KR20170024784A

KR20170024784A - Cell Cultivation Device Capable of Identifying Cell Cultivation State

Info

Publication number: KR20170024784A
Application number: KR1020150120251A
Authority: KR
Inventors: 황준규
Original assignee: 황준규
Priority date: 2015-08-25
Filing date: 2015-08-25
Publication date: 2017-03-08

Abstract

The present invention can easily confirm the culture state of the cells through light transmittance through the natural light passed through the cell culture container and the light transmittance of the artificial light source without confirming the state of the cell culture using the biomarker, It is possible to prevent cell contamination without being exposed to the air, to maintain the pluripotency of stem cells in stem cell culturing and to obtain a sufficient amount of stem cells necessary for stem cell treatment through promotion of stem cell proliferation for a specific purpose A cell incubator capable of confirming the cell culture state using light transmittance, and a cell culture monitoring and control system using the Internet or a cloud system.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a cell culture apparatus,

The present invention can easily confirm the culture state of a cell through light transmittance without the necessity of checking the culture state of the cell by using a biomarker, prevent the cell in culture from being exposed to the air, prevent cell contamination, A cell incubator capable of confirming the cell culture state using light transmittance capable of maintaining the pluripotency of stem cells in culturing and obtaining a sufficient amount of stem cells necessary for stem cell treatment through promoting stem cell proliferation, To a cell culture monitoring and control system using the same.

In cell culture technology, a CO2 incubator (CO2 Incubator) plays a very important role in culturing cells to be applied to various bio-industries.

However, such a conventional cell incubator such as a CO2 incubator only functions to maintain the environment necessary for culturing the cells, that is, the temperature, the atmospheric pressure, and the content of CO ₂ , It has only disadvantages in that it can not perform special functions such as promoting cell culture.

Recently, cell research using a low-power laser has shown that when light is irradiated on a cell or a living tissue, energy level of a specific element is increased and the cell or tissue unit is activated to have physiological results such as promotion of cell division, promotion of tissue regeneration, .

However, such low-power laser irradiation is a process performed outside the cell incubator, and there is a high possibility that cells are contaminated from various contaminants by exposure to air, and the objectivity of the experimental results is deteriorated due to the difference in environment with the cells to be compared. And there is an inconvenience that the biomarker must be used to identify the cell culture condition.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a biomarker capable of detecting the state of culture of a cell through natural light passing through a cell culture container and light transmittance of an artificial light source, It is possible to easily confirm the culture condition of the cell, prevent the cell in culture from being exposed to the air, prevent cell contamination, maintain the pluripotency of the stem cell in the stem cell culture, and promote the stem cell proliferation, And to provide a cell culture apparatus and a cell culture monitoring and control system using the Internet or a cloud system capable of confirming a cell culture state using light transmittance capable of obtaining a sufficient amount of stem cells necessary for therapeutic treatment.

These and other objects and advantages of the present invention will become more apparent from the following description of a preferred embodiment thereof.

The above object is achieved by a light control unit fixed to a top wall of a case having a culture space, the light control unit comprising a light source for illuminating a light source and a filter slidable therein, A light control unit including at least one guide rail which is optional and a shelf having a plurality of container accommodating units arranged at a lower portion apart from the light control unit and capable of seating a cell culture container; And a PD driver disposed at a lower portion of the cell culture container and measuring a light transmittance of the light source transmitted through the cell culture container, wherein the cell culture device is capable of confirming the cell culture state using light transmittance.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to easily confirm the culture state of a cell through light transmittance without needing to confirm the culture state of the cell by using the biomarker.

1 is a perspective view of a cell incubator according to an embodiment of the present invention.
2 is a cross-sectional view of a light control unit according to an embodiment of the present invention.
3 is a perspective view of a shelf according to an embodiment of the present invention.
4 is a cross-sectional view of a guide rail having a filter according to an embodiment of the present invention.
5 is a configuration diagram of a PD driver according to an embodiment of the present invention.
6 is a plan view of a control unit according to an embodiment of the present invention.
7 is a configuration diagram of a control unit according to an embodiment of the present invention.
8 is a configuration diagram of a web-based cell culture condition monitoring and control system according to the present invention.
FIG. 9 is a configuration diagram of a cell culture condition monitoring and control system using the cloud system according to the present invention.

