to the time of setting, operation single-pole double-throw switch (SPDT) makes it
Moved end is connected to the second non-moving end, electrical stimulation signal generator generate electrical stimulation signal by carry medicine conductive polymer film and
Drug release plate returns to electrical stimulation signal generator；The stem cell carried on medicine conductive polymer film is inoculated in by electro photoluminescence to be believed
Number stimulation, induction generate cell differentiation.
E, terminate：After the electro photoluminescence cell differentiation of D steps reaches the time of setting, electrical stimulation signal generator is closed, it will
Stem cell in cell culture apparatus is taken out, and experiment terminates.
The working process and principle of electrical stimulation signal generator of the present invention are：
It is generated and setpoint frequency and the square wave of amplitude, sine wave, sawtooth wave or setting voltage value by single-chip minimum system
The corresponding digital signal of direct current, which is converted to analog signal by digital analog converter, then through signal condition mould
Block conditioning be the square wave of setpoint frequency and amplitude, sine wave, sawtooth wave or set voltage value direct current electrical stimulation signal, most
Cell culture apparatus is conveyed to by single-pole double-throw switch (SPDT) K afterwards.
Compared with prior art, the beneficial effects of the present invention are：
The device of the invention and method can accurately control generate variety classes, amplitude and frequency electrical stimulation signal, can
Under the conditions of mutually synthermal, humidity and media environment, research variety classes, amplitude and frequency voltage/current stimulus signal
With cell differentiation and the relationship of medicine controlled releasing, to carry out the high-throughput electrical stimulation signal during stem cell differentiation in vitro with
The screening of drug provides more extensive, reliable experimental basis for stimulation therapy.
Further, cell culture apparatus of the invention is 1~200.
The present invention can be with the subsynchronous high-throughput Cell differentiation inducing activity and medicine controlled releasing for carrying out a variety of drugs and stem cell
Experiment more efficiently can quickly filter out drug and electrical stimulation signal most beneficial for cell differentiation.
The present invention is described in further details with reference to the accompanying drawings and detailed description：
Description of the drawings
Fig. 1 is the overall structure diagram of apparatus of the present invention.
The circuit diagram of Fig. 2 electrical stimulation signal generators.
Specific implementation mode
Fig. 1-2 shows, a kind of specific implementation mode of the invention is, a kind of 1. high-flux electric stimulation Cell differentiation inducing activities and
The experimental provision of medicine controlled releasing, including electrical stimulation signal generator 1 and cell culture apparatus 7, it is characterised in that：
The composition of the electrical stimulation signal generator 1 is：The general end of single-chip minimum system MCU and digital analog converter
(DAC) input terminal is connected, and the output end of digital analog converter (DAC) is connected with the input terminal of Signal-regulated kinase SP；Signal tune
The output end of reason module SP is connected with the moved end Kc of single-pole double-throw switch (SPDT) K；
Upper cover plate 6 is stamped at the top of the cell culture apparatus 7, and upper cover plate 6 is embedded with upper cover plate connection electric wire (4), upper cover
One end of plate connection electric wire (4) is pierced by the bottom of upper cover plate 6 and is connected with the titanium alloy electrode 8 of plate, and titanium alloy electrode 8 passes through
Upper cover plate connection electric wire (4) is hung in the inner cavity of cell culture apparatus 7；The other end of the upper cover plate connection electric wire (4) is worn
The side for going out upper cover plate 6 is connected with the first non-moving end (Ka) of single-pole double-throw switch (SPDT) K；
The bottom plate of the cell culture apparatus 7 is drug release plate 9, and the drug release plate 9 is titanium alloy sheet；Drug
Release board 9 is welded with drug release switching plate connecting line 5 and drug release plate ground wire 3, drug release switching plate connecting line 5 it is another
One end is connected with the second non-moving end Kb of single-pole double-throw switch (SPDT) K；The other end of drug release plate ground wire 3 and Signal-regulated kinase SP
Ground terminal be connected；The upper surface of the drug release plate 9, which is placed with, carries medicine conductive polymer film 11.
The cell culture apparatus 7 of this example is 1~200.
It is a kind of to make the high-flux electric of this example that the experimental provision of Cell differentiation inducing activity and medicine controlled releasing be stimulated to carry out high-flux electric
Stimulating the experimental method of Cell differentiation inducing activity and medicine controlled releasing, operation is：
A, cell inoculation：Stem cell is seeded on the load medicine conductive polymer film 11 on drug release plate 9, and to thin
Culture solution is added in born of the same parents' incubator 7, and culture solution is made to flood titanium alloy electrode 8；
B, signal occurs：Open electrical stimulation signal generator 1, make electrical stimulation signal generator (1) generate setpoint frequency and
The square wave of amplitude, the electrical stimulation signal of sine wave or sawtooth wave, or electrical stimulation signal generator is made to generate the straight of setting voltage value
The electrical stimulation signal of stream；
C, electro photoluminescence medicine controlled releasing：Operation single-pole double-throw switch (SPDT) makes its moved end Kc be connected to the first non-moving end (Ka), electricity thorn
The electrical stimulation signal that energizing signal generator 1 generates passes sequentially through titanium alloy electrode 8, culture solution, carries medicine conductive polymer film 11
And drug release plate 9 returns to electrical stimulation signal generator 1；The drug in medicine conductive polymer film 11 is carried, in electrical stimulation signal
Stimulation control under be discharged into culture solution, to carry out electro photoluminescence medicine controlled releasing；
D, electro photoluminescence cell differentiation：After the electro photoluminescence of C steps was carried out to the time of setting, operation single-pole double-throw switch (SPDT) makes it
Moved end Kc is connected to the second non-moving end (Kb), and the electrical stimulation signal that electrical stimulation signal generator 1 generates is by carrying medicine conductive polymer
Sub- film 11 and drug release plate 9 return to electrical stimulation signal generator 1；It is inoculated in dry thin on load medicine conductive polymer film 11
Born of the same parents are stimulated by electrical stimulation signal, induce generation cell differentiation.
