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WO2013128630A1 - Récipient de culture de cellules, dispositif de culture de cellules l'utilisant, et procédé de culture de cellules - Google Patents

Récipient de culture de cellules, dispositif de culture de cellules l'utilisant, et procédé de culture de cellules Download PDF

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Publication number
WO2013128630A1
WO2013128630A1 PCT/JP2012/055372 JP2012055372W WO2013128630A1 WO 2013128630 A1 WO2013128630 A1 WO 2013128630A1 JP 2012055372 W JP2012055372 W JP 2012055372W WO 2013128630 A1 WO2013128630 A1 WO 2013128630A1
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WIPO (PCT)
Prior art keywords
cell culture
cell
electrode
culture container
spheroid
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Ceased
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PCT/JP2012/055372
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English (en)
Japanese (ja)
Inventor
広斌 周
亮介 高橋
明子 久田
浩 園田
志津 松岡
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Hitachi Ltd
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Hitachi Ltd
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Priority to JP2014501927A priority Critical patent/JP5903156B2/ja
Priority to PCT/JP2012/055372 priority patent/WO2013128630A1/fr
Publication of WO2013128630A1 publication Critical patent/WO2013128630A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • 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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/06—Plates; Walls; Drawers; Multilayer plates
    • C12M25/08—Plates; Walls; Drawers; Multilayer plates electrically charged
    • 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
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0062—General methods for three-dimensional culture

