KR102119479B1 - 3d cell culture biosensor and manufacturing method thereof - Google Patents

Abstract

A temperature sensor substrate including a temperature sensor, a plurality of impedance sensor chips and the temperature sensors and the plurality of impedance sensor chips disposed vertically with the temperature sensor substrate in a direction facing each other with the temperature sensors interposed therebetween. A 3D cell culture biosensor comprising a well for cell culture arranged to include, and measuring the impedance and temperature change of cells cultured in the cell culture well using the temperature sensor and the impedance sensor chip. Is disclosed.

Description

3D cell culture biosensor and its manufacturing method {3D CELL CULTURE BIOSENSOR AND MANUFACTURING METHOD THEREOF}

The present invention relates to a three-dimensional cell culture biosensor and its manufacturing method. Specifically, the present invention relates to a biosensor that cultivates cells three-dimensionally, measures impedance by location, and simultaneously monitors temperature.

Recently, regulations on animal experimentation have received attention as a social problem. And the importance of new methods to replace animal experiments is expanding.

Therefore, studies of culturing artificial 3D organs using 3D printing technology are developing as a method to replace animal experiments.

Electrical Cell-substrate Impedance System (ECIS) and Electrical Impedance Spectroscopy (EIS) are the most widely used methods in the field of sensors for cell condition monitoring. These methods have advantages such as low cost, non-invasiveness, and quickly monitored results. Many studies on sensors have been conducted using the above ECIS and EIS methods.

In the case of the conventional biosensor measuring impedance, cells were cultured on the deposited circuit and the reaction of the cell-drug was observed through the change of impedance and resistance through the deposited circuit. However, this method can measure only the cross-sectional (two-dimensional) characteristics of cells by measuring between the deposited circuits.

In general, a method of monitoring impedance alone has been studied as a method of monitoring the cell culture state and cell-drug response. It has been receiving much attention in its potential as a way to monitor cell culture status and drug response in a non-destructive way. In addition, impedance monitoring is an electrical measurement method that measures changes in three variables of resistance, capacitance, and inductance according to cell culture. However, it is insufficient to precisely monitor the growth process and drug response of complex cells with only electrical properties. For example, in the case of an exothermic/endothermic reaction that does not involve a change in impedance, observation is difficult. Therefore, if the change in impedance and temperature (thermal reaction) can be monitored at the same time, it can occupy a favorable position for studying the characteristics of cells.

International Publication No. WO 2015163617A1

The problem to be solved by the present invention is to provide a 3D cell culture biosensor and a manufacturing method thereof.

The problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

A three-dimensional cell culture biosensor according to an aspect of the present invention for solving the above-described problem is a temperature sensor substrate including a temperature sensor, the temperature sensor interposed therebetween, and perpendicular to the temperature sensor substrate in a direction facing each other. And a cell culture well arranged to include the plurality of impedance sensor chips and the temperature sensor and the plurality of impedance sensor chips therein, and the cells using the temperature sensor and the impedance sensor chip. The impedance and temperature change of the cells cultured in the culture well are measured.

In addition, a pattern for measuring impedance is formed on the surface of the impedance sensor chip, and the pattern may include a plurality of electrode pads.

In addition, the impedance sensor chip may further include a wire connecting the plurality of electrode pads and an impedance measurement device.

In addition, the plurality of electrode pads includes eight electrode pads continuously arranged to each other, and the eight electrode pads include a first electrode pad and an eighth electrode pad electrically connected to each other, and a second electrode pad electrically connected to each other. And a seventh electrode pad, a third electrode pad electrically connected to each other, and a fourth electrode pad and a fifth electrode pad electrically connected to each other.

In addition, the impedance sensor chip, by using a wire connected to any two electrode pads that are not electrically connected to each other of the eight electrode pads, by measuring the impedance change between the two electrode pads are not electrically connected , Impedance of the cultured cells at different locations can be measured.

In addition, further comprising a control unit for adjusting the interval between the plurality of electrode pads, the control unit, by using at least one sensor to collect information about the type and culture environment of the cultured cells, the collected information Based on the measurement accuracy of the impedance sensor chip, the gap between the plurality of electrode pads may be adjusted according to the determined accuracy.

