KR100870982B1 - A multilayer Bio-reactor - Google Patents

Abstract

In the multiple biological reaction apparatus according to the present invention, a plurality of plate members having a predetermined opening shape are stacked to form a microchannel through which a material required for cell culture passes and a space region in which cells are cultured, and to control a cell culture environment. It is characterized in that the electrical circuit wiring is embedded between the laminated plate member. According to this structure, since the entire structure is formed by stacking a ceramic panel having a thin thickness and a small size, the size of the bioreactor can be reduced in size and weight, and can be used for single use. As it provides a micro channel and a culture space, various applications are possible in designing a flow path and a space layout, and in particular, the flow path can be made three-dimensionally to make the bioreactor smaller.

Bioreactors, cell incubators, culture chambers, ceramic panels, cell therapies.

Description

Multilayer Bio-reactor

1 is a view showing the overall configuration of a bioreactor according to an embodiment of the present invention,

2 to 4 is a view showing a ceramic panel constituting a bioreactor according to an embodiment of the present invention,

5 is a view showing a configuration in which the ceramic panel is coupled according to an embodiment of the present invention,

Figure 6 is a view for explaining the cell culture portion of the bioreactor according to an embodiment of the present invention,

7 is a view showing the culture chamber shape of the bioreactor according to the embodiment of the present invention,

8 and 9 is a view showing the principle that the gas is supplied for the culture in accordance with an embodiment of the present invention,

10 is a cross-sectional view of a bioreactor according to another embodiment of the present invention;

11 is a view showing a cell culture system equipped with a bioreactor according to an embodiment of the present invention.

<Description of Major Symbols in Drawing>

100. Ceramic panel 103. Gas flow path

104. Medium flow path 105. Culture chamber

106. Gas supply unit 107. Gas permeable membrane

108. Sensor unit 109. Environment control unit

110. Wireless communication section 111. Circuit wiring

201.Concave Groove 202. Through Hole

203. Openings 601. Buffer wells

602. Outlet outlet 603. Outlet outlet

605. Heat generator 612. Contact point

610. Output badge 611. Output badge

700. Culture system 701. Bioreactor

702. Display unit 903. Porous plate member

The present invention relates to a bioreactor for cell culture. More specifically, the present invention relates to a multilayer bioreactor in which a plurality of plate members having a predetermined opening pattern are stacked up and down to form a predetermined structure, and microchannels and spaces required for cell culture are formed inside the structure.

Bioreactot is a system that uses in vitro the chemical reactions that occur in the body of the organism is also called a bioreactor. Living organisms use various enzymes in the body to break down substances or synthesize substances necessary for life support. The decomposition and synthesis reactions are all carried out at room temperature and atmospheric pressure, efficiently producing what is suitable for our body. By realizing this process in the living body to an external device, it is possible to make an ideal production that achieves resource saving and energy saving.

On the other hand, cell therapy refers to a technology for treating cancer or regenerating tissues with deteriorated function using cells having a therapeutic effect instead of conventional drugs. Representative cell therapies are techniques such as treating cancer with immune cells such as white blood cells or regenerating cartilage or heart muscle using stem cells. In particular, the brain, spinal cord, heart muscle, pancreas, etc., once damaged can not be regenerated, the cells that can be treated by stem cell culture outside the body, and then injected into the affected area is very effective in regenerating damaged organs.

For these cell therapies it is important to cultivate cells with therapeutic effects outside the body. In particular, the cultured cells should be injected again into the human body, so the tissue compatibility should be sufficiently considered. The best method is to culture the autologous or allogeneic cells in small quantities in vitro in an environment very similar to the body environment of the patient to be administered and then administer them to the patient. In addition, it is preferable to use a new incubator set to an optimized environment for each cell culture without reusing the incubator used for cell culture.

However, the conventional cell culture is mainly intended to cultivate a large amount of cells is not suitable for culturing a small amount of cells to be used as a cell therapy. In addition, since cells to be used as cell therapeutics have to have very delicate environmental control for culture, it is not suitable to culture them using general cell culture techniques.

Accordingly, there is a need for bioreactors for light and miniaturized cell cultures that can cultivate cells suitable for cell therapy by proliferating and differentiating small amounts of cells in a similar environment to the living environment, and in particular, can be used and discarded once.