Definitions

  • the present invention relates to a technique for culturing animal cells using a culture vessel and forming a cell spheroid of a spherical tissue (three-dimensional tissue) of the cell.
  • the cells to be cultured are mainly parenchymal cells that are responsible for various organ functions, but it is generally difficult to perform long-term culture in a state where the functions of the parenchymal cells are maintained in vitro.
  • Various attempts have been made to solve the problem.
  • the spheroid culture system which is cultured in a state where a large number of cells are aggregated three-dimensionally, is an excellent in vitro cell culture system from the viewpoint of maintaining cell polarity and cell-cell interaction. is there.
  • Many reports that have been applied to many types of cells such as osteoblasts and hepatocytes.
  • spheroid culture of hepatocytes has a possibility of being applied to an artificial liver and a possibility of being applied to a drug metabolism simulator in vitro.
  • cell spheroid culture is an excellent parenchymal cell culture system in vitro, but as a disadvantage, a method for spheroidizing without damaging cells and a simple method for maintaining spheroidized cells have not been established Can be mentioned.
  • Non-patent Document 1 a method of forcibly keeping cells in a floating state by physical force (eg, stirring of a culture solution) and increasing the adhesion probability between cells
  • Non-patent Document 2 There is a method (Non-patent Document 2) that promotes spheroidization by modifying the surface of the cell culture or the shape of the culture vessel.
  • the cell spheroids formed by the method described in the prior art have a problem that they are highly peelable from the equipment and are lost during the medium exchange process, and the size cannot be controlled and it is difficult to maintain the quality. Furthermore, it has the subject that the formation of the cell spheroid by the self-organization of a cell takes time, and the handleability of the formed cell spheroid is low.
  • an object of the present invention is to provide a cell culture container that can form a cell spheroid of a three-dimensional cell tissue uniformly and efficiently using dielectrophoresis effective for cell manipulation.
  • the present invention employs, for example, the configuration described in the claims in order to solve the above problems.
  • a bottom substrate, a side substrate, and a top substrate that form a cell culture space, and an induction mechanism that induces cells in the cell culture space to a cell spheroid formation region by a dielectrophoretic force.
  • the induction mechanism has an electrode that generates a non-uniform electric field in the cell culture space.
  • the guide mechanism having the electrodes may be composed of a plurality of pairs of electrodes provided in the circumferential direction of the side substrate.
  • the guiding mechanism having the electrode comprises a conductive film-forming electrode formed on the surface of a plurality of protrusions provided on the bottom substrate and an electrode provided on the top substrate. Good.
  • the cell culture device of the present invention comprises the above-described cell culture container and an AC power source that applies AC to the electrode of the induction mechanism.
  • the cell culture method of the present invention induces cells in a cell culture space to a cell spheroid formation region by dielectrophoretic force by applying an alternating current to the electrode using the above cell culture container. .
  • the present invention it is possible to uniformly and efficiently form a cell spheroid of a cell three-dimensional tissue. Furthermore, the formation of an extracellular matrix can be promoted by the ion-inducing effect of electric lines of force, and cell spheroids can be formed more quickly.
  • the dielectrophoretic force F DEP acting on the dielectric particles is given by the following formula 1 (Non-patent Document 3).
  • Non-patent Document 3 a case where the dielectric particles are cells will be described as an example.
  • a cell radius when approximated to a sphere
  • ⁇ 0 dielectric constant of vacuum
  • ⁇ m relative dielectric constant of medium
  • E electric field strength
  • ⁇ is an operator representing a gradient.
  • ⁇ E 2 is the gradient of the square of the electric field strength E 2 , so it means how much E 2 has an inclination at that position, that is, how much the electric field changes spatially.
  • K is called the Clausius Mosotti number and is expressed by Equation 2.
  • ⁇ b * and ⁇ m * represent the complex dielectric constant of the cell and the medium, respectively.
  • Re [K] is defined to represent the real part of the Clausius Mosotti number
  • Re [K]> 0 represents positive dielectrophoresis, and the cells migrate in the same direction as the electric field gradient, that is, toward the electric field concentration part. Is done.
  • Re [K] ⁇ 0 represents negative dielectrophoresis and migrates in the direction away from the electric field concentration portion, that is, toward the weak electric field portion.
  • Equation 3 the complex dielectric constant ⁇ r * is expressed by Equation 3.
  • ⁇ r represents the relative dielectric constant of the cell or the medium
  • ⁇ represents the conductivity of the cell or the medium