The present invention relates to a method for enhancing the cell differentiation and cell proliferation effect by irradiating light of a specific wavelength band to a cell, particularly a stem cell, comprising a light control part in a cell incubator and measuring light transmittance And a system capable of remotely controlling the cell culturing apparatus.

1, which is a perspective view of a cell incubator capable of confirming a cell culture state using light transmittance according to an embodiment of the present invention, and FIG. 2, which is a sectional view of a light control unit according to an embodiment of the present invention, A cell incubator 100 according to the present invention includes a light control unit 110 fixed to a top wall of a case having a culture space and includes a light source 111 for illuminating a light source and a filter 120 slidable therein, A light control unit 110 having at least one guide rail 121, 122, 123, and 124 as a light source selection type capable of selectively transmitting a specific light source, A shelf 130 disposed at a lower portion of the cell culture container 150 and having a plurality of container receiving portions 132 capable of placing the cell culture container 150, Of the light source 112, Characterized by including the PD driver 140 is to measure the amount.

The cell incubator 100 according to an embodiment of the present invention has a structure and a shape similar to those of a commonly used incubator, and a light control unit 110 is provided on a top wall inside the cell incubator 100.

The light control unit 110 includes a light source 111 for providing a light source 112, a plurality of filters 120 for adjusting a wavelength and power and guide rails 121, 122, 123 and 124 ).

The light source 111 according to the present invention may use at least one of white light, a light emitting diode (LED), and a laser diode (LD), and the wavelength band may be selected from a visible light region in near infrared rays in the range of 400 nm to 1,100 nm , And the range of the wavelength band is preferably 1 nm or less.

In addition, the filter 120 may selectively transmit only a light source having a specific wavelength or a specific intensity by using an optical filter, and may select a specific filter 120 according to an intention of the plurality of filters 120 . Only a specific filter 120 corresponding to a plurality of cells can be selected according to the cells to be cultured among the plurality of filters 120 provided. 4, which is a cross-sectional view of the guide rail having the filter according to the embodiment of the present invention, the filter 120 is mounted on the guide rails 121, 122, 123, And it is possible to selectively apply necessary filters in a plurality of cell culture containers 150 through sliding. The pair of guide rails 121, 122, 123, and 124 may include at least one pair of two rails horizontally aligned with respect to the culture space, And 124 are installed so that the filter 120 can be detached and slid.

Here, it is preferable that the energy of the light source 112 output from the light control unit 110 and reaching the cell culture container 150 is 0.3 to 16 J / cm 2. When the light source energy is less than 0.3 J / cm 2, the effect on the stem cells is insufficient and the effects such as inhibition of stem cell differentiation and promotion of cell proliferation can not be expected. If it exceeds 16 J / cm 2, And can induce DNA deformation along with cell wall destruction.

The control of the light control unit 110 may be performed by a cell culture terminal 200 connected to a cell incubator or an Internet-based remote control terminal 300 for remote control and management or a remote control terminal 300 provided in a cloud-based low- 300).

On the other hand, when the light source 111 is used as a semiconductor, the heat generated by the light emitting diode driving circuit is transmitted to the light emitting diode, which affects the operation characteristics thereof, so that the wavelength band of the light can be shifted from the ideal wavelength band. (Not shown) for cooling the heat of the light source 112 may be further included. When the light source 111 is a semiconductor laser, both the semiconductor laser and the heat generation of the circuit must be considered. The light source temperature adjusting means removes the heat of the driving circuit so that the light emitting diode can emit light in an ideal wavelength band.

The specific filter 120 may be slidable by at least one guide rail 121, 122, 123, 124 provided on the inner wall of the through frame surrounding the light control unit 110 and positioned below the light source. The pair of guide rails 121, 122, 123, and 124 may have a pair of two rails horizontally aligned with the bottom surface of the culture space, The filter 120 is installed to be detachable and slidable. At this time, the guide rails 121, 122, 123, and 124 are formed of a plurality of pairs, and the plurality of filters 120 included in the light control unit 110 can be attached. The guide rail is preferably a light source-selectable guide rail capable of natural light exposure and selectively transmitting a specific light source.

The wavelength and power of the light source 112 are controlled by the filter 120. The adjustment by the filter 120 depends on the characteristics of the filter 120 and can be controlled by the wavelength range of the filter 120, the transmittance, the reflectivity, the refractive index, have. The light wavelength and power can be adjusted by the distance between the cell culture container 150 and the filter 120.