The present invention has been made in view of the above, a plurality of plate-like members having a predetermined opening shape is stacked up and down to form a constant structure, the micro-channel and space required for cell culture inside the structure is formed Therefore, the object of the present invention is to provide a bioreactor for cell culture, which provides an optimized environment for cell therapy and is small in volume and disposable.

In the multi-biological reaction apparatus according to the present invention for achieving the above object, a plurality of plate-like members having an opening shape are stacked to form a microchannel through which a material required for cell culture passes and a space region in which cells are cultured, and a temperature, PH The electric circuit wiring 111 for controlling the cell culture environment including the dissolved oxygen amount (DO), the culture medium and the composition of the gas to be optimized for cell culture is characterized in that it is embedded between the laminated plate members.
The opening shape of the plate member is preferably formed by including one or two or more through-holes 202, vertical openings 203, or recessed grooves 201 dug inwardly.
The plate member is preferably a ceramic panel.
The bioreaction apparatus includes a culture chamber 105 in which a culture medium and a gas required for cell culture are supplied to proliferate cells, and a medium inlet 602 into which the culture solution is introduced and a medium outlet 603 through which the culture solution flows out; A medium input passage 610 connected to the medium inlet 602 to supply the culture solution; A medium output passage 611 connected to the medium outlet 603 to outflow the culture solution; A gas permeable membrane 107 formed under the culture chamber 105; A gas flow passage 103 through which gas necessary for cell culture passes; And a gas supply part 106 connected to the gas flow path 103 and formed under the gas permeable membrane to supply gas to the culture chamber 105.
A sensor unit 108 for detecting a cell culture environment including temperature, PH, dissolved oxygen amount (DO), a culture medium and a composition of a gas and outputting a predetermined sensor signal; Environmental control unit 109 for receiving a sensor signal or an external control signal of the sensor unit 108 to adjust the culture culture including the temperature, PH, dissolved oxygen (DO), the culture medium and the composition of the gas to optimize the cell culture ; And, connected to the sensor unit 108 and the environmental control unit 109, transmits the sensor signal of the sensor unit 108 to the outside, or receives an external control signal applied to the environmental control unit 109 The wireless communication unit 110 is further included, and the circuit wiring 111 connecting the respective units is preferably arranged between the laminated plate members.
The culture chamber 105 has the medium inlet 602 formed at a position lower than the medium outlet 603, and the width of the medium inlet 602 toward the medium inlet 602 and the medium outlet 603 is narrowed toward the medium inlet 602 and the medium outlet 603. The culture is preferably formed to have a uniform laminar flow.
A buffer well 601 formed at a front end of the culture chamber 105 to temporarily store the culture solution; The heat generating unit 605 is installed inside the buffer well 601 and heats the culture solution temporarily stored in the buffer well 601;
The culture chamber 105 is divided into an upper culture chamber and a lower culture chamber based on a predetermined porous plate member, and the feeder cells and the target cells are separated and cultured based on the porous plate member.
The porous plate-like member is preferably microfiber.

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Hereinafter, with reference to the accompanying drawings will be described in detail a multi-bioreactor according to a preferred embodiment of the present invention.

1 is a cross-sectional view of a bioreactor according to an embodiment of the present invention.

Referring to FIG. 1, a bioreactor according to an exemplary embodiment of the present invention may include a culture chamber 105, a medium passage 104, a gas permeable membrane 107, a gas passage 103, a gas supply unit 106, and a sensor unit. 108, an environmental control unit 109, a wireless communication unit 110, and a circuit wiring 111.

In particular, the bioreactor according to the embodiment of the present invention is configured by stacking a plurality of ceramic panels 100 having a predetermined opening shape up and down. FIGS. 2 to 4 illustrate the ceramic panel 100.

As shown in FIGS. 2 to 4, each ceramic panel 100 has an opening shape such as a through hole 202 drilled up and down, an opening 203 having a predetermined portion opened, or a concave groove 201 dug inward. Has The opening shape formed in each ceramic panel 100 can be variously changed according to the purpose of cell culture.