  • ⁇ represents the angular frequency of the applied electric field.
  • FIG. 1 is a plan view of a cell culture container
  • FIG. 2 is a cross-sectional view taken along line A-A ′ of FIG.
  • 1 is the ceiling substrate of the cell culture container
  • 2 is the bottom substrate of the cell culture container
  • 3 is the side substrate of the cell culture container.
  • Reference numerals 5A, 5B, 6A, and 6B denote electrodes for inducing cells provided on the side substrate of the cell culture container.
  • 4A and 4B are electrodes for fixing the cell aggregate.
  • Reference numeral 8 denotes an internal space of the culture container, which is filled with a medium containing cells 7.
  • Reference numerals 9 and 10 are AC power supplies, and 11 is an impedance measuring device for measuring the impedance between the electrode pairs 4A and 4B.
  • 12A indicates the electric lines of force between the electrodes 5A, 5B, 6A, and 6B
  • 12B indicates the electric lines of force between the electrodes 4A and 4B.
  • the above-described ceiling substrate 1 and bottom substrate 2 can be formed using glass, silicon, quartz, or an insulating solid substrate such as plastics and polymers as a base material. More preferably, the bottom substrate 2 is made of a material that can be surface-modified by pretreatment so as not to allow cells to adhere to the surface. It is desirable to provide.
  • a material of any one of platinum, gold, chromium, palladium, rhodium, silver, aluminum, tungsten, ITO, or a combination thereof can be used.
  • FIG. 3 is a diagram showing the relationship between the frequency of the AC electric field and the real part Re [K] of the Clausius-Mottie number. It can be seen that when the conductivity of the medium is 1000 mS / m or more, the dielectrophoresis is all negative dielectrophoresis within a frequency of 10 9 Hz. That is, the cells migrate toward the direction away from the electric field concentration portion, that is, toward the weak electric field portion. Since the dielectrophoretic force is proportional to the magnitude of Re [K], the applied frequency is preferably 100 to 10 MHz.
  • the cell induction mechanism electrode of this example has a weak electric field portion formed at the center of the four electrodes 5A, 5B, 6A, and 6B. Therefore, the cells in the high conductivity medium can move to the weak electric field and aggregate.
  • the circular shape has been described as the shape of the induction mechanism electrode, but it is needless to say that the shape may be a quadrangle or a polygon. Moreover, although the number of electrodes was demonstrated with four, four or more even-numbered electrodes may be sufficient.
  • the present invention is not limited to the above-described embodiment. If the electrode shape forms a weak electric field part as shown in FIG. 1, the cells in the high conductivity medium are made into the weak electric field part by negative dielectrophoretic force. Can be moved or fixed. Furthermore, in order to control the shape of the cell spheroid, multilayer induction electrodes 5A1, 5A2, 5A3, 6A1, 6A2, 6A3, 5B1, 5B2, and 5B3 as shown in FIG. 4 may be provided. In this cell culture container, different AC electric fields are applied to the electrodes of each layer, and the shape of the cell aggregate can be controlled by dielectrophoretic forces having different strengths generated in each layer.
  • the upper and lower electrodes 5A1, 5A3, 6A1, 6A3, 5B1, and 5B3 are moved from the upper and lower sides to the central portion.
  • Cells can be moved to create spherical cell aggregates.
  • Cells 7 are seeded in the culture space 8 of the culture vessel.
  • an AC electric field is first applied to the cell induction electrodes 5A, 5B, 6A, and 6B from the AC power source 9 in FIG. 1 under the conditions of 5% CO 2 and 37 ° C. using a CO 2 incubator.
  • the cells rapidly gather at the center of the culture vessel due to the negative dielectrophoretic force and become cell aggregates.
  • an intercellular adhesion structure or an extracellular matrix is formed, and a cell spheroid is formed.
  • the AC power source 9 is turned off. Medium exchange is performed from here.
  • an AC electric field is applied from the AC power supply 10 to the electrodes 4A and 4B in order to fix the cell spheroids during medium exchange.
  • the cell spheroid fixed to the electrode 4B side by the negative dielectrophoretic force is not flowed at the time of medium exchange and is held in the culture vessel.
  • a spherical three-dimensional tissue having a uniform size is formed by stationary culture for a certain period. This is an effective culture method for cell assays and the like because it is considered that the activity of cells originally retained is not lost.
  • the electric lines of force 12A and 12B have an effect of promoting the movement of intercellular ions, it is possible to promote the formation of the extracellular matrix by periodically applying to the cell spheroids.
  • the number of cells induced in the weak electric field part, the size of the spheroid, and the like can be estimated.
  • a method for estimating the size and thickness of the cell spheroid by measuring the impedance between the fixed mechanism electrodes 4A and 4B will be described. A description will be given using FIG. 5 and Equations 4 to 8, assuming that the interelectrode impedance is Z, the capacitance is C, the reactance is x, the resistance is r, and the resistance is R.