The cell incubator 100 according to an embodiment of the present invention may further include a cover or door 160 configured to be hermetically closed so that the shelf 130 or the cell culture container 150 can be easily opened and closed . The door 160 can be opened and closed by configuring the hinge 161, and any other openable and closable means is not limited. Further, the inside can be composed of a heat insulating box member, and a culture space is formed in the inside thereof to provide a culture environment necessary for cell culture. Here, the culture environment necessary for cell culture may include a constant temperature, humidity, gas (carbon dioxide) concentration, and the like necessary for culturing cells, especially stem cells. Such a culture environment can be controlled by the state control unit 180 connected to the MCU 170.

The shelf 130 according to an embodiment of the present invention has protrusions 131 on the right and left sides and the protrusions 131 are configured to be slidable up and down through the guide part 101 formed on the side of the cell incubator And the shelf 130 can be configured to move by vibration. By allowing the shelf 130 to move vertically and horizontally, it is possible to uniformly supply oxygen and other gases required for the cells to be cultured, increase the gas exchange rate, and cultivate high-quality cells. The up / down / left / right movement of the shelf 130 can be controlled by the MCU 170 described below.

Further, it may further include at least one of a shielding screen, a shielding film, a shielding film, and a shielding member for shielding the light emitted by the light source source 111 on the shelf 130. The light source provided by the light control unit 110 can be shut off. The shielding structure can be used to prevent the light source from being irradiated to a cell used as a cell or a control that does not need light irradiation during stem cell culture. In addition, the shielding structure may be provided in the culture space or the shelf 130 so as to be able to enter and exit horizontally or vertically.

The shelf 130 may have a plurality of container receiving portions 132 disposed below the light control portion 110 and capable of placing the cell culture container 150 thereon. A plurality of such shelves can be provided.

Here, the cell culture container 150 can be used without limitations, such as a test tube, a flask, a culture dish, or any other container capable of culturing cells. The stem cells which are accommodated and cultured in the container can be all kinds of human and animal stem cells such as neural epithelial stem cells, mesenchymal stem cells, and hematopoietic stem cells.

In addition, the cell incubator 100 may be provided with a gas control means (not shown) capable of appropriately controlling the concentration of air in the cell culture medium in the culture of stem cells, and may be provided with an environment suitable for stem cell culture . In particular, the gas regulating means may be circulated to maintain an appropriate O2 / CO2 ratio in the culture of the stem cells.

In addition, the cell incubator 100 may be equipped with temperature control means (not shown) for controlling the internal temperature of the cell incubator 100. The temperature control means may be a combination of a heating means and a cooling means, May be configured to indirectly control the temperature of the cell culture apparatus 100 by heating or cooling the inner wall of the cell culture apparatus 100 or heating or cooling the gas introduced through the gas regulation means to enter the cell culture apparatus 100 .

In addition, the cell incubator 100 is provided with a temperature sensor and a gas sensor, which can confirm whether the gas adjusting means and the temperature adjusting means suitably adjust the cell culture environment, (170). It is of course also possible to display the information on the cell culture terminal 200, the remote control terminal 300 and the expert terminal 400 of the system described below.

5, which is a PD driver 140 according to an embodiment of the present invention, the PD driver 140 includes an array type photodiode (PD) 140 for receiving an optical signal irradiated from a light source 111 and converting the optical signal into an electrical signal And a proportional amplifier 143, a switching multiplexer 144 and an ADC (Analog Digital Converter) 145, which are the same as the general-purpose PD driver circuit. Is omitted. The decoder may further include a decoder that is not shown in the figure, but decodes it. The PD driver 140 is connected to the MCU 170 responsible for the control function.

6 is a plan view of a control unit according to an embodiment of the present invention. 6, the control unit 150 is provided outside the cell incubator 100 to select the overall state of the cell incubator 100 and the functions of the components constituting the cell incubator 100 And control the operation of the control unit. At this time, the control unit 150 may include a display unit for displaying the cell culture environment information and an input key or a touch screen for selecting, inputting, and controlling the cell culture environment. Also, as described later, the control unit 150 may be controlled by the cell culture terminal 200 or the remote control terminal 300.