As shown in FIG. 5, when a plurality of ceramic panels 100 are stacked and coupled together, opening shapes formed differently are three-dimensionally connected to each other to form a predetermined micro flow path 101 (see FIG. 1) and a space region 102 (FIG. 1). Reference). That is, the through holes 202 and the concave grooves 201 formed at different positions and sizes form a micro flow path 101, and the openings 203 form a constant spatial region 102 (FIG. 6). Reference). At this time, each ceramic panel 100 is a plate-shaped module (for example, a silicon substrate) of a ceramic material is used, the thickness is preferably 1.5mm or less.

In FIG. 1, the micro flow path 101 and the space region 102 are formed by the opening shape on the ceramic panel 100 as described above. At this time, the culture chamber 105 supplies the nutrient substance to the cells to be cultured by passing the culture passage 104 through the culture chamber 105 to where the cells are cultured, that is, proliferated and / or differentiated. In addition, the gas required for cell culture passes along the gas passage 103 and is supplied to the culture chamber 105 through the gas permeable membrane 107 in the gas supply unit 106.

The bioreaction device according to the embodiment of the present invention may be useful for culturing cells used as cell therapeutics, and the culture environment must be precisely controlled according to the type of cells to be cultured.

In order to adjust the culture environment, the sensor unit 108 detects the cell culture environment (eg, temperature, pH, DO, etc.) in real time to generate a predetermined sensor signal. The sensor unit 108 is preferably a semiconductor sensor (eg, ISFET) capable of sensing temperature, pH, DO, and the like.

The environmental control unit 109 performs a function of adjusting the culture environment and a control IC may be used. The environmental control unit 109 can adjust the culture environment based on a predetermined value. For example, if the culture temperature is set to 37 ° C in advance, the sensor unit 108 measures the temperature and transmits the measured value to the environmental control unit 109. If the measured temperature is 40 ° C, the environmental control unit 109 lowers the temperature by applying a predetermined signal to the temperature control device 112.

At this time, the environment controlled by the environmental control unit 109 may be variously set, such as the temperature, pH, the dosage amount or composition of the culture and gas.

In addition, the environmental control unit 109 can also adjust the culture environment in accordance with an external control signal. An external manager terminal (not shown) is wirelessly connected to the bioreactor according to the embodiment of the present invention through the wireless communication unit 110. Therefore, the wireless communication unit 110 transmits the culture environment information sensed by the sensor unit 108 to the manager terminal, and performs a function of applying the control signal of the manager terminal to the environment control unit 109. For example, in order to change the culture temperature according to the purpose of culture, it is possible to control the culture environment by operating an external terminal. To this end, the wireless communication unit 110 may be an RF chip, an infrared communication module or a wireless LAN.

In addition, the bioreactor according to the embodiment of the present invention is a three-dimensional micro-channel 101 through which the culture medium and gas supplied for cell culture is arranged in three dimensions to maximize the space efficiency. Here, the microchannel 101 is three-dimensionally arranged, for example, in FIG. 1, the media channel 104 and the gas channel 103 are formed parallel to each other on the YZ plane. It means that the flow path 103 can be formed parallel to each other on the XZ plane.

Furthermore, the various circuit wirings 111 for controlling the culture environment are disposed between the ceramic panels 100 stacked so that the size of the entire bioreactor device is reduced in size and weight.

Referring to Figure 6, it will be described the principle that the cells are cultured in the bioreactor according to an embodiment of the present invention.

In FIG. 6, the culture chamber 105 is a constant space that receives the culture solution and gas required for cell culture and proliferates the cells. The composition of the culture medium and the gas required for the cell culture may be variously set and supplied according to the type of cells to be cultured, and may include growth factors for promoting proliferation and differentiation of cells. .

In addition, the culture chamber 105 has a medium inlet 602 through which the culture solution flows and a medium outlet 603 through which the culture solution flows out, respectively, at the front and rear ends thereof. That is, the culture solution supplied through the medium input passage 610 is temporarily stored in the buffer well 601 formed at the front end of the culture chamber 105 and then introduced into the culture chamber 105 through the medium inlet 602. After passing through the culture chamber 105 to the outside through the medium output passage 611 through the medium outlet 603. Thus, while the culture fluid passes through the culture chamber 105, the cells 604 existing in the culture chamber 105 are grown by obtaining nutrients from the culture solution.