In addition, the temperature sensor includes a platinum wire, and the temperature sensor substrate further includes a wire connecting the platinum wire and a resistance measuring device to measure a change in resistance of the platinum wire according to a temperature change of the cultured cell. , The temperature sensor may measure the temperature of the cultured cell from the resistance of the platinum wire by utilizing a temperature coefficient of resistance (TCR).

A 3D cell culture biosensor according to another aspect of the present invention for solving the above-described problem, a substrate including a temperature measuring unit composed of a platinum wire and an electrode connected to the temperature measuring unit, and the temperature measuring unit Cell culture in which the two impedance sensor chips and the temperature measuring unit and the two impedance sensor chips each including a plurality of platinum electrodes are disposed to be disposed therein and vertically disposed in the direction facing each other. PDMS wells for use, and measuring the impedance and temperature change of the cells cultured in the cell culture wells using the temperature measuring unit and the impedance sensor chip, the plurality of platinum included in the two impedance sensor chips The electrode is used to measure the three-dimensional impedance of the cultured cells at different locations in the cell culture well.

A 3D cell culture biosensor according to another aspect of the present invention for solving the above-described problem, a substrate comprising a temperature measuring unit composed of a metal wire and an electrode connected to the temperature measuring unit, and the temperature measuring unit Put in, and two impedance sensor chips arranged vertically with the substrate in the direction facing each other, PDMS well for cell culture arranged to include the temperature measuring unit and the two impedance sensor chips therein, and the electrode of the substrate A resistance measurement device connected to a wire and an impedance measurement device connected to the impedance sensor chip and a wire, wherein the resistance measurement device measures the resistance of the metal wire and utilizes a TCR from the measurement result to culture the cell. The temperature change of the cells cultured in the well is measured, and the impedance measurement device measures the impedance change of the cells cultured in the cell culture well using the impedance sensor chip.

A method of manufacturing a 3D cell culture biosensor according to an aspect of the present invention for solving the above-described problem is a step of connecting a temperature sensor to a temperature sensor substrate, with the temperature sensors interposed therebetween, and in a direction facing each other. Connecting a plurality of impedance sensor chips vertically to a temperature sensor substrate, placing cell culture wells to include the temperature sensor and the plurality of impedance sensor chips therein, and electrodes and resistance measuring devices connected to the temperature sensors And connecting the electrodes included in the plurality of impedance sensor chips and the impedance measuring device with wires.

Other specific matters of the present invention are included in the detailed description and drawings.

According to the disclosed embodiment, it is possible to obtain advantageous results in studying the characteristics of a cell by simultaneously monitoring the impedance and temperature, as well as monitoring the three-dimensional change in impedance and temperature.

The effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the following description.

1 is a diagram illustrating a 3D cell culture biosensor according to an embodiment.
2 is a view specifically showing a substrate according to an embodiment.
3 is a perspective view of a substrate according to an embodiment.
4 is a diagram illustrating a method of manufacturing an impedance sensor chip according to an embodiment.
5 is a view showing a method of manufacturing a temperature sensor substrate according to an embodiment.
6 is a view showing a method of manufacturing a 3D cell culture biosensor according to an embodiment.
7 is a diagram illustrating an impedance sensor chip according to an embodiment.
8 is a diagram illustrating an example in which a sensor is connected to an impedance measuring device.
9 is a diagram illustrating an example in which a sensor is connected to a temperature measuring device.

Advantages and features of the present invention, and methods for achieving them will be clarified with reference to embodiments described below in detail together with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below, but may be implemented in various different forms, and only the present embodiments allow the disclosure of the present invention to be complete, and are common in the technical field to which the present invention pertains. It is provided to fully inform the skilled person of the scope of the present invention, and the present invention is only defined by the scope of the claims.

The terminology used herein is for describing the embodiments and is not intended to limit the present invention. In the present specification, the singular form also includes the plural form unless otherwise specified in the phrase. As used herein, “comprises” and/or “comprising” does not exclude the presence or addition of one or more other components other than the components mentioned. Throughout the specification, the same reference numerals refer to the same components, and “and/or” includes each and every combination of one or more of the components mentioned. Although "first", "second", etc. are used to describe various components, it goes without saying that these components are not limited by these terms. These terms are only used to distinguish one component from another component. Therefore, it goes without saying that the first component mentioned below may be the second component within the technical spirit of the present invention.

Unless otherwise defined, all terms (including technical and scientific terms) used in the present specification may be used as meanings commonly understood by those skilled in the art to which the present invention pertains. In addition, terms defined in the commonly used dictionary are not ideally or excessively interpreted unless explicitly defined.