  • Equation 4 represents the combined impedance Z of the CR parallel equivalent circuit
  • Equation 5 represents the resistance r of the CR parallel equivalent circuit
  • Equation 6 represents the reactance x of the CR parallel equivalent circuit
  • Equation 7 represents the CR parallel equivalent circuit
  • Equation 8 representing the resistance R
  • Equation 8 represents the capacitance C of the CR parallel equivalent circuit.
  • FIG. 5 shows an electrical circuit between the electrodes 13 of the cell culture container with an equivalent circuit.
  • a medium containing cells is present between the electrodes 13.
  • the capacitance C14 configured with the culture medium as an interelectrode dielectric and the electric conduction resistance R15 due to the culture medium connect the electrodes 13 in parallel. That is, the number of local cells, the size of cell aggregates, and the like can be estimated from the amount of change in impedance between the upper and lower electrodes 4A and 4B of the culture vessel. As a result, the state of cell spheroid formation can be evaluated simply and quickly using an electrical signal instead of observation with an optical microscope.
  • a culture plate for forming a large amount of spheroids is composed of a cell culture plate hole 17 and a frame 16 for collecting the cell culture containers as shown in FIG.
  • the main culture plate can simultaneously form a large number of cell spheroids and is suitable for comparative experiments such as toxicity.
  • the induction electrode and the fixed electrode of the culture plate can be produced by a semiconductor fine wiring technique or the like.
  • uniform size cell spheroids can be produced in a shorter period of time by using the cell culture container of the present invention.
  • Preparation of hepatocytes follows the in situ collagenase flow-through method. Details are as follows. Fisher 344 male rats (7-10 weeks old) are laparotomized under pentobarbital anesthesia, a catheter is inserted into the portal vein, and a pre-perfusion solution (Ca2 + and Mg2 + -free Hanks solution containing ETCA) is injected. The liver is cut off, minced in chilled Hanks solution, and dispersed to cells by pipetting. Subsequently, undigested tissue is removed by gauze filtration. The cell suspension removes non-parenchymal cells by repeating centrifugation at 50G for 1 minute several times.
  • a pre-perfusion solution Ca2 + and Mg2 + -free Hanks solution containing ETCA
  • injured hepatocytes are removed by centrifugation at 500 G for 5 minutes.
  • the survival rate of the obtained hepatocytes is measured by trypan blue exclusion method, and hepatocytes having a survival rate of 85% or more are used for culture.
  • hepatocytes having a survival rate of 85% or more are used for the culture, but it is needless to say that the conditions are not necessarily limited thereto.
  • the prepared hepatocytes are seeded in a culture vessel at a seeding concentration of 1 ⁇ 10 5 cells / ml.
  • an AC electric field having a frequency of 100 kHz and a voltage of 20 Vpp is first applied to the cell induction electrodes 5A, 5B, 6A, and 6B from the AC power source 9 in FIG. 1 under the conditions of 5% CO 2 and 37 ° C. using a CO 2 incubator. Apply. As a result, hepatocytes gather in the center of the culture vessel due to the negative dielectrophoretic force and become hepatocyte aggregates. Next, by culturing for 12 hours as it was, an adhesion structure between hepatocytes was formed and a hepatocyte spheroid was formed. At this time, the AC power source 9 is turned off, and the medium is changed every 24 hours.
  • an AC electric field having a frequency of 100 kHz and a voltage of 20 Vpp is applied from the AC power source 10 to the electrodes 4A and 4B.
  • the hepatocyte spheroids fixed by the negative dielectrophoretic force are retained in the culture vessel without being washed away during the medium exchange.
  • a spherical three-dimensional tissue having a uniform size was formed. This is an effective culture method for cell assays and the like because it is considered that the activity of cells originally retained is not lost.
  • This example shows an example using rat hepatocytes, but it can be applied to various animal and plant cell types as described above, and the cell types are not particularly limited.
  • This example is a cell culture container having a cell-adhesive bottom substrate electrode 4B.
  • the cell induction electrodes 5A, 5B, 6A, and 6B for inducing cells are the same as in FIG. This example will be described with reference to FIG.
  • the surface of the electrode 4B is subjected to a surface treatment for cell adhesion.
  • Cells 7 are seeded in the culture space 8 of the culture vessel.
  • an AC electric field is first applied to the cell induction electrodes 5A, 5B, 6A, and 6B from the AC power source 9 in FIG. 1 under the conditions of 5% CO 2 and 37 ° C. using a CO 2 incubator.
  • the cells rapidly gather at the center of the culture vessel due to the negative dielectrophoretic force and become cell aggregates. Since the surface of the bottom substrate electrode 4B has cell adhesiveness, the cell aggregate can be fixed.
  • an intercellular adhesion structure or an extracellular matrix is formed, and a cell spheroid is formed.
