WO2017047986A1 - Cell spheroid cultivator, and cell spheroid culture system containing same - Google Patents

Cell spheroid cultivator, and cell spheroid culture system containing same Download PDF

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Publication number
WO2017047986A1
WO2017047986A1 PCT/KR2016/010128 KR2016010128W WO2017047986A1 WO 2017047986 A1 WO2017047986 A1 WO 2017047986A1 KR 2016010128 W KR2016010128 W KR 2016010128W WO 2017047986 A1 WO2017047986 A1 WO 2017047986A1
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Prior art keywords
cell spheroid
cell
culture
incubator
spheroid
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PCT/KR2016/010128
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French (fr)
Korean (ko)
Inventor
정초록
정홍렬
김장환
김태돈
손명진
손미영
오수진
임정화
정경숙
조현수
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한국생명공학연구원
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Publication of WO2017047986A1 publication Critical patent/WO2017047986A1/en

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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
    • C12M23/08—Flask, bottle or test tube
    • 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
    • C12M23/38—Caps; Covers; Plugs; Pouring means
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis

Definitions

  • the present invention relates to a cell spheroid incubator and a cell spheroid culture system including the same, and more particularly, to perform cell spheroid culture more easily, to provide a cell spheroid with high yield, and to produce cell spheroid culture.
  • the present invention relates to a cell spheroid incubator and a cell spheroid culture system including the same, which can further lower the unit cost.
  • Cell culture refers to a process of culturing cells isolated from an organism. Aseptically select biological tissues, treat them with digestive enzymes such as trypsin or pronase, separate them into single cells, perform primary cultures, and transplant single cells obtained by dispersing passaged cell lines or cell lines with the same enzyme treatment to the growth medium. There is a method of dispersing and culturing single cells by treatment with proteolytic enzymes such as inoculation and subsequent passage.
  • the cell culture method is to culture monolayer cells in a two-dimensional flat container. This two-dimensional cell culture and modeling provides key insights into the mechanisms of cell motility and transmission and disease.
  • Three-dimensional cell culture methods in this regard include a hanging-drop method, a rotary culture method, a centrifugal separation method, a micro molding method, and related patents, such as Korean Patent Registration No. 10-1341572 (Modern Culture Method), Korea Patent No. 10-1412155 (rotary culture method), Korea Patent No. 10-1362293 (micro molding method) and the like.
  • the present invention has been made to solve the above problems, the problem of the present invention is to perform the cell spheroid culture more easily, to provide a cell spheroid with a high yield, the cell spheroid culture production cost more It is to provide a cell spheroid incubator and a cell spheroid culture system comprising the same.
  • Cell spheroid incubator has an opening, the body portion containing the cell culture solution, a mesh-shaped net is installed along the inner wall of the body portion, the filter portion is installed inside the body portion detachably from the body portion And is coupled to the opening of the body portion, and includes a cap portion formed with a through hole capable of supplying gas.
  • Cell spheroid culture system has an opening and the body portion containing the cell culture solution, including a mesh-shaped mesh is installed along the inner wall of the body portion and the filter portion is installed inside the body portion to be separated from the body portion,
  • a cell spheroid incubator coupled to an opening of the body part and including a cap part formed with a through hole capable of supplying gas;
  • a rotator having a mounting portion on which the cell spheroid incubator is mounted and rotating the cell spheroid incubator; And a chamber containing a rotator equipped with a cell spheroid incubator.
  • the cell spheroid incubator according to the present invention includes a body portion containing the cell culture solution, a mesh in the form of a mesh, a filter portion installed inside the body portion, and a cap portion coupled to the opening portion and having a through hole capable of supplying gas.
  • the cell spheroid culture system includes a chamber containing a cell spheroid incubator, a rotator for rotating the cell spheroid incubator, and a rotator equipped with the cell spheroid incubator, thereby making cell spheroid culture more convenient.
  • a chamber containing a cell spheroid incubator a rotator for rotating the cell spheroid incubator, and a rotator equipped with the cell spheroid incubator, thereby making cell spheroid culture more convenient.
  • Example 1 is a perspective view showing a cell spheroid incubator according to Example 1 of the present invention.
  • Figure 2 is a perspective view and a partial cross-sectional view for showing the internal structure of the cell spheroid incubator according to Example 1 of the present invention.
  • Figure 3 is a plan view showing a cell spheroid incubator according to Example 1 of the present invention.
  • Figure 4 is a partial cross-sectional view for showing the cap portion of the cell spheroid incubator according to Example 1 of the present invention.
  • Example 5 is a perspective view showing a rotator of a cell spheroid culture system according to Example 2 of the present invention.
  • FIG. 6 is a perspective view showing a chamber of the cell spheroid culture system according to Example 2 of the present invention.
  • Example 7 is a perspective view showing a cell spheroid culture system according to Example 2 of the present invention.
  • Example 8 is a view showing the appearance of the 293 cell spheroid produced through the cell spheroid culture system according to Example 2 of the present invention.
  • FIG. 9 is a view showing the appearance of various other cell spheroids produced through the cell spheroid culture system according to Example 2 of the present invention.
  • FIG 10 is a front view showing the cell spheroid culture system according to Example 3 of the present invention.
  • Example 12 is a right side view showing the cell spheroid culture system according to Example 3 of the present invention.
  • Figure 13 is a conceptual diagram showing the operating mechanism of the cell spheroid culture system according to Example 3 of the present invention.
  • FIG. 1 is a perspective view showing a cell spheroid incubator according to Example 1 of the present invention.
  • Figure 2 is a perspective view and a partial cross-sectional view for showing the internal structure of the cell spheroid incubator according to Example 1 of the present invention.
  • Figure 3 is a plan view showing a cell spheroid incubator according to Example 1 of the present invention.
  • Figure 4 is a partial cross-sectional view for showing the cap portion of the cell spheroid incubator according to Example 1 of the present invention.
  • Example 1 of the present invention a cell spheroid incubator according to Example 1 of the present invention will be described with reference to FIGS. 1 to 4.
  • the cell spheroid incubator 100 includes a body part 110, a filter part 130, and a cap part 150.
  • Body portion 110 has an opening in the upper side, it may be in the form of a tube containing the cell culture.
  • the filter unit 130 may be installed inside the body 110 to be detachably attached to the body 110.
  • the filter unit 130 may include a skeletal member 133 and a mesh 131 having a mesh shape.
  • the skeletal member 133 may be manufactured in a lattice shape, and the mesh net 131 may be installed in a lattice-shaped empty square portion of the skeletal member 133 and may be installed along an inner wall of the body part 110. (See FIG. 2).
  • Mesh-type mesh 131 may be manufactured to have a predetermined eye size as needed. Accordingly, it is possible to determine whether the cell spheroid passes by the predetermined size.
  • the cap 150 may be coupled to the opening of the body 110 to block the opening of the body 110.
  • Cap portion 150 may be coupled (or screwed) to the body portion 110 in the form of a screw, and thus may be more closely coupled.
  • the cap part 150 may include a protrusion 155 protruding in the direction toward the body part 110 (see FIG. 2).
  • the protrusion 155 may serve to fix the filter unit 130 by fitting the filter unit 130 inward.
  • the cap part 150 when the cap part 150 is coupled to the body part 110, the cap part 150 not only serves to block the opening of the body part 110, but also tightly grips the filter part 130 by the protrusion 155. Accordingly, shaking of the filter unit 130 may be prevented and the filter unit 130 may be firmly fixed.
  • At least one through hole 160 may be formed in the cap part 150 to enable gas supply to the inside of the body part 110.
  • the inflow of external gas can be controlled through the through hole 160.
  • the through hole 160 may serve as a passage for allowing air to enter and exit the body 110 when necessary in the culture process.
  • the cap part 150 includes a second cap part 153 rotatably coupled to the first cap part 151 and the first cap part 151 about the central axis O. It may include.