E, terminate：After the electro photoluminescence cell differentiation of D steps reaches the time of setting, electrical stimulation signal generator 1 is closed, it will
Stem cell in cell culture apparatus 7 is taken out, and experiment terminates.
Claims (3)
1. the experimental provision of a kind of high-flux electric stimulation Cell differentiation inducing activity and medicine controlled releasing, including electrical stimulation signal generator
(1) and cell culture apparatus (7), it is characterised in that：
The composition of the electrical stimulation signal generator (1) is：The general end of single-chip minimum system (MCU) and digital analog converter
(DAC) input terminal is connected, and the output end of digital analog converter (DAC) is connected with the input terminal of Signal-regulated kinase (SP)；Signal
The output end of conditioning module (SP) is connected with the moved end (Kc) of single-pole double-throw switch (SPDT) (K)；
Upper cover plate (6) is stamped at the top of the cell culture apparatus (7), upper cover plate (6) connects electric wire (4) embedded with upper cover plate, on
One end of cover board connection electric wire (4) is pierced by the bottom of upper cover plate (6) and is connected with the titanium alloy electrode (8) of plate, and titanium alloy electricity
Pole (8) connects electric wire (4) by upper cover plate and hangs in the inner cavity of cell culture apparatus (7)；The upper cover plate connection electric wire (4)
The other end be pierced by the side of upper cover plate (6) and be connected with the first non-moving end (Ka) of single-pole double-throw switch (SPDT) (K)；
The bottom plate of the cell culture apparatus (7) is drug release plate (9), and the drug release plate (9) is titanium alloy sheet；Medicine
Object release board (9) is welded with drug release switching plate connecting line (5) and drug release plate ground wire (3), and drug release switching plate connects
The other end of wiring (5) is connected with the second non-moving end (Kb) of single-pole double-throw switch (SPDT) (K)；Drug release plate ground wire (3) it is another
End is connected with the ground terminal of Signal-regulated kinase (SP)；The upper surface of the drug release plate (9), which is placed with, carries medicine conductive polymer
Sub- film (11).
2. the experimental provision of high-flux electric stimulation Cell differentiation inducing activity according to claim 1 and medicine controlled releasing, feature
It is：The cell culture apparatus (7) is 1~200.
3. a kind of experimental provision using high-flux electric stimulation Cell differentiation inducing activity and medicine controlled releasing described in claim 1 carries out
High-flux electric stimulates the experimental method of Cell differentiation inducing activity and medicine controlled releasing, and operating is：
A, cell inoculation：Stem cell is seeded on the load medicine conductive polymer film (11) on drug release plate (9), and to thin
Culture solution is added in born of the same parents' incubator (7), and culture solution is made to flood titanium alloy electrode (8)；
B, signal occurs：Electrical stimulation signal generator (1) is opened, electrical stimulation signal generator (1) is made to generate setpoint frequency and width
The square wave of value, the electrical stimulation signal of sine wave or sawtooth wave, or electrical stimulation signal generator is made to generate the direct current for setting voltage value
Electrical stimulation signal；
C, electro photoluminescence medicine controlled releasing：Operation single-pole double-throw switch (SPDT) (K) makes its moved end (Kc) be connected to the first non-moving end (Ka), electricity thorn
It is thin that the electrical stimulation signal that energizing signal generator (1) generates passes sequentially through titanium alloy electrode (8), culture solution, load medicine conducting polymer
Film (11) and drug release plate (9) return to electrical stimulation signal generator (1)；The drug in medicine conductive polymer film (11) is carried,
It is discharged into culture solution under the stimulation control of electrical stimulation signal, to carry out electro photoluminescence medicine controlled releasing；
D, electro photoluminescence cell differentiation：After the electro photoluminescence of C steps was carried out to the time of setting, operation single-pole double-throw switch (SPDT) makes its moved end
(Kc) it is connected to the second non-moving end (Kb), the electrical stimulation signal that electrical stimulation signal generator (1) generates is by carrying medicine conductive polymer
Sub- film (11) and drug release plate (9) return to electrical stimulation signal generator (1)；It is inoculated in and carries medicine conductive polymer film (11)
On stem cell stimulated by electrical stimulation signal, induce generate cell differentiation；
E, terminate：After the electro photoluminescence cell differentiation of D steps reaches the time of setting, electrical stimulation signal generator (1) is closed, it will be thin
Stem cell in born of the same parents' incubator (7) is taken out, and experiment terminates.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610214618.7A CN105695329B (en) | 2016-04-08 | 2016-04-08 | High-flux electric stimulates the experimental provision and method of Cell differentiation inducing activity and medicine controlled releasing |
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| CN201610214618.7A CN105695329B (en) | 2016-04-08 | 2016-04-08 | High-flux electric stimulates the experimental provision and method of Cell differentiation inducing activity and medicine controlled releasing |
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| CN105695329A CN105695329A (en) | 2016-06-22 |
| CN105695329B true CN105695329B (en) | 2018-08-17 |
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| CN114621947B (en) * | 2022-03-30 | 2023-09-08 | 中国科学院深圳先进技术研究院 | Stem cell nerve differentiation regulation and control technology based on electrical stimulation |
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