  • the AC power source 9 is turned off. Medium exchange is performed from here.
  • the cell spheroid is not flowed because it is fixed to the surface of the bottom substrate electrode.
  • a spherical three-dimensional tissue having a uniform size is formed by stationary culture for a certain period.
  • a DC electric field is applied between the DC power source 18 and the electrodes 4A and 4B, and the cell spheroids are peeled from the culture surface 4B due to the effect of electrolysis of the medium occurring on the electrode 4B surface. be able to.
  • Cell spheroids can be detached without using an enzyme (for example, trypsin) used in conventional cell detachment. Thereby, cell damage such as proteolysis can be avoided.
  • uniform size cell spheroids can be produced in a shorter period of time by using the cell culture container of the present invention.
  • a culture sheet having a nanopillar structure is provided on the bottom substrate of the culture vessel.
  • 8 and 9 show the cross-sectional structure of the culture vessel of this example.
  • a culture sheet having a nanopillar structure 19 is provided on the culture vessel bottom substrate 2, a conductive film 20 is deposited on the surface, and a counter electrode 21 is further provided.
  • a conductive film 20 is deposited on the surface, and a counter electrode 21 is further provided.
  • the tip of the nanopillar is in a strong electric field region, cell spheroids are formed with a positive dielectrophoretic force. For this reason, it is necessary to lower the conductivity of the culture medium below the region where positive dielectrophoresis works (see FIG. 3). This conductivity adjustment can be performed by mixing a low conductivity sugar solution or the like in the medium.
  • an AC electric field is first applied to the electrodes 20 and 21 from the AC power source 10 in FIG. 8 under the conditions of 5% CO 2 and 37 ° C. using a CO 2 incubator.
  • the cells rapidly gather on the nanopillar 19 which is a strong electric field region and become an aggregate of cells by a positive dielectrophoretic force. Since the surface of the nanopillar 19 has cell adhesiveness, cell aggregates can be fixed.
  • the switch 22A is turned OFF and the AC power supply 10 is turned off.
  • Cell spheroids are not flowed because they are fixed to the nanopillar structure. Thereafter, a spherical three-dimensional tissue having a uniform size is formed by stationary culture for a certain period.
  • the switch 22B is turned on, a DC electric field is applied between the electrodes 20 and 21 from the DC power source 18, and the cell spheroids are removed by the effect of electrolysis of the medium occurring on the electrode 20 surface. It can peel from the nanopillar 19.
  • the impedance between the electrodes 20 and 21 can be measured using the impedance measuring device 11, and the number of cells, the size of the cell spheroid, and the like can be estimated from the amount of change in the impedance between the electrodes 20 and 21 of the culture vessel.
  • FIG. 9 is a modified version of the nanopillar structure of FIG. 8, and the pillar at the center of the nanopillar structure of FIG. 8 is higher than the surrounding pillars.
  • the electric field concentrates on the high pillar at the center, and the cells rapidly gather on the nanopillar 19 of the culture vessel by the positive dielectrophoretic force to form cell aggregates. Is possible. Further, the cell spheroid formed with this structure is located at the center of the nanopillar sheet, and can be easily observed and measured.
  • the experimental conditions are the same as in the experiment of Example 1.
  • the conductivity of the rat hepatocyte suspension was adjusted to 70 mS / m or less, and seeded at a seeding concentration of 1 ⁇ 10 5 cells / ml in the cell culture container of Example 3.
  • an AC electric field having a frequency of 100 kHz and a voltage of 20 Vpp is first applied to the cell induction electrodes 20 and 21 from the AC power source 10 in FIG. 8 under the conditions of 5% CO 2 and 37 ° C. using a CO 2 incubator.
  • hepatocytes gather on the nanopillar 19 having cell adhesion formed in the center of the culture vessel by the positive dielectrophoretic force and become hepatocyte aggregates.
  • hepatocyte spheroids were formed by forming an adhesion structure between hepatocytes.
  • the AC power supply 10 is turned off.
  • the medium is changed every 24 hours from here. Since the hepatocyte spheroids are adhered on the nanopillars having adhesiveness, they are not detached at the time of medium exchange and are maintained in the culture vessel.
  • the size and thickness of the hepatocyte spheroid can be estimated by measuring the impedance change between the electrodes by the impedance measuring device 11 during culture. From the static culture for 72 hours, a spherical three-dimensional tissue having a uniform size was formed. After culturing, a 1 V voltage is applied from the DC power source 18 to the electrodes 20 and 21, and the hepatocyte spheroid can be peeled off from the nanopillar.
  • the present invention is not limited to the above embodiments as long as the characteristics of the present invention are not impaired, and other forms conceivable within the scope of the technical idea of the present invention are also included in the scope of the present invention.