  • the first cap part 151 may have a plurality of through holes of different sizes.
  • the through holes 161 of the four first cap portions which increase in size as the clockwise goes, are illustrated.
  • the second cap part 153 may have a through hole formed at a position corresponding to the through hole formed in the first cap part 151.
  • the through hole 163 of the second cap portion having a corresponding size is shown at a position corresponding to the four through holes of the first cap portion 151.
  • the number of the through holes opened may be adjusted in such a way that some of the four through-holes 161 of the first cap part are opened and a part thereof is blocked.
  • the amount of gas supplied to the inside can be adjusted. For example, when culturing cells under conditions such as hypoxic conditions, the inflow of oxygen may be lowered by reducing the degree of opening and closing of the through hole 160 provided in the cap 150.
  • the through hole 160 provided in the cap part 150 may be a passage through which air may enter and exit the cell, and at this time, a filtration filter (not shown) may be installed inside the through hole. Filtration filter prevents the microorganisms contained in the air from entering the culture medium through the air passage, thereby allowing only air to enter and exit.
  • the filter unit 130 is wrapped in a mesh net 131 is coupled within the body portion 110 can prevent the cells of different sizes from being mixed between the inside and the outside of the filter unit 130. .
  • the filter unit 130 has a mesh form of a mesh
  • the cell spheroid formed inside the filter unit 130 is located inside the filter unit 130 and cannot go out, but the size is small.
  • the cells may be mixed into and out of the filter unit 130.
  • the filter unit 130 of the cell spheroid incubator 100 may be easily separated from the body unit 110. Therefore, it can be very convenient to separate only the filter unit 130 formed with the three-dimensional cell spheroid to move to other culture conditions. In addition, the separation of the cell spheroids and single cells that do not form a spheroid may be easy.
  • the cell spheroid incubator 100 includes a body part 110 containing a cell culture solution and a filter part 130 to form a three-dimensional cell spheroid inside the body part 110. And, by having a cap 150 to block the opening of the body portion 110 to control the inflow of external gas, it is possible to perform the three-dimensional cell spheroid culture more easily, such as significantly simplifying the mounting and recovery process of the cells Can provide cell spheroids in high yield, and can further lower the cost of producing cell spheroid cultures.
  • 5 is a perspective view showing a rotator of a cell spheroid culture system according to Example 2 of the present invention.
  • 6 is a perspective view showing a chamber of the cell spheroid culture system according to Example 2 of the present invention.
  • 7 is a perspective view showing a cell spheroid culture system according to Example 2 of the present invention.
  • Cell spheroid culture system according to Example 2 of the present invention comprises a cell spheroid incubator according to Example 1 described above.
  • Example 2 is different from Example 1 in that it further includes a rotator and a chamber.
  • Example 2 of the present invention a cell spheroid culture system according to Example 2 of the present invention will be described with reference to FIGS. 5 to 7.
  • the cell spheroid culture system 1000 includes the cell spheroid incubator 100, the rotator 210, and the chamber 230 according to the first embodiment.
  • the rotator 210 may include a mounting portion 211 on which the cell spheroid incubator 100 is mounted, and rotate the cell spheroid incubator 100.
  • the rotator 210 may rotate the cell spheroid incubator 100 in the R direction and adjust the rotation speed thereof.
  • the chamber 230 may accommodate the rotator 210 equipped with the cell spheroid incubator 100 in the inner space 231 of the chamber.
  • the chamber 230 may be made of acrylic or another material, and may withstand a pressure of 1 bar and maintain a predetermined pressure by adjusting to a desired pressure.
  • the chamber 230 may adjust an internal temperature, and may constantly control the cell culture conditions of 37 degrees Celsius.
  • the chamber 230 may also have waterproofing means to prevent unnecessary liquids from penetrating into the chamber 230.
  • the chamber 230 may be installed outside the chamber 230 and may include a controller 233 for adjusting the rotational speed of the rotator 210 accommodated in the chamber 230 (see FIG. 7).
  • the controller 233 is connected to the rotator 210 in a wired or wireless manner so as to transmit and receive an electrical signal with the rotator 210, and thus the chamber 230 outside the chamber 230 without opening the chamber 230. Rotational speed of the cell spheroid incubator 100 inside) can be adjusted as needed.
  • the cumbersomeness can be reduced, and the rotation speed of the cell spheroid incubator 100 is controlled without opening the chamber 230, thereby preventing the environment in which the cells are cultured inside the chamber 230. May not be received and thus the production efficiency and productivity of the cell spheroids may be significantly improved.
  • the cell spheroid culture system 1000 can perform cell spheroid culture more easily, provide a cell spheroid with high yield, and further lower the cell spheroid culture production cost. Can be.
  • FIG 8 is a view showing the appearance of the 293 cell spheroid produced through the cell spheroid culture system 1000 according to Example 2 of the present invention.
  • 9 is a view showing the appearance of various other cell spheroids produced through the cell spheroid culture system 1000 according to the second embodiment of the present invention.
  • a culture solution containing 293 cells was accommodated in the cell spheroid incubator 100, installed in the rotator 210, and rotated at 5 rpm, and placed in a chamber 230 at 37 ° C., 5%, and CO 2 to incubate the cells. .
  • the state of the spheroid produced was observed through an optical microscope.
  • the 293 cell spheroid thus produced is shown in FIG. 8.
  • FIG. 10 is a front view showing the cell spheroid culture system according to Example 3 of the present invention.
  • Fig. 11 is a left side view showing the cell spheroid culture system according to Example 3 of the present invention.
  • 12 is a right side view showing the cell spheroid culture system according to Example 3 of the present invention.
  • FIG. 13 is a conceptual diagram showing an operating mechanism of the cell spheroid culture system 3000 according to Embodiment 3 of the present invention.
  • Example 3 differs from Example 2 in that the chamber comprises a control unit and a culture unit.
  • the same (or substantial) reference numerals will be given to the same (or substantial) parts as those described above, and detailed description thereof will be omitted.
  • Example 3 of the present invention a cell spheroid culture system according to Example 3 of the present invention will be described with reference to FIGS. 10 to 13.
  • a chamber in the cell spheroid culture system 3000 according to Embodiment 3 of the present invention, includes a controller 333 and a culture unit 335.
  • the culture unit 335 is a portion in which the rotator 310 is accommodated, and may mean a portion in which cells are cultured.
  • the control unit 333 may have a feature that is installed in isolation from the culture unit 335. Accordingly, the speed control of the rotator 310 inside the culture unit 335 can be performed outside the culture unit 335. Therefore, the speed of the rotator 310 inside the culture unit 335 can be adjusted without opening and closing the inlet of the culture unit 335 to the outside. As a result, the cell culture environment inside the culture unit 335 may be smoothly performed without being disturbed.
  • the cell culture environment inside the culture unit 335 may be affected by the external environment to provide an appropriate cell culture environment. You will not be able to maintain it. In this case, the efficiency of cell culture may be reduced.
  • the rotator 310 installed in the culture unit 335 may include a motor 313 and a rotation shaft connected to the motor 313 (see FIG. 13).
  • the mounting part 311 may be installed on the rotating shaft (see FIG. 11).
  • the cell spheroid incubator 100 may be installed in the mounting portion 311.
  • the mounting portion 311 rotates, and accordingly, the spheroid incubator 100 also rotates.
  • the controller 333 may include a control connector 343 and a main controller 341.
  • the control connector 343 may be electrically connected to the motor 313 inside the rotator 310 to recognize the rotational speed of the motor 313.
  • the main controller 341 may be connected to the control connector 343 to exchange electrical signals with the control connector 343, and may control the rotational speed of the motor identified from the control connector 343.
  • the control connector 343 may recognize this and transfer this information to the main controller 341.