The term "part" or "module" as used in the specification refers to a hardware component such as software, FPGA, or ASIC, and "part" or "module" performs certain roles. However, "part" or "module" is not meant to be limited to software or hardware. The "unit" or "module" may be configured to be in an addressable storage medium or may be configured to reproduce one or more processors. Thus, as an example, "part" or "module" means components, processes, functions, attributes, such as software components, object-oriented software components, class components and task components. Includes procedures, subroutines, segments of program code, drivers, firmware, microcode, circuitry, data, database, data structures, tables, arrays and variables. The functionality provided within components and "parts" or "modules" can be combined into a smaller number of components and "parts" or "modules" or into additional components and "parts" or "modules" Can be further separated.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

1 is a diagram illustrating a 3D cell culture biosensor according to an embodiment.

Referring to FIG. 1, the 3D cell culture biosensor 100 includes a well 110 for cell culture, a substrate 110 on which sensors for measuring impedance and temperature of a cultured cell and changes thereof, and a cell are cultured. Kit 130 in which analysis and measurement devices for analyzing and processing information received from the sensor and the culture and measurement unit 120 including a sensor for measuring the temperature and impedance of the well and the cells cultured therein are arranged. Includes.

Each specific configuration will be described later with reference to the drawings.

2 is a view specifically showing a substrate according to an embodiment.

Referring to FIG. 2, a culture and measurement unit 120 is disposed on the substrate 110.

In one embodiment, the temperature sensor 400 is disposed on the substrate 110. The type of the temperature sensor 400 is not limited, but according to an embodiment illustrated in FIG. 2, the temperature sensor 400 measures a change in temperature using a resistance, and a metal wire for this is disposed on the substrate 110 Can be. For example, the metal wire may be a platinum or chrome wire, but is not limited thereto.

In addition, electrodes 112 and 114 connected from the temperature sensor 400 may be disposed on the substrate 110.

In addition, a plurality of impedance sensor chips 200 and 300 disposed vertically to the substrate 110 in a direction facing each other may be disposed on the substrate 110 with the temperature sensor 400 interposed therebetween. The number of the plurality of impedance sensor chips 200 and 300 is not limited, but according to the embodiment illustrated in FIG. 2, two impedance sensor chips 200 and 300 may be used.

The impedance sensor chips 200 and 300 are used for impedance monitoring, and impedance monitoring is an electrical measurement method performed by measuring changes in three variables of resistance, capacitance, and inductance according to cell culture. .

In addition, a cell culture well 500 may be disposed on the substrate 110 to include the temperature sensor 400 and the plurality of impedance sensor chips 200 and 300 therein. The material of the well 500 is not limited, but in one embodiment, a PDMS well may be used.

The temperature chip, which is the substrate for the well 500, may use various materials such as silicon, silicon membrane, silicon oxide, silicon nitride, slide glass, and fibroin coated on both sides depending on the cell culture method. You can.

In one embodiment, a substrate may be fabricated using a silicon oxide structure to improve the sensitivity and reaction rate of the temperature sensor.

In one embodiment, cells are injected into the well 500 and a culture solution is injected for culture. Over time, cells are cultured in each sensor and in empty spaces between the sensors, so that the impedance gradually changes. The change of the impedance over time may utilize a commercial impedance analyzer, or, if necessary, a small chip with a function of measuring impedance to measure it. In addition, when a temperature change occurs in the sensor part according to the cell culture and drug reaction, it can be measured by a temperature sensor under the temperature insulating layer.

The 3D cell culture biosensor 100 measures impedance and temperature changes of cells cultured in a cell culture well using the temperature sensor 400 and the impedance sensor chips 200 and 300.

3 is a perspective view of a substrate according to an embodiment.

Referring to FIG. 3, an impedance sensor chip 200 and 300, a temperature sensor 400 and electrodes 112 and 114 connected to the temperature sensor 400 are disposed on the substrate 110 as shown in FIG. 2.

In addition, patterns 210 for impedance measurement are also disposed on the impedance sensor chips 200 and 300. Specifically, a pattern 210 for impedance measurement is formed on the surfaces of the impedance sensor chips 200 and 300, and the pattern is composed of a plurality of electrode pads. The specific configuration of the pattern 210 will be described later.