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PCT/JP2012/055372 2012-03-02 2012-03-02 Récipient de culture de cellules, dispositif de culture de cellules l'utilisant, et procédé de culture de cellules Ceased WO2013128630A1 (fr)

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JP2014501927A JP5903156B2 (ja) 2012-03-02 2012-03-02 細胞培養容器、それを用いた細胞培養装置および細胞培養方法
PCT/JP2012/055372 WO2013128630A1 (fr) 2012-03-02 2012-03-02 Récipient de culture de cellules, dispositif de culture de cellules l'utilisant, et procédé de culture de cellules

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017033898A1 (fr) * 2015-08-25 2017-03-02 旭硝子株式会社 Dispositif de culture de cellules et procédé de production d'un échantillon biologique
CN109055195A (zh) * 2018-03-26 2018-12-21 宁波华仪宁创智能科技有限公司 非接触式胚胎移动装置及方法
KR20190048336A (ko) * 2017-10-31 2019-05-09 한양디지텍 주식회사 세포 및 배양액 조건에 따른 전기자극 변수 제어장치 및 방법
WO2022109319A1 (fr) * 2020-11-20 2022-05-27 The Regents Of The University Of California Nouveaux systèmes de culture tissulaire et procédé de culture par gravité réduite pour la production de tissu vascularisé

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WO2008018390A1 (fr) * 2006-08-10 2008-02-14 Tohoku University Procédé de formation d'un motif de cellules
WO2010079602A1 (fr) * 2009-01-08 2010-07-15 株式会社日立製作所 Procédé pour la culture d'hépatocytes animaux
WO2010150521A1 (fr) * 2009-06-23 2010-12-29 株式会社日立製作所 Substrat de culture, feuille de culture, et procédé de culture cellulaire

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Publication number Priority date Publication date Assignee Title
WO2008018390A1 (fr) * 2006-08-10 2008-02-14 Tohoku University Procédé de formation d'un motif de cellules
WO2010079602A1 (fr) * 2009-01-08 2010-07-15 株式会社日立製作所 Procédé pour la culture d'hépatocytes animaux
WO2010150521A1 (fr) * 2009-06-23 2010-12-29 株式会社日立製作所 Substrat de culture, feuille de culture, et procédé de culture cellulaire

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LIN R-Z. ET AL.: "Recent advances in three-dimensional multicellular spheroid culture for biomedical research", BIOTECHNOL. J., vol. 3, 2008, pages 1172 - 1184, XP002558418, DOI: doi:10.1002/biot.200700228 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017033898A1 (fr) * 2015-08-25 2017-03-02 旭硝子株式会社 Dispositif de culture de cellules et procédé de production d'un échantillon biologique
KR20190048336A (ko) * 2017-10-31 2019-05-09 한양디지텍 주식회사 세포 및 배양액 조건에 따른 전기자극 변수 제어장치 및 방법
KR102499114B1 (ko) * 2017-10-31 2023-02-13 주식회사 유스바이오글로벌 세포 및 배양액 조건에 따른 전기자극 변수 제어장치 및 방법
CN109055195A (zh) * 2018-03-26 2018-12-21 宁波华仪宁创智能科技有限公司 非接触式胚胎移动装置及方法
CN109055195B (zh) * 2018-03-26 2023-09-05 广州市华粤行医疗科技有限公司 非接触式胚胎移动装置及方法
WO2022109319A1 (fr) * 2020-11-20 2022-05-27 The Regents Of The University Of California Nouveaux systèmes de culture tissulaire et procédé de culture par gravité réduite pour la production de tissu vascularisé

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