  • the main controller 341 may transmit a signal to the motor 313 to make the speed of the motor 313 higher, and the motor 313 receiving this signal may speed up the rotation speed of the mounting part 311. can do.
  • the main controller 341 may also be connected to the other temperature sensor 345 and the pressure sensor 347 to control the temperature and pressure inside the culture unit 335. In this case, the main controller 341 may simultaneously control the rotation speed of the rotator 310, the motor 313, the temperature inside the culture unit 335, and the pressure inside the culture unit 335. In this case, the environment inside the culture unit 335 may be a more suitable environment for cell spheroid culture.

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Abstract

The present invention relates to a cell spheroid cultivator, and a cell spheroid culture system containing the same and, more specifically, to a cell spheroid cultivator which can carry out, more conveniently, cell spheroid culture, provide cell spheroid in a high yield, and further lower cell spheroid culture production costs, and a cell spheroid culture system containing the same. According to the present invention, the cell spheroid cultivator comprises: a body part having an aperture and containing a cell culture solution; a filter part comprising a mesh-type net provided along the inner wall of the body part, and provided in the inside of the body part so as to be able to be separated from the body part; and a cap part coupled to the aperture of the body part, and having a through-hole for gas supply.

Description

세포 스페로이드 배양기 및 이를 포함하는 세포 스페로이드 배양 시스템Cell spheroid incubator and cell spheroid culture system comprising the same
본 발명은 세포 스페로이드 배양기 및 이를 포함하는 세포 스페로이드 배양 시스템에 관한 것으로서, 보다 자세하게는 세포 스페로이드 배양을 보다 간편하게 수행하고, 고수율로 세포 스페로이드를 제공할 수 있으며, 세포 스페로이드 배양 생산단가를 더욱 낮출 수 있는 세포 스페로이드 배양기 및 이를 포함하는 세포 스페로이드 배양 시스템에 관한 것이다.The present invention relates to a cell spheroid incubator and a cell spheroid culture system including the same, and more particularly, to perform cell spheroid culture more easily, to provide a cell spheroid with high yield, and to produce cell spheroid culture. The present invention relates to a cell spheroid incubator and a cell spheroid culture system including the same, which can further lower the unit cost.
세포 배양이라 함은 생물체로부터 분리한 세포를 배양하는 과정을 의미한다. 생체조직을 무균적으로 선발해서 트립신이나 프로나아제 등의 소화효소로 처리하여 단세포로 분리하여 초대배양을 하고, 또 계대중인 세포계나 세포주를 같은 효소처리로 분산시켜 얻어낸 단세포를 증식배지에 이식, 접종하여 다음의 계대배양을 하는 등 단백질 분해효소 등의 처리로 단세포를 분산해서 배양하는 방법 등이 있다.Cell culture refers to a process of culturing cells isolated from an organism. Aseptically select biological tissues, treat them with digestive enzymes such as trypsin or pronase, separate them into single cells, perform primary cultures, and transplant single cells obtained by dispersing passaged cell lines or cell lines with the same enzyme treatment to the growth medium. There is a method of dispersing and culturing single cells by treatment with proteolytic enzymes such as inoculation and subsequent passage.
일반적으로 세포 배양 방법은 2차원으로 된 평면 용기에서 단층 세포의 배양을 시행하고 있다. 이러한 2차원적인 세포 배양 및 모델링을 통해 세포 운동 및 전염과 질병의 메커니즘에 대한 주요한 통찰을 제공받을 수 있다.In general, the cell culture method is to culture monolayer cells in a two-dimensional flat container. This two-dimensional cell culture and modeling provides key insights into the mechanisms of cell motility and transmission and disease.
그러나 체내 세포 및 조직은 아주 복잡한 3차원 구조로 상호 작용하면서 성장하고 분화/ 발전해 간다. 뿐만 아니라 우리의 몸을 침입하고 전염병을 유발하는 생물 병원체들도 복잡한 3차원 조직 환경과 긴밀하게 상호 작용하게 된다. 이러한 점을 고려해볼 때, 2차원 세포 배양은 인체 내 특이적인 환경을 모사하지 못한다는 결정적인 문제점을 가지고 있다. However, cells and tissues in the body grow, differentiate, and develop as they interact with highly complex three-dimensional structures. In addition, biological pathogens that invade our bodies and cause epidemics also interact closely with complex three-dimensional tissue environments. In view of this, two-dimensional cell culture has a critical problem that does not simulate the specific environment in the human body.
따라서 최근에는 생체 내와 유사한 기능을 가지는 3차원 세포 스페로이드(cell spheroid) 배양 기술에 관한 연구가 주목 받고 있다.Therefore, in recent years, research on a three-dimensional cell spheroid culture technology having a similar function in vivo has attracted attention.
이와 관련한 3차원 세포 배양 방법으로는 현적배양법(hanging-drop method), 회전식 배양법, 원심분리법, 마이크로 몰딩법 등이 있으며, 이와 관련된 특허로는 한국등록특허 제10-1341572호 (현적배양법), 한국등록특허 제10-1412155호(회전식 배양법), 한국등록특허 제10-1362293호 (마이크로 몰딩법) 등이 있다.Three-dimensional cell culture methods in this regard include a hanging-drop method, a rotary culture method, a centrifugal separation method, a micro molding method, and related patents, such as Korean Patent Registration No. 10-1341572 (Modern Culture Method), Korea Patent No. 10-1412155 (rotary culture method), Korea Patent No. 10-1362293 (micro molding method) and the like.
다만, 기존의 현적배양법의 경우 노동 집약적이고 대량생산이 불가능하며 세포 스페로이드를 옮기는 과정에서 세포에 물리적인 손상 및 미생물의 오염을 발생시킬 수 있는 문제가 있었다. 또한 마이크로 몰딩법의 경우 제작 과정에서 반도체 공정기반의 특수한 장비들이 필수적으로 요구되는 문제가 있었다. 또한, 다른 방법들인 회전식 배양법 등도 특수한 장비가 필요하게 되어 생산단가가 높아지는 문제가 있었다.However, in the case of the conventional culture method, there is a problem that can cause physical damage and contamination of microorganisms in the process of transferring the cell spheroid, which is labor-intensive and impossible in mass production. In addition, in the case of the micro-molding method, there was a problem that special equipment based on the semiconductor process is required in the manufacturing process. In addition, other methods, such as a rotary culture method requires a special equipment, there was a problem that the production cost increases.
따라서 본 발명은 위와 같은 문제를 해결하기 위해 안출된 것으로서, 본 발명의 과제는 세포 스페로이드 배양을 보다 간편하게 수행하고, 고수율로 세포 스페로이드를 제공할 수 있으며, 세포 스페로이드 배양 생산단가를 더욱 낮출 수 있는 세포 스페로이드 배양기 및 이를 포함하는 세포 스페로이드 배양 시스템을 제공하는 것이다.Therefore, the present invention has been made to solve the above problems, the problem of the present invention is to perform the cell spheroid culture more easily, to provide a cell spheroid with a high yield, the cell spheroid culture production cost more It is to provide a cell spheroid incubator and a cell spheroid culture system comprising the same.
본 발명에 따른 세포 스페로이드 배양기는 개구부를 가지며, 세포 배양액이 담기는 몸체부, 몸체부의 내벽을 따라 설치되는 메쉬 형태의 그물망을 포함하고, 몸체부와 분리 가능하게 몸체부의 내부에 설치되는 필터부, 및 몸체부의 개구부에 결합되며, 가스 공급이 가능한 관통홀이 형성된 캡부를 포함한다.Cell spheroid incubator according to the present invention has an opening, the body portion containing the cell culture solution, a mesh-shaped net is installed along the inner wall of the body portion, the filter portion is installed inside the body portion detachably from the body portion And is coupled to the opening of the body portion, and includes a cap portion formed with a through hole capable of supplying gas.