4 is a diagram illustrating a method of manufacturing an impedance sensor chip according to an embodiment.

In Figure 4 (A), a silicon wafer substrate is prepared. The substrate may be composed of other materials having biocompatibility, and the type is not limited.

4(B) is an example in which spin coating is performed by dropping a PR (photoresist) solution on a prepared substrate, and in (C), a shadow mask is used to expose the desired area with ultraviolet light and develop after the process is developed. Expressed.

In (D) of FIG. 4, chromium and platinum (Cr, Pt) thin films are deposited through a sputtering process on the exposed pattern of (C), and (E) removes the remaining PR solution through a lift-off process. By explaining the appearance of the platinum wire. This can be replaced by a sputtering process using a normal shadow mask.

In general, when the line width of the pattern is ~ several tens of µm, it is widely used because the method using the shadow mask described in FIG. 4 is simple and easy to process, but when it is desired to form a fine pattern with a pattern line width of ~ µm or less, the PR solution And patterns are formed using exposure equipment. This is easy to use when the substrate and pattern become small.

5 is a view showing a method of manufacturing a temperature sensor substrate according to an embodiment.

In FIG. 5A, a silicon wafer substrate is prepared. (B) expresses spin coating by dropping the PR (photoresist) solution on the prepared substrate, and (C) exposes the desired area with ultraviolet light using a shadow mask, develops it, and then develops silicon with RIE equipment. This is after the wafer is dry etched.

In (D) of FIG. 5, a process in which a silicon wafer is removed is etched through KOH etching to create a space for an impedance chip, and (E) oxidizes a silicon wafer to deposit an SiO2 film on the silicon wafer. The growth process was expressed.

In (F) of FIG. 5, a process of depositing a platinum wire in the same manner as in FIG. 4 and making the resistance of the platinum wire stable by the furnace process is expressed. In (G), PDMS was coated for thermal separation of the temperature sensor.

6 is a view showing a method of manufacturing a 3D cell culture biosensor according to an embodiment.

Referring to FIG. 6, the substrate 110 including the temperature sensor 400 is first connected to the kit 130 as shown in FIG. 1 in (A), and the impedance sensor chips 200 and 300 in (B) ) Into the space created in (D) of FIG. 5 to connect to the kit. In (C), make a PDMS well in the sensor part, and in (D), finish it by wiring the pad for electrical measurement.

For example, the electrodes 112 and 114 connected to the temperature sensor 400 may be connected to the resistance measurement device 700 as illustrated in FIG. 9. Specifically, in order to measure the change in resistance of the platinum wire included in the temperature sensor 400 according to the temperature change of the cells cultured in the well 500, the electrodes 112 and 114 connected to the platinum wire and the resistance measuring device 700 are used. Can be connected by wire.

The temperature sensor module including the temperature sensor 400 and the resistance measurement device 700 may measure the temperature of cells cultured from the resistance of the platinum wire by utilizing a temperature coefficient of resistance (TCR).

TCR is defined as the change in resistance when the temperature changes by 1 degree. That is, TCR = 1/R ₀ *(dR/dT). On the other hand, if the resistance value at the reference temperature T ₀ is R ₀ , the resistance according to the subsequent increasing temperature is a function of temperature, which can be expressed by the following equation.

R(T) = R ₀ [1+ TCR*(TT ₀ )]

From the above formula, if the TCR of the substance is known, the resistance value at a specific temperature T can be known. Using this in reverse, if the resistance value measured at the unknown temperature T is R(T), the temperature T can be determined from the above equation.

In another embodiment, a temperature measuring device other than the resistance measuring device 700 may be connected to the temperature sensor 400 according to the temperature measuring method of the temperature sensor 400.

7 is a diagram illustrating an impedance sensor chip according to an embodiment.

Referring to FIG. 7, a pattern 210 is disposed on the impedance sensor chip 200, and the pattern 210 is composed of a plurality of electrodes made of a metal containing platinum or chromium.

In addition, as described above, each of the plurality of electrodes included in the impedance sensor chip 200 may be connected to the impedance measurement device 600 as illustrated in FIG. 8.

Referring to FIG. 7, the pattern 210 includes eight electrode pads continuously arranged with each other, and the eight electrode pads include first electrode pads 211 and eighth electrode pads 218 electrically connected to each other. , A second electrode pad 212 and a seventh electrode pad 217 electrically connected to each other, a third electrode pad 213 and a sixth electrode pad 216 electrically connected to each other, and a fourth electrode pad electrically connected to each other ( 214) and a fifth electrode pad 215.

