CN105695329A - Experimental device and method for high-throughput electrical-stimulation cell differentiation induction and drug controlled release - Google Patents
Experimental device and method for high-throughput electrical-stimulation cell differentiation induction and drug controlled release Download PDFInfo
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- CN105695329A CN105695329A CN201610214618.7A CN201610214618A CN105695329A CN 105695329 A CN105695329 A CN 105695329A CN 201610214618 A CN201610214618 A CN 201610214618A CN 105695329 A CN105695329 A CN 105695329A
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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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
The invention provides an experimental device for high-throughput electrical-stimulation cell differentiation induction and drug controlled release. The device comprises an electrical stimulation signal generator and a cell culture apparatus, wherein the electrical stimulation signal generator comprises a single-chip microcomputer minimum system, a digital-analog converter and a signal regulating module; the output end of the signal regulating module is connected with the running end of a single-pole double-throw switch; an upper cover plate is arranged at the top of the cell culture apparatus; one end, connected with an electric wire, of the upper cover plate is connected with a titanium alloy electrode which is suspended in the cell culture apparatus, while the other end of the upper cover plate is connected with a first static end of a switch; the bottom plate of the cell culture apparatus is a drug release plate which is simultaneously connected with the second static end of the switch and the ground end of the signal regulating module; and a drug-loading conductive high-molecular film is arranged on the drug release plate. The experimental device can be used for generating electrical stimulation signals with random frequencies and amplitudes, and can be added with drugs to carry out high-throughput electrical stimulation signal and drug screening in the process of peripheral stem cell differentiation, so that a wide and reliable experiment basis is provided to stimulation therapy.
Description
Technical field
The present invention relates to the experimental provision of a kind of electricity irritation Cell differentiation inducing activity and medicine controlled releasing and method。
Background technology
In organizational project, cell, tissue external functionalization cultivate and become the core of organizational project, and under the premise that living environment similar to cells in vivo is provided, how to carry out inducing cell and to break up towards direction initialization by controlling environmental stimuli or medicine controlled releasing, most important for cell, the dimensional culture of tissue and functionalization。In recent years, electronic stimulation development is very rapid, develops into adjustment differentiation of stem cells, startup limbs and spinal cord regeneration, induced ectopic eyeball from single electrical stimulation solution muscular movement problem and produces and control tumor growth etc.。
By external electricity irritation cell culture test device, it is possible to test out electrical stimulation signal and the speed of growth of cell, regeneration capacity, to the effect of specific direction differentiation and relation, thus providing experimental basis for electronic stimulation, improve the effect of electronic stimulation。But existing external electricity irritation cell culture test device, the electrical stimulation signal provided is usually the unidirectional current of voltage constant, also medicine cannot be added in culture environment, thus cannot to the ac signal of various frequencies and amplitude and the speed of growth of cell, regeneration capacity, study to effect and the relation of specific direction differentiation and test；More can not to ac signal and medicine to the speed of growth of cell, regeneration capacity, test to collaborative, inhibition and the relation of specific direction differentiation and study。Hinder the further development of electronic stimulation。
Summary of the invention
The first object of the present invention is to provide a kind of high-flux electric stimulates the experimental provision of Cell differentiation inducing activity and medicine controlled releasing, this device both can produce the electrical stimulation signal of wide range of frequencies and amplitude, medicine can also be simultaneously introduced, thus the high-throughout electrical stimulation signal carried out in stem cell differentiation in vitro process and the screening of medicine, provide experimental basis more extensive, reliable for stimulation therapy。
The present invention realizes the experimental provision that be the technical scheme is that a kind of high-flux electric stimulation Cell differentiation inducing activity and medicine controlled releasing of above-mentioned first order, including electrical stimulation signal generator and cell culture apparatus, it is characterised in that:
The composition of described electrical stimulation signal generator is: the general end of single-chip minimum system is connected with the input of digital to analog converter, and the outfan of digital to analog converter is connected with the input of Signal-regulated kinase；The outfan of Signal-regulated kinase is connected with the moved end of single-pole double-throw switch (SPDT) K；