본 발명에 따른 세포 스페로이드 배양 시스템은 개구부를 가지며 세포 배양액이 담기는 몸체부, 몸체부의 내벽을 따라 설치되는 메쉬 형태의 그물망을 포함하며 몸체부와 분리 가능하게 몸체부의 내부에 설치되는 필터부, 및 몸체부의 개구부에 결합되며 가스 공급이 가능한 관통홀이 형성된 캡부를 포함하는 세포 스페로이드 배양기; 세포 스페로이드 배양기가 장착되는 장착부를 구비하며 세포 스페로이드 배양기를 회전시키는 로테이터; 및 세포 스페로이드 배양기가 장착된 로테이터를 수용하는 챔버;를 포함한다.Cell spheroid culture system according to the present invention has an opening and the body portion containing the cell culture solution, including a mesh-shaped mesh is installed along the inner wall of the body portion and the filter portion is installed inside the body portion to be separated from the body portion, A cell spheroid incubator coupled to an opening of the body part and including a cap part formed with a through hole capable of supplying gas; A rotator having a mounting portion on which the cell spheroid incubator is mounted and rotating the cell spheroid incubator; And a chamber containing a rotator equipped with a cell spheroid incubator.
본 발명에 따른 세포 스페로이드 배양기는 세포 배양액이 담기는 몸체부, 메쉬 형태의 그물망을 포함하고, 몸체부의 내부에 설치되는 필터부, 개구부에 결합되며, 가스 공급이 가능한 관통홀이 형성된 캡부를 포함함으로써, 세포 스페로이드 배양을 보다 간편하게 수행하고, 고수율로 세포 스페로이드를 제공할 수 있으며, 세포 스페로이드 배양 생산단가를 더욱 낮출 수 있다. The cell spheroid incubator according to the present invention includes a body portion containing the cell culture solution, a mesh in the form of a mesh, a filter portion installed inside the body portion, and a cap portion coupled to the opening portion and having a through hole capable of supplying gas. By doing so, the cell spheroid culture can be more easily performed, the cell spheroid can be provided in high yield, and the cell spheroid culture production cost can be further lowered.
본 발명에 따른 세포 스페로이드 배양 시스템은 세포 스페로이드 배양기, 세포 스페로이드 배양기를 회전시키는 로테이터, 및 세포 스페로이드 배양기가 장착된 로테이터를 수용하는 챔버를 포함함으로써, 세포 스페로이드 배양을 보다 간편하게 수행하고, 고수율로 세포 스페로이드를 제공할 수 있으며, 세포 스페로이드 배양 생산단가를 더욱 낮출 수 있다.The cell spheroid culture system according to the present invention includes a chamber containing a cell spheroid incubator, a rotator for rotating the cell spheroid incubator, and a rotator equipped with the cell spheroid incubator, thereby making cell spheroid culture more convenient. In addition, it is possible to provide a cell spheroid with a high yield, and to lower the production cost of the cell spheroid culture.
도 1은 본 발명의 실시예 1에 따른 세포 스페로이드 배양기를 도시하는 사시도이다.1 is a perspective view showing a cell spheroid incubator according to Example 1 of the present invention.
도 2는 본 발명의 실시예 1에 따른 세포 스페로이드 배양기의 내부 구조를 도시하기 위한 사시도 및 부분 단면도이다.Figure 2 is a perspective view and a partial cross-sectional view for showing the internal structure of the cell spheroid incubator according to Example 1 of the present invention.
도 3은 본 발명의 실시예 1에 따른 세포 스페로이드 배양기를 도시하는 평면도이다.Figure 3 is a plan view showing a cell spheroid incubator according to Example 1 of the present invention.
도 4는 본 발명의 실시예 1에 따른 세포 스페로이드 배양기의 캡부를 도시하기 위한 부분 단면도이다.Figure 4 is a partial cross-sectional view for showing the cap portion of the cell spheroid incubator according to Example 1 of the present invention.
도 5는 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템의 로테이터를 도시하는 사시도이다.5 is a perspective view showing a rotator of a cell spheroid culture system according to Example 2 of the present invention.
도 6은 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템의 챔버를 도시하는 사시도이다.6 is a perspective view showing a chamber of the cell spheroid culture system according to Example 2 of the present invention.
도 7은 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템을 도시하는 사시도이다.7 is a perspective view showing a cell spheroid culture system according to Example 2 of the present invention.
도 8은 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템을 통하여 제작된 293 세포 스페로이드의 모습을 도시하는 도면이다.8 is a view showing the appearance of the 293 cell spheroid produced through the cell spheroid culture system according to Example 2 of the present invention.
도 9는 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템을 통하여 제작된 기타 여러 가지 세포 스페로이드의 모습을 도시하는 도면이다.9 is a view showing the appearance of various other cell spheroids produced through the cell spheroid culture system according to Example 2 of the present invention.
도 10은 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템을 도시하는 정면도이다. 10 is a front view showing the cell spheroid culture system according to Example 3 of the present invention.
도 11은 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템을 도시하는 좌측면도이다.Fig. 11 is a left side view showing the cell spheroid culture system according to Example 3 of the present invention.
도 12는 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템을 도시하는 우측면도이다.12 is a right side view showing the cell spheroid culture system according to Example 3 of the present invention.
도 13은 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템의 작동 메커니즘을 도시하는 개념도이다.Figure 13 is a conceptual diagram showing the operating mechanism of the cell spheroid culture system according to Example 3 of the present invention.
이하에서는 첨부의 도면을 참조하여 본 발명의 바람직한 실시예를 상세하게 설명한다. 그러나 본 발명이 이하의 실시예에 의해 제한되거나 한정되는 것은 아니다. Hereinafter, with reference to the accompanying drawings will be described in detail a preferred embodiment of the present invention. However, the present invention is not limited or limited by the following examples.
실시예Example 1 One
도 1은 본 발명의 실시예 1에 따른 세포 스페로이드 배양기를 도시하는 사시도이다. 도 2는 본 발명의 실시예 1에 따른 세포 스페로이드 배양기의 내부 구조를 도시하기 위한 사시도 및 부분 단면도이다. 도 3은 본 발명의 실시예 1에 따른 세포 스페로이드 배양기를 도시하는 평면도이다. 도 4는 본 발명의 실시예 1에 따른 세포 스페로이드 배양기의 캡부를 도시하기 위한 부분 단면도이다.1 is a perspective view showing a cell spheroid incubator according to Example 1 of the present invention. Figure 2 is a perspective view and a partial cross-sectional view for showing the internal structure of the cell spheroid incubator according to Example 1 of the present invention. Figure 3 is a plan view showing a cell spheroid incubator according to Example 1 of the present invention. Figure 4 is a partial cross-sectional view for showing the cap portion of the cell spheroid incubator according to Example 1 of the present invention.
이하에서는 도 1 내지 도 4를 참조하여 본 발명의 실시예 1에 따른 세포 스페로이드 배양기 대해 설명한다.Hereinafter, a cell spheroid incubator according to Example 1 of the present invention will be described with reference to FIGS. 1 to 4.
먼저 도 1 및 도 2를 참조하면, 본 발명의 실시예 1에 따른 세포 스페로이드 배양기(100)는 몸체부(110), 필터부(130), 및 캡부(150)를 포함한다.First, referring to FIGS. 1 and 2, the cell spheroid incubator 100 according to Embodiment 1 of the present invention includes a body part 110, a filter part 130, and a cap part 150.
몸체부(110)는 상측에 개구부를 가지며, 세포 배양액을 담는 튜브 형태일 수 있다. Body portion 110 has an opening in the upper side, it may be in the form of a tube containing the cell culture.