In addition, the impedance sensor chip 200 measures impedance change between two electrode pads that are not electrically connected by using a wire connected to any two electrode pads that are not electrically connected to each other among the eight electrode pads. , Impedance of cells cultured at different locations can be measured.

For example, between two electrode pads that are not electrically connected to each other, it is possible to measure a change in impedance according to the growth of a cultured cell or a drug reaction.

Referring to FIG. 7, each electrode pad is spaced apart from each other. This distance should be appropriately adjusted depending on the type of cell culture and the culture environment, and is usually configured within 1 μm to 1 mm, but in special cases, it may be reduced to less than 1 μm. It can be seen that the narrower the interval, the higher the measurement accuracy of the impedance of the biosensor.

In one embodiment, the 3D cell culture biosensor 100 may further include a control unit that adjusts an interval between a plurality of electrode pads disposed on the impedance sensor chip 200.

For example, a plurality of electrode pads may be fixedly deposited on the impedance sensor chip 200, but the position and spacing may be adjusted using a power unit. The type of the power unit is not limited, and each power unit may be provided for each electrode, or power may be transmitted from one power unit to each electrode by using gears or the like, and differential power transmitted using a gear ratio may be differential. You can also adjust the enemy.

In one embodiment, the control unit collects information on the type and culture environment of the cultured cells using at least one sensor, and determines the measurement accuracy of the impedance sensor chip based on the collected information, and the determined The spacing between the plurality of electrode pads may be adjusted according to precision.

For example, the control unit may determine the precision of the impedance measurement required according to the temperature change measured by the temperature sensor 400, and accordingly, the gap between the plurality of electrode pads may be narrowed by controlling the power unit.

Similarly, when it is determined that the precision of impedance measurement is not required to be large, the control unit may widen the gap between the plurality of electrode pads by controlling the power unit.

In addition, when a separate sensor is included in the 3D cell culture biosensor 100, the control unit may determine the weight, volume, color, and movement of the cultured cell. Accordingly, the control unit may widen the gap between the plurality of electrode pads by controlling the power unit.

For example, if the cultured cells are small, there may be electrode pads that do not contact the cells among the plurality of electrode pads or have a small contact area with the cells. In this case, the control unit may control the power unit to narrow the gap between the plurality of electrode pads or shift the plurality of electrode pads to move the plurality of electrode pads to the position where the cells are cultured. For example, the control unit, based on the impedance change measured from each electrode pad, if there is an electrode with little or no impedance change, the electrode or a plurality of electrodes including the electrode until an impedance change occurs in the electrode Can be moved.

The embodiments of the present invention have been described with reference to the accompanying drawings, but a person skilled in the art to which the present invention pertains may implement the present invention in other specific forms without changing its technical spirit or essential features. You will understand. Therefore, it should be understood that the above-described embodiments are illustrative in all respects and not restrictive.

100: 3D cell culture biosensor
110: substrate
120: sensor unit

Claims (10)

