Microfluidic chip and method for culturing single cells and sorting and extracting cell populations.

Abstract

A microfluidic chip and a method for culturing a single cell and selecting and extracting a cell group are provided. The chip (1) includes a substrate layer, a flow path layer, a liquid supply flow path (2), a liquid discharge flow path (3), a plurality of common flow paths (4), and a plurality of functional units (5). The flow path layer is located below the substrate layer. The material of the flow path layer includes at least one of silicon dioxide, spin-on glass, non-photosensitive epoxy resin, and non-photosensitive polyimide. Both ends of the common flow path (4) are connected to the liquid supply flow path (2) and the liquid discharge flow path (3), respectively. The functional unit (5) includes a single cell inlet (501), a cell culture selection chamber (502), a cell discharge chamber (503), a cell discharge port (504), and a drive member (505). The driving member (505) is used to supply power to the liquid to draw the single cell (6) introduced into the common flow channel (4) into the cell culture and selection chamber (502) and to discharge the cultured and selected target cell group from the cell outlet (504). The microfluidic chip can complete the culture of a single cell, the identification and selection of the cell group, and the discharge of the target cell group all on the chip, making experimental operations easier, reducing the risk of cross-contamination, and avoiding fluorescent interference from the flow channel layer itself.

Description

Technical Field

[0001] The present invention belongs to the fields of microfluidics, development of cell lines, selection of monoclonal antibodies, etc., and relates to a microfluidic chip and a method for culturing single cells and selecting and deriving cell populations.

Background Art

[0002] Cells are the basic units of life activities. Therefore, research at the single-cell level can reveal the laws of the development of life activities from a deeper level and is widely applied in fields such as the selection of monoclonal antibodies and the culture of cell lines. The separation of single cells is the basis and key point of single-cell research. Currently, the main methods for separating single cells are the micropipette aspiration method, the limiting dilution method, the micro-well array, and the sorting method based on microfluidics. However, the current separation methods face problems such as high operation difficulty, low efficiency, and acquisition of multiple cells, which are inconvenient for subsequent culture and analysis. In addition, when applied to the selection of monoclonal antibodies and the culture of cell lines, single cells after separation are placed on a microplate for culture, and after performing potency and phenotype analysis on the cell population, a cell population with excellent performance is often selected for large-scale culture. Therefore, the entire process requires a great deal of labor, the operation is complicated, it takes a long time, and the efficiency is low. Therefore, especially in single-cell research such as the selection of monoclonal antibodies and the development of cell lines, a research method with easy operation and high efficiency that integrates the separation and culture of single cells and the selection and derivation of cell populations is urgently required.

Summary of the Invention

Problems to be Solved by the Invention

[0003] In view of the shortcomings of the conventional technology described above, the object of the present invention is to provide a microfluidic chip and a method for culturing single cells and selecting and extracting cell populations, in order to solve the problems that processes such as cell isolation and culture and cell population selection in the conventional technology require a great deal of effort, are complicated to operate, take a long time, and are inefficient. [Means for solving the problem]

[0004] To achieve the above-mentioned objectives and other related objectives, the present invention provides an integrated microfluidic chip. The integrated microfluidic chip comprises a substrate layer, a channel layer located below the substrate layer and made of at least one of silicon dioxide, spin-on glass, non-photosensitive epoxy resin and non-photosensitive polyimide, a supply channel and an discharge channel located in the channel layer and provided at intervals, a plurality of common channels located in the channel layer and provided at intervals, the common channels having both ends communicating with the supply channel and the discharge channel, respectively, and a plurality of functional units including a single-cell inlet, a cell culture sorting chamber, a cell exit chamber, a cell exit port and a drive member, wherein the single-cell inlet is located in the substrate layer and communicates with the common channels, The invention includes a functional unit in which a cell culture sorting chamber, a cell discharge chamber, and a cell outlet are all located in the flow channel layer, and both ends of the cell culture sorting chamber are in communication with the common flow channel and the cell discharge chamber, respectively, the cell outlet is located below the cell discharge chamber and is in communication with the cell discharge chamber, the drive member is located in the cell discharge chamber and faces the cell outlet, and the drive member supplies power to a liquid to draw single cells introduced into the common flow channel through the single cell inlet into the cell culture sorting chamber, and to discharge the target cell group cultured and sorted in the cell culture sorting chamber from the cell outlet.

[0005] Selectively, the fluid supply channel, the discharge channel, the common channel, the cell culture sorting chamber, and the cell discharge chamber all penetrate the surface of the channel layer facing the substrate layer, exposing the substrate layer. The drive member is coupled to the surface of the substrate layer.

[0006] Selectively, the material of the substrate layer includes silicon.

[0007] Selectively, the extension direction of the liquid supply channel and the extension direction of the liquid discharge channel are parallel.

[0008] Selectively, the extension direction of the common channel and the extension direction of the liquid supply channel are perpendicular.

[0009] Selectively, the thickness of the cell culture sorting chamber is set to accommodate only a single layer of cells.

[0010] Selectively, in the plane where the flow channel layer is located, and in a direction perpendicular to the direction in which the cell culture sorting chamber faces the cell discharge chamber, the width of the cell culture sorting chamber is greater than the width of the cell discharge chamber.

[0011] Selectively, the single-cell entry port is used to receive single cells ejected from a single-cell printing chip.

[0012] Selectively, the single-cell printing chip includes a thermal bubble printing chip.

[0013] Selectively, the drive member includes one of a heating film, a piezoelectric nozzle, a PDMS microvalve, a solenoid valve, and a peristaltic pump.

[0014] Selectively, the number of the functional units ranges from 10 to 10,000.