필터부(130)는 몸체부(110)에 탈부착하는 방식으로 분리 가능하게 몸체부(110)의 내부에 설치될 수 있다. 필터부(130)는 골격부재(133)와 메쉬(mesh) 형태의 그물망(131)을 포함할 수 있다. 골격부재(133)는 격자형으로 제작될 수 있으며, 메쉬 형태의 그물망(131)은 골격부재(133)의 격자형의 빈 네모칸 부분에 설치되며, 몸체부(110)의 내벽을 따라 설치될 수 있다(도 2 참조). 메쉬 형태의 그물망(131)은 필요에 따라서 소정의 눈 크기를 가지도록 제작할 수 있다. 그에 따라 소정의 크기 별로 세포 스페로이드의 통과 여부를 결정할 수 있다.The filter unit 130 may be installed inside the body 110 to be detachably attached to the body 110. The filter unit 130 may include a skeletal member 133 and a mesh 131 having a mesh shape. The skeletal member 133 may be manufactured in a lattice shape, and the mesh net 131 may be installed in a lattice-shaped empty square portion of the skeletal member 133 and may be installed along an inner wall of the body part 110. (See FIG. 2). Mesh-type mesh 131 may be manufactured to have a predetermined eye size as needed. Accordingly, it is possible to determine whether the cell spheroid passes by the predetermined size.
캡부(150)는 몸체부(110)의 개구부에 결합되어 몸체부(110)의 개구부를 막을 수 있다. 캡부(150)는 몸체부(110)와 스크류 형태로 결합(또는 나사 결합)할 수 있으며, 그에 따라 더욱 긴밀한 결합이 가능할 수 있다. 캡부(150)가 몸체부(110)와 긴밀하게 결합되면, 미생물의 침투가 방지될 수 있고, 몸체부(110)가 뒤집어지거나 회전하는 상황이 오더라도 배양액의 누수가 방지될 수 있다. The cap 150 may be coupled to the opening of the body 110 to block the opening of the body 110. Cap portion 150 may be coupled (or screwed) to the body portion 110 in the form of a screw, and thus may be more closely coupled. When the cap part 150 is tightly coupled with the body part 110, the penetration of microorganisms can be prevented, and even if the body part 110 is inverted or rotated, leakage of the culture solution can be prevented.
캡부(150)는 몸체부(110)를 향하는 방향으로 돌출하는 돌출부(155)를 포함할 수 있다(도 2 참조). 돌출부(155)는 필터부(130)를 내측으로 끼우는 방식으로 필터부(130)를 고정하는 역할을 할 수 있다. 이처럼 캡부(150)는 몸체부(110)와 결합 시, 몸체부(110)의 개구부를 막는 역할을 할 뿐만 아니라, 돌출부(155)에 의하여 필터부(130)를 타이트하게 잡아줄 수 있다. 그에 따라 필터부(130)의 흔들림이 방지되고 필터부(130)는 견고하게 고정될 수 있다.The cap part 150 may include a protrusion 155 protruding in the direction toward the body part 110 (see FIG. 2). The protrusion 155 may serve to fix the filter unit 130 by fitting the filter unit 130 inward. As such, when the cap part 150 is coupled to the body part 110, the cap part 150 not only serves to block the opening of the body part 110, but also tightly grips the filter part 130 by the protrusion 155. Accordingly, shaking of the filter unit 130 may be prevented and the filter unit 130 may be firmly fixed.
캡부(150)에는 몸체부(110)의 내부로 가스 공급이 가능하도록 관통홀(160)이 적어도 하나 이상 형성될 수 있다. 관통홀(160)을 통해 외부 가스 유입을 조절할 수 있다. 관통홀(160)은 배양 과정에서 필요 시 몸체부(110)의 내외부로 공기가 출입할 수 있도록 하는 통로의 역할을 할 수도 있다. At least one through hole 160 may be formed in the cap part 150 to enable gas supply to the inside of the body part 110. The inflow of external gas can be controlled through the through hole 160. The through hole 160 may serve as a passage for allowing air to enter and exit the body 110 when necessary in the culture process.
도 3 및 도 4를 참조하면, 구체적으로 캡부(150)는 제1 캡부(151) 및 제1 캡부(151)에 중심축(O)을 기준으로 회전 가능하게 결합하는 제2 캡부(153)를 포함할 수 있다. 3 and 4, in detail, the cap part 150 includes a second cap part 153 rotatably coupled to the first cap part 151 and the first cap part 151 about the central axis O. It may include.
제1 캡부(151)는 서로 다른 크기의 복수 개의 관통홀이 형성될 수 있다. 도 3에서는 시계방향으로 감에 따라 크기가 점점 더 커지는 4개의 제1 캡부의 관통홀(161)이 도시되고 있다. The first cap part 151 may have a plurality of through holes of different sizes. In FIG. 3, the through holes 161 of the four first cap portions, which increase in size as the clockwise goes, are illustrated.
제2 캡부(153)는 제1 캡부(151)에 형성된 관통홀에 대응하는 위치에서 관통홀이 형성될 수 있다. 도 3에서는 제1 캡부(151)의 4개의 관통홀에 대응하는 위치에서 대응하는 크기를 가지는 제2 캡부의 관통홀(163)이 도시되고 있다. The second cap part 153 may have a through hole formed at a position corresponding to the through hole formed in the first cap part 151. In FIG. 3, the through hole 163 of the second cap portion having a corresponding size is shown at a position corresponding to the four through holes of the first cap portion 151.
제2 캡부(153)를 회전시키면서 4개의 제1 캡부의 관통홀(161) 중 일부를 열리게 하고 일부를 막히게 하는 식으로 개방된 관통홀의 개수를 조절할 수 있으며, 이러한 방식으로 몸체부(110)의 내부로 공급되는 기체의 양을 조절할 수 있다. 예를 들어 저산소 조건 등과 같은 조건에서 세포를 배양할 경우, 캡부(150)에 구비된 관통홀(160)의 개폐 단계 정도를 줄이는 방식으로 산소의 유입 정도를 낮출 수 있다.While rotating the second cap portion 153, the number of the through holes opened may be adjusted in such a way that some of the four through-holes 161 of the first cap part are opened and a part thereof is blocked. The amount of gas supplied to the inside can be adjusted. For example, when culturing cells under conditions such as hypoxic conditions, the inflow of oxygen may be lowered by reducing the degree of opening and closing of the through hole 160 provided in the cap 150.
캡부(150)에 구비되는 관통홀(160)은 세포 배양 시 공기가 출입할 수 있는 통로가 될 수 있으며, 이때 관통홀 내부에 여과 필터(미도시)가 설치될 수 있다. 여과 필터는 공기 중에 포함된 미생물이 공기 통로를 통해 배양액 내부로 유입되지 않도록 하며, 그에 따라 공기만이 출입할 수 있도록 한다.The through hole 160 provided in the cap part 150 may be a passage through which air may enter and exit the cell, and at this time, a filtration filter (not shown) may be installed inside the through hole. Filtration filter prevents the microorganisms contained in the air from entering the culture medium through the air passage, thereby allowing only air to enter and exit.
한편, 메쉬 형태의 그물망(131)이 감싸고 있는 필터부(130)는 몸체부(110) 안에서 결합되어 필터부(130)의 안쪽과 바깥쪽 간에 서로 다른 크기의 세포가 혼합되는 것을 방지할 수 있다.On the other hand, the filter unit 130 is wrapped in a mesh net 131 is coupled within the body portion 110 can prevent the cells of different sizes from being mixed between the inside and the outside of the filter unit 130. .
또한 필터부(130)는 메쉬 형태의 그물망 형태를 구비하고 있기 때문에, 필터부(130) 안쪽에서 형성된 세포 스페로이드는 필터부(130) 안쪽에 위치하여 외부로 나가지 못하게 되며, 다만, 크기가 작은 세포는 필터부(130) 안팎으로 혼합될 수도 있다.In addition, since the filter unit 130 has a mesh form of a mesh, the cell spheroid formed inside the filter unit 130 is located inside the filter unit 130 and cannot go out, but the size is small. The cells may be mixed into and out of the filter unit 130.