Upper cover plate is stamped at the top of described cell culture apparatus, upper cover plate is embedded with upper cover plate and connects electric wire, the titanium alloy electrode of bottom and tabular that one end of upper cover plate connection electric wire passes upper cover plate is connected, and titanium alloy electrode is connected in the inner chamber that electric wire hangs on cell culture apparatus by upper cover plate；Described upper cover plate connect the other end of electric wire pass the first of the sidepiece of upper cover plate and single-pole double-throw switch (SPDT) K not moved end be connected；
The base plate of described cell culture apparatus is drug release plate, and described drug release plate is titanium alloy sheet；Drug release plate is 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 of single-pole double-throw switch (SPDT) K not moved end be connected；The other end of drug release plate ground wire is connected with the ground end of Signal-regulated kinase；The upper surface of described drug release plate is placed with medicine carrying conductive polymer film。
The second object of the present invention is to provide a kind of experimental provision using above-mentioned high-flux electric to stimulate Cell differentiation inducing activity and medicine controlled releasing and carries out the experimental technique of high-flux electric stimulation Cell differentiation inducing activity and medicine controlled releasing, the method can produce the electrical stimulation signal of optional frequency and amplitude when experiment, medicine can also be simultaneously introduced, thus the screening of the high-throughout electrical stimulation signal carried out in stem cell differentiation in vitro process and medicine, thus provide experimental basis more extensive, reliable for stimulation therapy。
The present invention realizes being employed technical scheme comprise that of above-mentioned second order, a kind of high-flux electric used described in claim 1 stimulates the experimental provision of Cell differentiation inducing activity and medicine controlled releasing to carry out high-flux electric and stimulates the experimental technique of Cell differentiation inducing activity and medicine controlled releasing, and its operation is:
A, cell are inoculated: be seeded in by stem cell on the medicine carrying conductive polymer film on drug release plate, and add culture fluid to cell culture apparatus, make culture fluid flood titanium alloy electrode；
B, signal occur: open electrical stimulation signal generator, make electrical stimulation signal generator produce the electrical stimulation signal of setpoint frequency and the square wave of amplitude, sine wave or sawtooth waveforms, or make the electrical stimulation signal of the direct current of electrical stimulation signal generator generation setting voltage value；
C, electricity irritation medicine controlled releasing: operation single-pole double-throw switch (SPDT) make its moved end with first not moved end connect, the electrical stimulation signal of electrical stimulation signal generator generation passes sequentially through titanium alloy electrode, culture fluid, medicine carrying conductive polymer film and drug release plate and returns to electrical stimulation signal generator；Medicine in medicine carrying conductive polymer film, is discharged in culture fluid, thus carrying out electricity irritation medicine controlled releasing under the stimulation of electrical stimulation signal controls；
D, electricity irritation cell differentiation: when the electricity irritation of C step carried out to the time set, operation single-pole double-throw switch (SPDT) make its moved end with second not moved end connect, electrical stimulation signal generator generation electrical stimulation signal return to electrical stimulation signal generator by medicine carrying conductive polymer film and drug release plate；The stem cell being inoculated on medicine carrying conductive polymer film is subject to the stimulation of electrical stimulation signal, induction produces cell differentiation。
E, end: after the electricity irritation cell differentiation of D step reaches the time set, closing electrical stimulation signal generator, taken out by the stem cell in cell culture apparatus, namely experiment terminates。
Work process and the principle of the electrical stimulation signal generator of the present invention be:
The digital signal corresponding with the unidirectional current of setpoint frequency and the square wave of amplitude, sine wave, sawtooth waveforms or setting voltage value is produced by single-chip minimum system, this digital signal is converted to analogue signal by digital to analog converter, then through the galvanic electrical stimulation signal that Signal-regulated kinase conditioning is setpoint frequency and the square wave of amplitude, sine wave, sawtooth waveforms or setting voltage value, flow to cell culture apparatus finally by single-pole double-throw switch (SPDT) K。
Compared with prior art, the beneficial effects of the present invention is:
Assembly of the invention and method can accurately control to produce the electrical stimulation signal of variety classes, amplitude and frequency, can when identical temperature, humidity and media environment, the voltage/current stimulus signal of research variety classes, amplitude and frequency and the relation of cell differentiation and medicine controlled releasing, thus the high-throughout electrical stimulation signal carried out in stem cell differentiation in vitro process and the screening of medicine, provide experimental basis more extensive, reliable for stimulation therapy。
Further, the cell culture apparatus of the present invention is 1～200。
The present invention can once be synchronously performed high flux Cell differentiation inducing activity and the medicine controlled releasing experiment of multi-medicament and stem cell, more efficiently can quickly filter out the medicine most beneficial for cell differentiation and electrical stimulation signal。
Below in conjunction with the drawings and specific embodiments, the present invention is described in further details:
Accompanying drawing explanation
Fig. 1 is the overall structure schematic diagram of apparatus of the present invention。
The circuit theory diagrams of Fig. 2 electrical stimulation signal generator。
Detailed description of the invention
Fig. 1-2 illustrates, a kind of detailed description of the invention of the present invention is, 1. the experimental provision of a 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 described electrical stimulation signal generator 1 is: the general end of single-chip minimum system MCU is connected with the input of digital to analog converter (DAC), and the outfan of digital to analog converter (DAC) is connected with the input of Signal-regulated kinase SP；The outfan of Signal-regulated kinase 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 described cell culture apparatus 7, upper cover plate 6 is embedded with upper cover plate and connects electric wire (4), the titanium alloy electrode 8 of bottom and tabular that one end of upper cover plate connection electric wire (4) passes upper cover plate 6 is connected, and titanium alloy electrode 8 is connected in the inner chamber that electric wire (4) hangs on cell culture apparatus 7 by upper cover plate；Described upper cover plate connect the other end of electric wire (4) pass the first of the sidepiece of upper cover plate 6 and single-pole double-throw switch (SPDT) K not moved end (Ka) be connected；
The base plate of described cell culture apparatus 7 is drug release plate 9, and described drug release plate 9 is titanium alloy sheet；Drug release plate 9 is welded with drug release switching plate connecting line 5 and drug release plate ground wire 3, the other end of drug release switching plate connecting line 5 with the second of single-pole double-throw switch (SPDT) K not moved end Kb be connected；The other end of drug release plate ground wire 3 is connected with the ground end of Signal-regulated kinase SP；The upper surface of described drug release plate 9 is placed with medicine carrying conductive polymer film 11。
The cell culture apparatus 7 of this example is 1～200。
A kind of high-flux electric making this example stimulates the experimental provision of Cell differentiation inducing activity and medicine controlled releasing to carry out high-flux electric and stimulates the experimental technique of Cell differentiation inducing activity and medicine controlled releasing, and its operation is:
A, cell are inoculated: are seeded in by stem cell on the medicine carrying conductive polymer film 11 on drug release plate 9, and add culture fluid to cell culture apparatus 7, make culture fluid flood titanium alloy electrode 8；
B, signal occur: open electrical stimulation signal generator 1, make electrical stimulation signal generator (1) produce the electrical stimulation signal of setpoint frequency and the square wave of amplitude, sine wave or sawtooth waveforms, or make the electrical stimulation signal of the direct current of electrical stimulation signal generator generation setting voltage value；
C, electricity irritation medicine controlled releasing: operation single-pole double-throw switch (SPDT) makes its moved end Kc and the first not moved end (Ka) connect, and the electrical stimulation signal that electrical stimulation signal generator 1 produces passes sequentially through titanium alloy electrode 8, culture fluid, medicine carrying conductive polymer film 11 and drug release plate 9 and returns to electrical stimulation signal generator 1；Medicine in medicine carrying conductive polymer film 11, is discharged in culture fluid, thus carrying out electricity irritation medicine controlled releasing under the stimulation of electrical stimulation signal controls；
D, electricity irritation cell differentiation: when the electricity irritation of C step carried out to the time set, operation single-pole double-throw switch (SPDT) makes its moved end Kc and the second not moved end (Kb) connect, and the electrical stimulation signal that electrical stimulation signal generator 1 produces returns to electrical stimulation signal generator 1 by medicine carrying conductive polymer film 11 and drug release plate 9；The stem cell being inoculated on medicine carrying conductive polymer film 11 is subject to the stimulation of electrical stimulation signal, induction produces cell differentiation。