At this time, a predetermined heat generating unit 605 (for example, a heater such as a heating wire) is installed inside the buffer well 601 to supply a culture medium by heating it, thereby keeping the temperature of the culture solution flowing into the culture chamber 105 constant. It is good to adjust. Also, although not shown in detail in FIG. 6, the heat generator 605 may be connected to the environment control unit 109 (see FIG. 1) or connected to the outside by a predetermined electrical contact point 612.

In addition, as shown in Figure 7, the overall shape of the culture chamber 105 is gradually narrower from the central portion having a constant width toward both ends, the medium inlet 602 is formed at a position lower than the medium outlet 603 (That is, the medium inlet 602 is formed at the lower side of the medium outlet 603 based on the Z axis) so that the culture medium has a uniform laminar flow in the culture chamber 105 so that the culture medium is cultured. It is desirable to prevent excessive entry into 105) and cell loss.

6 again, a mixed gas of N ₂ , 0 ₂ , NH ₃ , CO _2, etc. required for cell culture in a predetermined ratio passes through the gas flow passage 103 and supplies the gas to the culture chamber 105. To this end, a gas permeable membrane 107 is formed in the lower portion of the culture chamber 105, and a gas supply 106 is formed in the lower portion of the gas permeable membrane 107. As the gas permeable membrane 107, a porous membrane or a microfiber may be used.

8 and 9 will be described in more detail the principle that the cells are supplied with the required gas using the bioreactor according to an embodiment of the present invention.

In general cell culture, there is a problem that the gas 802 is not sufficiently supplied to the cells 803 due to the culture solution 801 as shown in FIG. 8.

However, in the bioreactor according to the embodiment of the present invention, as shown in Figure 9, the gas flow path 103 is formed in the lower portion of the culture chamber 105, the gas is configured to pass through the lower end of the culture chamber 105 It is. In addition, since the gas supply unit 106 has a larger cross-sectional area than the gas passage 103, the gas passing through the gas supply unit 106 has a reduced flow rate. The gas in the gas supply 106 then rises upwards due to its nature, and the elevated air is supplied directly to the cells 604 present in the culture chamber 105 through the gas permeable membrane 107.

Meanwhile, a bioreactor according to another embodiment of the present invention will be described with reference to FIG. 10.

Referring to FIG. 10, the bioreactor according to the embodiment of the present invention includes two culture chambers 901 and 902. In addition, the porous plate-like member 903 (for example, porous membrane or microfiber, etc.) separates the two culture chambers 901 and 902. That is, the portions described in FIG. 9 centering on the porous plate member 903 are formed in the upper layer portion 904 and the lower layer portion 905 in a mirror image of each other.

Since the lower layer part 905 is the same as that described in FIG. 9, a description thereof will be omitted. As described above, the upper layer portion 904 is stacked upside down the porous plate-like member 903 by inverting the configuration of the lower layer portion 905. The culture chamber 901 of the upper layer 904 and the culture chamber 901 of the lower layer 905 are spatially separated by the porous plate-like member 903, but the culture solution is allowed to pass through the porous plate-like member 903. . At this time, the feeder cells 906 are cultured in the culture chamber 901 of the upper plate 904, that is, the upper portion of the porous plate-like member 903, and that is, the lower layer 905 of the lower layer 905. In the culture chamber 902, an object cell 907 to be cultured is cultured. Using this, it is possible to isolate and culture the feeder cells 906 and the target cells 907. The feeder cell 906 refers to a cell that provides nutrients.

11 shows a cell culture system equipped with a bioreactor according to an embodiment of the present invention.

In FIG. 11, the bioreactor 701 according to the embodiment of the present invention is detachably coupled to the culture system 700 and may be used disposable or reusable. The information sensed by the sensor unit 108 is displayed on the display unit 702, and it is possible to adjust the culture solution and the gas supplied to the bioreactor 701 based on the image displayed on the display unit 702.

Although preferred embodiments of the present invention have been described above, the present invention is not limited to the above-described specific embodiments. That is, those skilled in the art to which the present invention pertains can make many changes and modifications to the present invention without departing from the spirit and scope of the appended claims, and all such appropriate changes and modifications are possible. Equivalents should be considered to be within the scope of the present invention.