Here, the cell culture environment may include a light source and filter selection, optical wavelength and intensity selection, light irradiation time, and the like in addition to displaying sensor data such as temperature, gas concentration, and humidity of the culture space. have.

At this time, control of the cell culture environment, light source, light wavelength and intensity by the user can be controlled by the input key or the touch screen of the control unit 150, and can be confirmed by the display unit. At this time, the display unit may display the light irradiation time set by the user, the remaining time of the light irradiation operation, the light source output value, the light output characteristic (setting of a continuous wave or a pulse frequency), a light wavelength and a filter.

The operation by the input key or the touch screen may be provided for setting each mode and time and inputting or setting using the up, down, left, and right direction keys.

The MCU 170 is a component for controlling all functions of a cell incubator that can confirm the cell culture state using the light transmittance according to the present invention. Hereinafter, the present invention will be described in detail with reference to FIG. 6, which is a block diagram of a control unit according to an embodiment of the present invention. It goes without saying that the cell culture terminal 200 and the remote control terminal 300 of the system described below can also control the function of the cell incubator in conjunction with the MCU 170.

First, the optical signal received through the array type photodiode 141 is processed and transmitted to the control unit 170 for displaying the received light amount.

The filter 120 can be controlled to slide inside the guide rail 121 and the shelf 130 on which the cell culture container 150 is placed can be controlled to move vertically and horizontally.

In addition, the intensity of the light source and the exposure time are controlled until the reception amount of the initial optical signal falls below a predetermined amount, and the state of the light source (wavelength, intensity, exposure time and distance between the incubator and the light source) And controls the light control unit 110 to display and transmit the information. A certain amount of the optical signal reception amount may vary depending on the cells to be cultured and the light transmittance varying according to the amount of light reception is compared to determine the light transmittance proportional to a certain amount of light reception amount compared with the initial light transmittance, Can be amplified and cultured. In addition, a specific wavelength can be selected according to the state or differentiation degree of the cultured cell, and the degree of cell culture can be controlled by finely analyzing the light transmission and wavelength.

Also, the function of the state control unit 180 for controlling temperature, humidity and air (carbon dioxide, etc.) is controlled so that the state of the incubator in which the light source is irradiated is similar to the state of the human body, and the distance from the light source Control function.

And a function of determining whether the culture liquid is in an optimized state using the measured permeability through the PD driver 140.

According to the present invention, since the permeability of the light passing through the cell culture container 150 through the filter 120 after being irradiated by the light source 112 will be lowered if the cells are proliferated and expanded, the existing biomarkers It is not necessary to check the state of the cultured cells.

As described above, the stem cells cultured from the cell incubator capable of confirming the cell culture state using the light transmittance according to the present invention and the cells differentiated therefrom may have a wide range of applicability in the medical and non-medical fields, It can be administered to an animal or human patient to cure the disease condition.

Next, a cell culture condition monitoring and control system according to a third aspect of the present invention will be described in detail with reference to FIG. 8, which is a block diagram thereof.

The system according to the present invention mainly comprises a cell incubator 100, a cell culture terminal 200, a remote control terminal 300, an expert terminal 400, a central server 500 and a database 600, 100, the remote control terminal 300, the expert terminal 400, and the central server 500 via the Internet.

The cell incubator 100 may be according to the present invention described above but is not limited thereto. The cell incubator may be a serial port for serial communication as a wired / wireless connection means for connection with the cell culture terminal 200, an Ethernet port based on TCP IP, an optical communication port which is not affected by lightning or electromagnetic waves, And a wireless communication module that enables RF communication using a wireless signal such as WIFI. Here, by having a TCP IP-based Ethernet port and an optical communication port capable of connecting it with an optical communication cable, it can be prevented from being affected by EMC (EMI / EMP) and EMP (electromagnetic pulse) caused by lightning or strong electromagnetic waves.

The cell culture terminal 200 provides data on the cell culture condition or condition of the cell culture apparatus 100 to the central server 500 through the Internet in real time and transmits the cell culture state or condition data to the remote control terminal 300, And controls the cell incubator 100 based on control information of the cell incubator 100 provided from the Internet 500 through the Internet. The cell culture terminal 200 may include an RS232 driver for connection to the cell incubator 100, a controller, and an interface 102 for network connection.