본 발명의 실시예 1에 따른 세포 스페로이드 배양기(100)의 필터부(130)는 몸체부(110)에서 쉽게 분리될 수 있다. 따라서 3차원의 세포 스페로이드가 형성된 필터부(130)만을 분리하여 다른 배양 조건으로 이동시키기 매우 편리할 수 있다. 또한 세포 스페로이드와 스페로이드가 형성되지 못한 단일 세포의 분리가 용이할 수 있다.The filter unit 130 of the cell spheroid incubator 100 according to the first embodiment of the present invention may be easily separated from the body unit 110. Therefore, it can be very convenient to separate only the filter unit 130 formed with the three-dimensional cell spheroid to move to other culture conditions. In addition, the separation of the cell spheroids and single cells that do not form a spheroid may be easy.
이처럼 본 발명의 실시예 1에 따른 세포 스페로이드 배양기(100)는 세포 배양액이 담기는 몸체부(110), 몸체부(110)의 내부에 3차원 세포 스페로이드가 형성되도록 하는 필터부(130), 및 몸체부(110)의 개구부를 막되 외부 가스의 유입을 조절할 수 있는 캡부(150)를 구비함으로써, 세포의 장착과 회수과정을 현저히 간단하게 하는 등 3차원 세포 스페로이드 배양을 보다 간편하게 수행할 수 있고, 고수율로 세포 스페로이드를 제공할 수 있으며, 세포 스페로이드 배양 생산 단가를 더욱 낮출 수 있다.As described above, the cell spheroid incubator 100 according to the first exemplary embodiment of the present invention includes a body part 110 containing a cell culture solution and a filter part 130 to form a three-dimensional cell spheroid inside the body part 110. And, by having a cap 150 to block the opening of the body portion 110 to control the inflow of external gas, it is possible to perform the three-dimensional cell spheroid culture more easily, such as significantly simplifying the mounting and recovery process of the cells Can provide cell spheroids in high yield, and can further lower the cost of producing cell spheroid cultures.
실시예Example 2 2
도 5는 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템의 로테이터를 도시하는 사시도이다. 도 6은 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템의 챔버를 도시하는 사시도이다. 도 7은 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템을 도시하는 사시도이다.5 is a perspective view showing a rotator of a cell spheroid culture system according to Example 2 of the present invention. 6 is a perspective view showing a chamber of the cell spheroid culture system according to Example 2 of the present invention. 7 is a perspective view showing a cell spheroid culture system according to Example 2 of the present invention.
본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템은 전술한 실시예 1에 따른 세포 스페로이드 배양기를 포함하여 구성된다. 다만, 실시예 2는 로테이터 및 챔버를 더 구비한다는 점에서 실시예 1과 차이가 있다. Cell spheroid culture system according to Example 2 of the present invention comprises a cell spheroid incubator according to Example 1 described above. However, Example 2 is different from Example 1 in that it further includes a rotator and a chamber.
참고로 전술한 구성과 동일한 (또는 상당한) 부분에 대해서는 동일한 (또는 상당한) 도면 부호를 부여하고, 그에 대한 상세한 설명은 생략하기로 한다.For reference, the same (or substantial) reference numerals will be given to the same (or substantial) parts as those described above, and detailed description thereof will be omitted.
이하에서는 도 5 내지 도 7을 참조하여 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템에 대해 설명한다. Hereinafter, a cell spheroid culture system according to Example 2 of the present invention will be described with reference to FIGS. 5 to 7.
본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템(1000)은 상기 실시예 1에 따른 세포 스페로이드 배양기(100), 로테이터(210), 및 챔버(230)를 포함한다. The cell spheroid culture system 1000 according to the second embodiment of the present invention includes the cell spheroid incubator 100, the rotator 210, and the chamber 230 according to the first embodiment.
도 5를 참조하면, 로테이터(210)는 세포 스페로이드 배양기(100)가 장착되는 장착부(211)를 구비하며 세포 스페로이드 배양기(100)를 회전시킬 수 있다. 로테이터(210)는 세포 스페로이드 배양기(100)를 R방향으로 회전시킬 수 있으며 그 회전 속도를 조절할 수도 있다.Referring to FIG. 5, the rotator 210 may include a mounting portion 211 on which the cell spheroid incubator 100 is mounted, and rotate the cell spheroid incubator 100. The rotator 210 may rotate the cell spheroid incubator 100 in the R direction and adjust the rotation speed thereof.
도 6 및 도 7을 참조하면, 챔버(230)는 세포 스페로이드 배양기(100)가 장착된 로테이터(210)를 챔버의 내부 공간(231)에 수용할 수 있다. 챔버(230)는 아크릴 혹은 다른 재질로 구성될 수 있으며, 1bar 의 압력을 견디고, 원하는 압력으로 조정하여 소정의 압력을 유지할 수도 있다. 또한, 이산화탄소나, 공기의 유입커넥터를 장착하여 공기 유출입을 조절할 수도 있다.6 and 7, the chamber 230 may accommodate the rotator 210 equipped with the cell spheroid incubator 100 in the inner space 231 of the chamber. The chamber 230 may be made of acrylic or another material, and may withstand a pressure of 1 bar and maintain a predetermined pressure by adjusting to a desired pressure. In addition, it is also possible to control the air inlet and outlet by installing a carbon dioxide or air inlet connector.
그리고 챔버(230)는 내부의 온도를 조절할 수 있으며, 섭씨 37도의 세포 배양 조건을 일정하게 제어할 수도 있다. 챔버(230)는 챔버(230) 내부로 불필요한 액체가 침투되지 않도록 방수 수단도 가질 수 있다.In addition, the chamber 230 may adjust an internal temperature, and may constantly control the cell culture conditions of 37 degrees Celsius. The chamber 230 may also have waterproofing means to prevent unnecessary liquids from penetrating into the chamber 230.
한편, 챔버(230)는 챔버(230)의 외부에 설치되며, 챔버(230)의 내부에 수용된 로테이터(210)의 회전 속도를 조절하는 제어부(233)를 구비할 수 있다(도 7 참조).On the other hand, the chamber 230 may be installed outside the chamber 230 and may include a controller 233 for adjusting the rotational speed of the rotator 210 accommodated in the chamber 230 (see FIG. 7).
제어부(233)는 로테이터(210)와 전기적 신호를 주고 받을 수 있도록 로테이터(210)와 유선 또는 무선으로 연결되어 있으며, 그에 따라 챔버(230)를 개방하지 않고도 챔버(230)의 외부에서 챔버(230) 내부에 있는 세포 스페로이드 배양기(100)의 회전 속도를 필요에 따라 조절할 수 있다. The controller 233 is connected to the rotator 210 in a wired or wireless manner so as to transmit and receive an electrical signal with the rotator 210, and thus the chamber 230 outside the chamber 230 without opening the chamber 230. Rotational speed of the cell spheroid incubator 100 inside) can be adjusted as needed.
이와 같은 챔버(230)를 이용하는 경우 번거로움을 줄일 수 있을 뿐만 아니라 챔버(230)의 개방 없이 세포 스페로이드 배양기(100)의 회전 속도가 제어되므로 챔버(230) 내부에서 세포가 배양되는 환경이 방해 받지 않을 수 있으며 그에 따라 세포 스페로이드의 제작 효율 및 생산성이 현저히 개선될 수 있다. In the case of using the chamber 230, the cumbersomeness can be reduced, and the rotation speed of the cell spheroid incubator 100 is controlled without opening the chamber 230, thereby preventing the environment in which the cells are cultured inside the chamber 230. May not be received and thus the production efficiency and productivity of the cell spheroids may be significantly improved.