E, end: after the electricity irritation cell differentiation of D step reaches the time set, closing electrical stimulation signal generator 1, taken out by the stem cell in cell culture apparatus 7, namely experiment terminates。
Claims (3)
1. high-flux electric stimulates an experimental provision for 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 described electrical stimulation signal generator (1) is: the general end of single-chip minimum system (MCU) is connected with the input of digital to analog converter (DAC), and the outfan of digital to analog converter (DAC) is connected with the input of Signal-regulated kinase (SP)；The outfan of Signal-regulated kinase (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 described cell culture apparatus (7), upper cover plate (6) is embedded with upper cover plate and connects electric wire (4), the titanium alloy electrode (8) of bottom and tabular that one end of upper cover plate connection electric wire (4) passes upper cover plate (6) is connected, and titanium alloy electrode (8) is connected in the inner chamber that electric wire (4) hangs on cell culture apparatus (7) by upper cover plate；Described upper cover plate connect the other end of electric wire (4) pass the first of the sidepiece of upper cover plate (6) and single-pole double-throw switch (SPDT) (K) not moved end (Ka) be connected；
The base plate of described cell culture apparatus (7) is drug release plate (9), and described drug release plate (9) is titanium alloy sheet；Drug release plate (9) is welded with drug release switching plate connecting line (5) and drug release plate ground wire (3), and the second of the other end of drug release switching plate connecting line (5) and single-pole double-throw switch (SPDT) (K) not moved end (Kb) is connected；The other end of drug release plate ground wire (3) is connected with the ground end of Signal-regulated kinase (SP)；The upper surface of described drug release plate (9) is placed with medicine carrying conductive polymer film (11)。
2. high-flux electric according to claim 1 stimulates the experimental provision of Cell differentiation inducing activity and medicine controlled releasing, it is characterised in that: described cell culture apparatus (7) is 1～200。
3. using the high-flux electric described in claim 1 to stimulate the experimental provision of Cell differentiation inducing activity and medicine controlled releasing to carry out high-flux electric and stimulate an experimental technique for Cell differentiation inducing activity and medicine controlled releasing, its operation is:
A, cell are inoculated: be seeded in by stem cell on the medicine carrying conductive polymer film (11) on drug release plate (9), and add culture fluid to cell culture apparatus (7), make culture fluid flood titanium alloy electrode (8)；
B, signal occur: open electrical stimulation signal generator (1), make electrical stimulation signal generator (1) produce the electrical stimulation signal of setpoint frequency and the square wave of amplitude, sine wave or sawtooth waveforms, or make the electrical stimulation signal of the direct current of electrical stimulation signal generator generation setting voltage value；
C, electricity irritation medicine controlled releasing: operation single-pole double-throw switch (SPDT) (K) make its moved end (Kc) with first not moved end (Ka) connect, the electrical stimulation signal that electrical stimulation signal generator (1) produces passes sequentially through titanium alloy electrode (8), culture fluid, medicine carrying conductive polymer film (11) and drug release plate (9) and returns to electrical stimulation signal generator (1)；Medicine in medicine carrying conductive polymer film (11), is discharged in culture fluid, thus carrying out electricity irritation medicine controlled releasing under the stimulation of electrical stimulation signal controls；
D, electricity irritation cell differentiation: when the electricity irritation of C step carried out to the time set, operation single-pole double-throw switch (SPDT) make its moved end (Kc) with second not moved end (Kb) connect, the electrical stimulation signal that electrical stimulation signal generator (1) produces passes through medicine carrying conductive polymer film (11) and drug release plate (9) and returns to electrical stimulation signal generator (1)；The stem cell being inoculated on medicine carrying conductive polymer film (11) is subject to the stimulation of electrical stimulation signal, induction produces cell differentiation。
E, end: after the electricity irritation cell differentiation of D step reaches the time set, closing electrical stimulation signal generator (1), taken out by the stem cell in cell culture apparatus (7), namely 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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| CN105695329B CN105695329B (en) | 2018-08-17 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114621947A (en) * | 2022-03-30 | 2022-06-14 | 中国科学院深圳先进技术研究院 | Stem cell neural differentiation regulation and control technology based on electrical stimulation |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114621947A (en) * | 2022-03-30 | 2022-06-14 | 中国科学院深圳先进技术研究院 | Stem cell neural differentiation regulation and control technology based on electrical stimulation |
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