The interface 102 is an Internet interface when the cell culture terminal 200 is connected to the remote control terminal 300, the expert terminal 400 or the central server 500 via the Internet and an Ethernet interface when the cell culture terminal 200 is connected through the Ethernet . This system can also be implemented in a clouding environment.

In addition, the cell culture terminal 200 may include various memories such as an ordinary PC, and an SD memory card for realizing an expert system based on an expert knowledge base based on RDR (Ripple Down Rule) ) Port to the control unit.

Here, the cell culture terminal 200 and the cell incubator 100 may include an optical communication module using a glass optical fiber or a plastic optical fiber, a wireless communication module for RF communication such as ZigBee, Bluetooth or WIFI, Communication method can be adopted. The cell incubator 100 can be designed and manufactured as an embedded server based on Net + 50 (NetSilicon, Inc.), a network microprocessor in a SOC (System on Chip) format.

The remote control terminal 300 provides information to a terminal such as a person operating a cell incubator at a remote site to monitor and control the cell incubator 100 according to the cell culture condition or condition through the Internet at a remote site. That is, based on the real-time information of the cell incubator 100 provided in the central server 500, the cell culture incubator is monitored, and the specific wavelength and natural light are combined to check the cell culture condition or condition, 200 to control the cell incubator 100.

In addition, the expert terminal 400 provides information for technical control of the cell incubator 100 via the Internet at a remote site as a terminal for experts in cell culture.

The central server 500 integrates the cell culture terminal 200, the remote control terminal 300 and the expert terminal 400 via the Internet and can monitor and control the cell culture apparatus 100 in real time. In addition, the specific wavelength to be irradiated and the natural light are merged to monitor the cell culture condition in the cell incubator, and to store the culturing conditions for the specific cell, thereby enabling customized control of the specific cell. The central server may enable connection to the IP address of the cell incubator based on the information in the database.

The database 600 includes an operation information database in which various data necessary for operation of the cell culture terminal 200, the remote control terminal 300, the expert terminal 400 and the central server 500 are stored, A specific cell information DB storing the IP address of the cell incubator, and a culture information database storing cell culture information such as specific cell basic information and specific cell culture conditions.

The system according to the present invention configured as described above is for realizing cell culture smoothly out of time and space constraints by organically connecting specific cells, cell cultures, cultivating operators and experts, To be monitored and controlled from a remote place.

The remote monitoring terminal 300, the expert terminal 400 and the central server 500 can perform monitoring and control of the cell incubator 100 at a remote location through the cell culture terminal 200, The remote control terminal 300, and the expert terminal 400 through the web. The remote control terminal 300 and the expert terminal 400 can share cultured data of specific cells and information on the cell incubator 100 according to the integration function via the web provided by the central server 500, It is possible to monitor and control the culturing condition for specific cells in the cell incubator 100 through the cell culture terminal 200. [

9, the cell culture monitoring and control system according to the present invention may be a cloud system in which the Internet includes a control module, and in this case, At least one of the cell culture terminal, the database, the central server, the remote control terminal and the expert terminal can be controlled by the control module of the cloud system.

Therefore, the cell culture terminal, the remote control terminal, and the expert terminal need not have their own control unit, and can input and receive commands to the control module of the cloud system (e.g., a keyboard, a portable terminal (E.g., a touch screen or the like) and an output means (e.g., a display unit). Therefore, the present invention enables cloud-based remote management and control, and has the advantage of minimizing power consumption.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope of the invention as set forth in the claims.

100: cell incubator 111: light source
110: light control unit 121, 122, 123, 124: guide rail
120: filter 130: shelf
140: PD driver 101: Guide part
150: cell culture container 160: door
170: MCU 200: cell culture terminal
300: Remote control terminal 400: Expert terminal
500: central server 600: database

Claims

A light control unit (110) fixed to a top wall of a case having a culture space, comprising: a light source (111) for illuminating a light source; a filter (120) slidable therein; A light control unit 110 having at least one guide rail 121, 122, 123, and 124 as a light source selection type that can transmit light,
A shelf 130 disposed below the light control unit 110 and having a plurality of container receiving portions 132 for placing the cell culture container 150 thereon,
And a PD driver (140) disposed below the vessel (130) to measure a light transmission amount of the light source (112) transmitted through the cell culture container, wherein the PD driver A cell incubator to check the condition.