이처럼, 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템(1000)은 세포 스페로이드 배양을 보다 간편하게 수행하고, 고수율로 세포 스페로이드를 제공할 수 있으며, 세포 스페로이드 배양 생산 단가를 더욱 낮출 수 있다. As such, the cell spheroid culture system 1000 according to the second embodiment of the present invention can perform cell spheroid culture more easily, provide a cell spheroid with high yield, and further lower the cell spheroid culture production cost. Can be.
도 8은 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템(1000)을 통하여 제작된 293 세포 스페로이드의 모습을 도시하는 도면이다. 도 9는 본 발명의 실시예 2에 따른 세포 스페로이드 배양 시스템(1000)을 통하여 제작된 기타 여러 가지 세포 스페로이드의 모습을 도시하는 도면이다.8 is a view showing the appearance of the 293 cell spheroid produced through the cell spheroid culture system 1000 according to Example 2 of the present invention. 9 is a view showing the appearance of various other cell spheroids produced through the cell spheroid culture system 1000 according to the second embodiment of the present invention.
293 세포를 포함하는 배양액을 세포 스페로이드 배양기(100)에 수용하고, 로테이터(210)에 설치하여 5rpm 조건으로 회전시키면서, 37℃, 5%, CO2 환경의 챔버(230)에 넣고 세포를 배양하였다. 이후 24시간, 48시간, 72시간 후에 제작된 세포 스페로이드를 세포 스페로이드 배양기(100)로부터 꺼내서 스페로이드가 제작된 상태를 광학현미경을 통해 관찰하였다. 이렇게 제작된 293 세포 스페로이드의 모습이 도 8에서 도시되고 있다. A culture solution containing 293 cells was accommodated in the cell spheroid incubator 100, installed in the rotator 210, and rotated at 5 rpm, and placed in a chamber 230 at 37 ° C., 5%, and CO 2 to incubate the cells. . After 24 hours, 48 hours, 72 hours after the cell spheroid produced from the cell spheroid incubator 100, the state of the spheroid produced was observed through an optical microscope. The 293 cell spheroid thus produced is shown in FIG. 8.
293세포 스페로이드 뿐만 아니라 다른 종류의 세포들도 3차원 세포 스페로이드 제작 가능 여부를 확인하기 위해 786-o, 786-VHL, HepG2, Ck-k1, NIH-3T3, 3T3L1 세포를 배양했으며 24시간 뒤에 만들어진 세포 스페로이드가 도 9에서 도시되고 있다. In addition to 293 cell spheroids, other types of cells were cultured with 786-o, 786-VHL, HepG2, Ck-k1, NIH-3T3, and 3T3L1 cells to determine whether 3D cell spheroids could be produced. The resulting cell spheroid is shown in FIG. 9.
실시예Example 3 3
도 10은 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템을 도시하는 정면도이다. 도 11은 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템을 도시하는 좌측면도이다. 도 12는 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템을 도시하는 우측면도이다. 도 13은 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템(3000)의 작동 메커니즘을 도시하는 개념도이다.10 is a front view showing the cell spheroid culture system according to Example 3 of the present invention. Fig. 11 is a left side view showing the cell spheroid culture system according to Example 3 of the present invention. 12 is a right side view showing the cell spheroid culture system according to Example 3 of the present invention. FIG. 13 is a conceptual diagram showing an operating mechanism of the cell spheroid culture system 3000 according to Embodiment 3 of the present invention.
실시예 3은 챔버가 제어부 및 배양부를 포함한다는 점에서 실시예 2와 차이가 있다. 참고로 전술한 구성과 동일한 (또는 상당한) 부분에 대해서는 동일한 (또는 상당한) 도면 부호를 부여하고, 그에 대한 상세한 설명은 생략하기로 한다.Example 3 differs from Example 2 in that the chamber comprises a control unit and a culture unit. For reference, the same (or substantial) reference numerals will be given to the same (or substantial) parts as those described above, and detailed description thereof will be omitted.
이하에서는 도 10 내지 도 13을 참조하여 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템에 대해 설명한다. Hereinafter, a cell spheroid culture system according to Example 3 of the present invention will be described with reference to FIGS. 10 to 13.
먼저 도 10 내지 도 12를 참조하면, 본 발명의 실시예 3에 따른 세포 스페로이드 배양 시스템(3000)에서 챔버는 제어부(333) 및 배양부(335)를 포함한다.First, referring to FIGS. 10 to 12, in the cell spheroid culture system 3000 according to Embodiment 3 of the present invention, a chamber includes a controller 333 and a culture unit 335.
배양부(335)는 로테이터(310)가 수용되는 부분으로, 세포의 배양이 이루어 지는 부분을 의미할 수 있다.The culture unit 335 is a portion in which the rotator 310 is accommodated, and may mean a portion in which cells are cultured.
제어부(333)는 배양부(335)와 격리되어 설치되는 것에 특징을 가질 수 있다. 그에 따라 배양부(335) 내부의 로테이터(310)의 속도 조절을 배양부(335)의 외부에서 할 수 있다. 따라서 배양부(335)의 입구를 외부로 개폐하지 않고도 배양부(335) 내부의 로테이터(310) 속도가 조절될 수 있다. 그 결과 배양부(335) 내부의 세포 배양 환경이 방해 받지 않고 세포 배양이 원활하게 이루어질 수 있다.The control unit 333 may have a feature that is installed in isolation from the culture unit 335. Accordingly, the speed control of the rotator 310 inside the culture unit 335 can be performed outside the culture unit 335. Therefore, the speed of the rotator 310 inside the culture unit 335 can be adjusted without opening and closing the inlet of the culture unit 335 to the outside. As a result, the cell culture environment inside the culture unit 335 may be smoothly performed without being disturbed.
만일 배양부(335) 내부의 로테이터(310)의 속도를 조절하기 위해서 배양부(335)의 출입문을 열어야 한다면 배양부(335) 내부의 세포 배양 환경이 외부 환경에 영향을 받아 적절한 세포 배양 환경을 유지할 수 없게 될 것이다. 이 경우 세포 배양의 효율성은 떨어질 수 있다.If the door of the culture unit 335 needs to be opened in order to control the speed of the rotator 310 inside the culture unit 335, the cell culture environment inside the culture unit 335 may be affected by the external environment to provide an appropriate cell culture environment. You will not be able to maintain it. In this case, the efficiency of cell culture may be reduced.
그리고, 배양부(335) 내부에 설치되는 로테이터(310)는 모터(313)와 모터(313)에 연결된 회전 축을 포함할 수 있다(도 13 참조). 회전축에는 장착부(311)가 설치될 수 있다(도 11 참조). 그리고 이 장착부(311)에 세포 스페로이드 배양기(100)가 설치될 수 있다. In addition, the rotator 310 installed in the culture unit 335 may include a motor 313 and a rotation shaft connected to the motor 313 (see FIG. 13). The mounting part 311 may be installed on the rotating shaft (see FIG. 11). And the cell spheroid incubator 100 may be installed in the mounting portion 311.
회전 축이 회전하면 장착부(311)가 회전하고, 그에 따라 스페로이드 배양기(100) 역시 회전하게 된다.When the rotating shaft is rotated, the mounting portion 311 rotates, and accordingly, the spheroid incubator 100 also rotates.
도 13을 참조하면, 제어부(333)는 제어 커넥터(343) 및 메인 제어기(341)를 포함할 수 있다.Referring to FIG. 13, the controller 333 may include a control connector 343 and a main controller 341.
제어 커넥터(343)는 로테이터(310) 내부의 모터(313)와 전기적으로 연결되어 모터(313)의 회전 속도를 인식할 수 있다. 그리고, 메인 제어기(341)는 제어 커넥터(343)와 전기적 신호를 주고 받을 수 있도록 제어 커넥터(343)와 연결되며, 제어 커넥터(343)로부터 파악되는 모터의 회전 속도를 컨트롤 할 수 있다. The control connector 343 may be electrically connected to the motor 313 inside the rotator 310 to recognize the rotational speed of the motor 313. In addition, the main controller 341 may be connected to the control connector 343 to exchange electrical signals with the control connector 343, and may control the rotational speed of the motor identified from the control connector 343.