A temperature sensor substrate including a temperature sensor;
A plurality of impedance sensor chips disposed vertically with the temperature sensor substrate in a direction facing each other with the temperature sensor interposed therebetween; And
And a cell culture well arranged to include the temperature sensor and the plurality of impedance sensor chips therein,
Using the temperature sensor and the impedance sensor chip to measure the impedance and temperature change of the cells cultured in the cell culture well,
An arbitrary pattern for impedance measurement is formed on the surface of the impedance sensor chip, and the pattern is composed of a plurality of electrode pads,
The impedance sensor chip,
A wire connecting the plurality of electrode pads and an impedance measurement device; Further comprising,
The plurality of electrode pads includes at least eight electrode pads continuously arranged to each other,
The eight electrode pads,
A first electrode pad and an eighth electrode pad electrically connected to each other;
A second electrode pad and a seventh electrode pad electrically connected to each other;
A third electrode pad and a sixth electrode pad electrically connected to each other; And
A fourth electrode pad and a fifth electrode pad electrically connected to each other; Including, 3D cell culture biosensor. delete delete delete According to claim 1,
The impedance sensor chip,
Using the wire connected to any two electrode pads that are not electrically connected to each other among the eight electrode pads, the impedance is measured between the two electrode pads that are not electrically connected to each other, thereby culturing the cells at different locations. A 3D cell culture biosensor measuring the impedance of cells. According to claim 1,
A control unit adjusting an interval between the plurality of electrode pads; Further comprising,
The control unit,
Collecting information about the type and culture environment of the cultured cells using at least one sensor, determining the measurement precision of the impedance sensor chip based on the collected information, and the plurality of electrodes according to the determined precision A 3D cell culture biosensor that adjusts the spacing between pads. delete A substrate including a temperature measuring unit formed of a platinum wire and an electrode connected to the temperature measuring unit;
Two impedance sensor chips disposed vertically with the substrate in a direction facing each other with the temperature measuring unit interposed therebetween and including a plurality of platinum electrodes; And
A PDMS well for cell culture, which is arranged to include the temperature measuring part and the two impedance sensor chips therein; Including,
Measuring the impedance and temperature change of cells cultured in the PDMS well for cell culture by using the temperature measuring unit and the impedance sensor chip, the cells using the plurality of platinum electrodes included in the two impedance sensor chips Measure the three-dimensional impedance of the cultured cells at different locations in the PDMS well for culture,
An arbitrary pattern for impedance measurement is formed on the surface of the impedance sensor chip, and the pattern is composed of a plurality of electrode pads,
The impedance sensor chip,
A wire connecting the plurality of electrode pads and an impedance measurement device; Further comprising,
The plurality of electrode pads includes at least eight electrode pads continuously arranged to each other,
The eight electrode pads,
A first electrode pad and an eighth electrode pad electrically connected to each other;
A second electrode pad and a seventh electrode pad electrically connected to each other;
A third electrode pad and a sixth electrode pad electrically connected to each other; And
A fourth electrode pad and a fifth electrode pad electrically connected to each other; Including, 3D cell culture biosensor. A substrate including a temperature measuring unit formed of a metal wire and an electrode connected to the temperature measuring unit;
Two impedance sensor chips disposed vertically with the substrate in a direction facing each other with the temperature measuring unit interposed therebetween;
A PDMS well for cell culture, which is arranged to include the temperature measuring part and the two impedance sensor chips therein;
A resistance measuring device connected to the electrode and the wire of the substrate; And
An impedance measuring device connected to the impedance sensor chip by a wire; Including,
The resistance measuring device measures the resistance of the metal wire, and measures the temperature change of the cells cultured in the PDMS well for cell culture by utilizing the TCR from the measurement results,
The impedance measurement device measures the impedance change of cells cultured in the PDMS well for cell culture using the impedance sensor chip,
An arbitrary pattern for impedance measurement is formed on the surface of the impedance sensor chip, and the pattern is composed of a plurality of electrode pads,
The plurality of electrode pads includes at least eight electrode pads continuously arranged to each other,
The eight electrode pads,
A first electrode pad and an eighth electrode pad electrically connected to each other;
A second electrode pad and a seventh electrode pad electrically connected to each other;
A third electrode pad and a sixth electrode pad electrically connected to each other; And
A fourth electrode pad and a fifth electrode pad electrically connected to each other; Including, 3D cell culture biosensor. Connecting a temperature sensor to the temperature sensor substrate;
Connecting the plurality of impedance sensor chips perpendicularly to the temperature sensor substrate in a direction facing each other with the temperature sensors interposed therebetween;
Placing the cell culture wells to include the temperature sensor and the plurality of impedance sensor chips therein;
Connecting an electrode connected to the temperature sensor and a resistance measuring device with a wire; And
Connecting an electrode included in the plurality of impedance sensor chips and an impedance measuring device with a wire; Including,
An arbitrary pattern for impedance measurement is formed on the surface of the impedance sensor chip, and the pattern is composed of a plurality of electrode pads,
The impedance sensor chip,
A wire connecting the plurality of electrode pads and an impedance measurement device; Further comprising,
The plurality of electrode pads includes at least eight electrode pads continuously arranged to each other,
The eight electrode pads,
A first electrode pad and an eighth electrode pad electrically connected to each other;
A second electrode pad and a seventh electrode pad electrically connected to each other;
A third electrode pad and a sixth electrode pad electrically connected to each other; And
A fourth electrode pad and a fifth electrode pad electrically connected to each other; The method of manufacturing a 3D cell culture biosensor comprising a.

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