As described above, according to the multiple bioreactor according to the present invention,

First, since the entire structure is formed by stacking a ceramic panel having a thin thickness and a small size, the size of the bioreactor can be reduced in size and weight, so that it can be used for single use.

Second, because the predetermined openings formed in the ceramic panel are connected to each other to provide a micro flow path and a culture space, various applications are possible in designing the flow path and the space layout, and in particular, the flow paths are three-dimensionally arranged to make the bioreactor smaller. Can be.

Third, since the circuit wiring is disposed between the ceramic panels, the connecting line connecting various devices for controlling the culture environment is simplified.

Fourth, it is very advantageous for cell culture because the gas required for the culture is supplied directly without being disturbed by the flow of the culture solution.

Fifth, it is possible to separate only the target cells spatially while culturing the feeder cells and the target cells by separating the culture space.

Sixth, since the part for controlling the culture environment is packaged in the bioreactor and can be controlled wirelessly, there is an effect that can provide an environment optimized for culturing cells to be used for cell therapy.

Claims (9)

A plurality of plate-shaped members having an opening shape are stacked to form a microchannel through which a material necessary for cell culture passes and a space region in which the cells are cultured, and include a cell including temperature, PH, dissolved oxygen (DO), a culture solution and a gas composition. An electrical circuit wiring (111) for controlling the culture environment to be optimized for cell culture is characterized in that the multi-biological reaction device is embedded between the laminated plate member. The method of claim 1, The opening shape of the plate-shaped member is formed by including one or two or more through holes 202, vertical openings 203, a predetermined portion of the opening 203 or indented grooves 201 dug inwardly. Bioreactor. The method of claim 1, The plate-like member is a multi-bioreactor, characterized in that the ceramic panel. The method of claim 1, The biological reaction device, A culture chamber 105 in which a culture medium and a gas necessary for cell culture are supplied to proliferate a cell, and a medium inlet 602 through which the culture solution is introduced and a medium outlet 603 through which the culture solution flows out are formed; A medium input passage 610 connected to the medium inlet 602 to supply the culture solution; A medium output passage 611 connected to the medium outlet 603 to outflow the culture solution; A gas permeable membrane 107 formed under the culture chamber 105; A gas flow passage 103 through which gas necessary for cell culture passes; And, And a gas supply unit (106) connected to the gas channel (103) and formed under the gas permeable membrane to supply gas to the culture chamber (105). The method of claim 4, wherein A sensor unit 108 for detecting a cell culture environment including temperature, PH, dissolved oxygen amount (DO), a culture medium and a composition of a gas and outputting a predetermined sensor signal; Environmental control unit 109 for receiving a sensor signal or an external control signal of the sensor unit 108 to adjust the culture culture including the temperature, PH, dissolved oxygen (DO), the culture medium and the composition of the gas to optimize the cell culture ; And, It is connected to the sensor unit 108 and the environmental control unit 109, the wireless to transmit the sensor signal of the sensor unit 108 to the outside, or to receive an external control signal and apply to the environmental control unit 109 Further comprising a communication unit 110, Multiple bioreactor, characterized in that the circuit wiring (111) for connecting the respective parts are arranged between the laminated plate-like member. The method of claim 4, wherein The culture chamber 105 has the medium inlet 602 formed at a lower position than the medium outlet 603, and the width of the medium inlet 602 is narrowed toward the medium inlet 602 and the medium outlet 603 at a central portion thereof. Multiple biological reaction apparatus, characterized in that the culture medium is formed to pass through with a uniform laminar flow. The method of claim 4, wherein A buffer well 601 formed at a front end of the culture chamber 105 to temporarily store the culture solution; And a heat generator (605) installed inside the buffer well (601) to heat the culture solution temporarily stored in the buffer well (601). The method of claim 4, wherein The culture chamber 105 is divided into an upper culture chamber and a lower culture chamber with a predetermined porous plate member as the center, and a multi-organism characterized in that nutrient cells and target cells are separated and cultured, respectively, based on the porous plate member. Reaction device. The method of claim 8, The porous plate-like member is a multiple biological reaction device, characterized in that the microfiber.

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