구체적으로 로테이터(310)의 모터(313)가 세포 배양을 위해 적절히 정해진 속도보다 느리게 회전할 경우 제어 커넥터(343)는 이를 인식하여 이러한 정보를 메인 제어기(341)로 전달할 수 있다. 이 경우, 메인 제어기(341)는 모터(313)의 속도를 더욱 높게 만들라는 신호를 모터(313)에 전달할 수 있고, 이러한 신호를 받은 모터(313)는 장착부(311)의 회전 속도를 더욱 빠르게 할 수 있다.In detail, when the motor 313 of the rotator 310 rotates slower than a predetermined speed for cell culture, the control connector 343 may recognize this and transfer this information to the main controller 341. In this case, the main controller 341 may transmit a signal to the motor 313 to make the speed of the motor 313 higher, and the motor 313 receiving this signal may speed up the rotation speed of the mounting part 311. can do.
메인 제어기(341)는 기타 온도 센서(345)와 압력 센서(347)와도 연결되어 배양부(335) 내부의 온도와 압력을 제어할 수 있다. 이 경우 메인 제어기(341)는 로테이터(310) 모터(313)의 회전 속도, 배양부(335) 내부의 온도, 및 배양부(335) 내부의 압력을 동시에 컨트롤할 수 있다. 이 경우 배양부(335) 내부의 환경은 세포 스페로이드 배양을 위해 더욱 적절한 환경이 될 수 있다.The main controller 341 may also be connected to the other temperature sensor 345 and the pressure sensor 347 to control the temperature and pressure inside the culture unit 335. In this case, the main controller 341 may simultaneously control the rotation speed of the rotator 310, the motor 313, the temperature inside the culture unit 335, and the pressure inside the culture unit 335. In this case, the environment inside the culture unit 335 may be a more suitable environment for cell spheroid culture.
이상에서 본 발명은 비록 한정된 실시예와 도면에 의해 설명되었으나, 본 발명은 이것에 의해 한정되지 않으며, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자에 의해 본 발명의 기술사상과 아래에 기재될 특허청구범위의 균등범위 내에서 다양한 수정 및 변형이 가능함은 물론이다.Although the present invention has been described above by means of limited embodiments and drawings, the present invention is not limited thereto, and the technical concept of the present invention and the following will be described by those skilled in the art to which the present invention pertains. Various modifications and variations are possible without departing from the scope of the appended claims.

Claims (8)

  1. 개구부를 가지며, 세포 배양액이 담기는 몸체부;A body portion having an opening and containing a cell culture solution;
    상기 몸체부의 내벽을 따라 설치되는 메쉬 형태의 그물망을 포함하고, 상기 몸체부와 분리 가능하게 상기 몸체부의 내부에 설치되는 필터부; 및A filter unit including a mesh net installed along an inner wall of the body part, the filter part being installed inside the body part so as to be separated from the body part; And
    상기 몸체부의 개구부에 결합되며, 가스 공급이 가능한 관통홀이 형성된 캡부;를 포함하는 세포 스페로이드 배양기.And a cap portion coupled to the opening of the body portion and having a through hole through which gas can be supplied.
  2. 청구항 1에 있어서,The method according to claim 1,
    상기 캡부는 서로 다른 크기의 복수 개의 관통홀이 형성된 제1 캡부, 및 상기 제1 캡부에 회전 가능하게 결합하는 제2 캡부를 포함하며,The cap part includes a first cap part formed with a plurality of through holes of different sizes, and a second cap part rotatably coupled to the first cap part.
    상기 제2 캡부는 상기 제1 캡부에 형성된 관통홀에 대응하는 위치에서 관통홀이 형성되는 것을 특징으로 하는 세포 스페로이드 배양기.The second cap portion is a cell spheroid incubator, characterized in that the through hole is formed at a position corresponding to the through hole formed in the first cap portion.
  3. 청구항 1에 있어서,The method according to claim 1,
    상기 관통홀에는 여과 필터가 설치되는 것을 특징으로 하는 세포 스페로이드 배양기.Cell spheroid incubator, characterized in that the through hole is provided with a filtration filter.
  4. 청구항 1에 있어서,The method according to claim 1,
    상기 캡부는 상기 몸체부와 결합 시 상기 필터부를 고정하는 돌출부를 포함하는 것을 특징으로 하는 세포 스페로이드 배양기.The cap portion of the cell spheroid incubator, characterized in that it comprises a protrusion for fixing the filter unit when combined with the body portion.
  5. 개구부를 가지며 세포 배양액이 담기는 몸체부, 상기 몸체부의 내벽을 따라 설치되는 메쉬 형태의 그물망을 포함하며 상기 몸체부와 분리 가능하게 상기 몸체부의 내부에 설치되는 필터부, 및 상기 몸체부의 개구부에 결합되며 가스 공급이 가능한 관통홀이 형성된 캡부를 포함하는 세포 스페로이드 배양기;A body portion having an opening and containing a cell culture solution, including a mesh-shaped net installed along an inner wall of the body portion, and separated from the body portion, a filter portion installed inside the body portion, and coupled to an opening of the body portion And a cell spheroid incubator including a cap part formed with a through hole capable of supplying gas;
    상기 세포 스페로이드 배양기가 장착되는 장착부를 구비하며 상기 세포 스페로이드 배양기를 회전시키는 로테이터; 및A rotator having a mounting portion for mounting the cell spheroid incubator and rotating the cell spheroid incubator; And
    상기 세포 스페로이드 배양기가 장착된 상기 로테이터를 수용하는 챔버;를 포함하는 세포 스페로이드 배양 시스템.And a chamber containing the rotator equipped with the cell spheroid incubator.
  6. 청구항 5에 있어서,The method according to claim 5,
    상기 챔버는 상기 챔버의 내부에 수용된 상기 로테이터의 회전 속도를 조절하는 제어부를 구비하는 것을 특징으로 하는 세포 스페로이드 배양 시스템.The chamber is a cell spheroid culture system characterized in that it comprises a control unit for controlling the rotational speed of the rotator accommodated inside the chamber.
  7. 청구항 5에 있어서,The method according to claim 5,
    상기 챔버는 상기 로테이터가 수용되는 배양부; 및The chamber includes a culture unit in which the rotator is accommodated; And
    상기 배양부와 격리되어 설치되며, 상기 배양부 내부의 로테이터의 회전 속도를 조절하는 제어부를 포함하는 것을 특징으로 하는 세포 스페로이드 배양 시스템.The cell spheroid culture system, characterized in that it is installed and isolated from the culture unit, the control unit for controlling the rotational speed of the rotator inside the culture unit.
  8. 청구항 6에 있어서,The method according to claim 6,
    상기 제어부는 상기 로테이터 내부의 모터와 전기적으로 연결되어 상기 모터의 회전 속도를 인식하는 제어 커넥터; 및The control unit includes a control connector electrically connected to the motor inside the rotator to recognize the rotational speed of the motor; And
    상기 제어 커넥터와 전기적 신호를 주고 받을 수 있도록 상기 제어 커넥터와 연결되며, 상기 제어 커넥터로부터 파악되는 상기 모터의 회전 속도를 컨트롤 하는 메인 제어기;를 포함하는 것을 특징으로 하는 세포 스페로이드 배양 시스템.And a main controller connected to the control connector to exchange electrical signals with the control connector and to control the rotational speed of the motor identified from the control connector.
PCT/KR2016/010128 2015-09-18 2016-09-09 Cell spheroid cultivator, and cell spheroid culture system containing same WO2017047986A1 (en)

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