[0015] The present invention further provides a method for culturing single cells and sorting and extracting cell populations. The method includes providing a microfluidic chip as described above, injecting a single cell into the common channel through the single cell inlet, drawing the single cell injected into the common channel into the cell culture sorting chamber by causing the liquid to flow with the drive member after the cell has settled naturally, culturing the cells in the cell culture sorting chamber for a predetermined time, and then introducing a sorting reagent into the cell culture sorting chamber through the liquid supply channel to identify and sort the target cell population, and transferring the target cell population into a predetermined container through the cell outlet.

[0016] Selectively, before injecting a single cell into the common channel through the single-cell inlet, the cell culture medium is flowed into the integrated microfluidic chip to remove air bubbles.

[0017] After selectively drawing single cells injected into the common channel into the cell culture sorting chamber, the cell culture medium is again introduced to culture the cells.

[0018] Selectively, the cell culture medium is introduced using a syringe pump.

[0019] After selectively introducing a selection reagent into the cell culture sorting chamber, the target cell population is selected by performing fluorescence imaging characterization of the cell population in the cell culture sorting chamber.

[0020] The method for selectively culturing single cells and selecting and extracting cell populations is used for selecting cell populations of monoclonal antibodies. [Effects of the Invention]

[0021] As described above, the present invention provides a microfluidic chip used for the separation and culture of single cells, as well as the selection and derivation of target cell populations. Since all of the culture of single cells, the identification and selection of cell populations, and the derivation of target cell populations can be completed on the chip, the cell throughput is improved, the experimental operation is simplified, and the usage amount of reagents and the risk of potential cross-contamination are reduced. Moreover, by combining the thermal bubble printing technology, the whole process becomes easier to control, and it is simple and highly efficient. In addition, in the microfluidic chip of the present invention, a silicon dioxide layer, a spin-on glass layer, a non-photosensitive epoxy resin material layer, or a non-photosensitive polyimide material layer is used in the flow channel layer instead of the conventional dry film, so that the fluorescence interference of the flow channel layer itself can be avoided.

Brief Description of the Drawings

[0022] [Figure 1] FIG. 1 shows a plan layout diagram of the flow channels in the microfluidic chip of the present invention. [Figure 2] FIG. 2 shows a longitudinal layout diagram of the flow channels of the functional units in the microfluidic chip of the present invention. [Figure 3] FIG. 3 shows a flowchart of a method for culturing single cells and selecting and deriving cell populations in the present invention.

Embodiments for Carrying Out the Invention

[0023] Hereinafter, embodiments of the present invention will be described by specific specific examples. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. Furthermore, the present invention may be implemented or applied by other different specific embodiments. Also, for each detailed matter in this specification, various supplements or modifications may be made on the premise of not departing from the spirit of the present invention based on different viewpoints and applications.

[0024] Refer to FIGS. 1 to 3. It should be noted that the drawings provided in this embodiment only schematically illustrate the basic idea of the present invention. The drawings only show the assemblies related to the present invention, and are not described based on the number, shape and size of the assemblies actually implemented. The form, number and ratio of each assembly actually implemented may be arbitrarily changed, and the layout and form of the assemblies may become more complex.

Embodiment

[0025] In this embodiment, a microfluidic chip including a substrate layer, a channel layer, a liquid supply channel, a liquid discharge channel, a plurality of common channels and a plurality of functional units is provided. The channel layer is located below the substrate layer.

[0026] Specifically, the material of the channel layer includes at least one of silicon dioxide, spin-on glass (abbreviated as SOG, a spin-coating material of polyoxysilane), non-photosensitive epoxy resin and non-photosensitive polyimide. In the channel layer of the microfluidic chip in the present invention, instead of the conventional dry film, for example, a non-photosensitive material layer such as a silicon dioxide layer, a spin-on glass layer, a non-photosensitive epoxy resin material layer or a non-photosensitive polyimide material layer is used, so that the problem of fluorescence interference of the channel layer itself can be avoided.

[0027] As an example, the material of the substrate layer includes silicon, and desired circuit elements and circuits can be fabricated on this layer. In addition, since a silicon dioxide layer, a spin-on glass layer, a non-photosensitive epoxy resin material layer or a non-photosensitive polyimide material layer is used for the channel layer, not only can the problem of fluorescence interference of the channel layer itself be avoided, but also good bonding with the silicon substrate can be achieved. Also, it becomes easy to process the channel layer on the silicon substrate using micro-nano technology to form a desired channel. For example, first, a first channel layer is formed on the substrate layer, and a desired channel is obtained by patterning the first channel layer. Next, a second channel layer is formed on the first channel layer to block the channels in the first channel layer.

[0028] As an example, refer to Figures 1 and 2. Figure 1 shows a planar layout of the flow channels in the microfluidic chip 1, and Figure 2 shows a longitudinal layout of the flow channels of the functional unit in the microfluidic chip.

[0029] Specifically, the liquid supply channel 2 and the discharge channel 3 are both located in the channel layer and are provided at intervals. Furthermore, the multiple common channels 4 are located in the channel layer and are provided at intervals. Both ends of the common channels 4 are in communication with the liquid supply channel 2 and the discharge channel 3, respectively. The multiple functional units 5 are arranged in an array. Each functional unit 5 includes a single-cell inlet 501, a cell culture sorting chamber 502, a cell outlet 503, a cell outlet 504, and a drive member 505. The single-cell inlet 501 is located in the substrate layer and is in communication with the common channels 4. The cell culture sorting chamber 502, the cell outlet 503, and the cell outlet 504 are all located in the channel layer. Furthermore, both ends of the cell culture sorting chamber 502 are in communication with the common channels 4 and the cell outlet 503, respectively. The cell outlet 504 is located below the cell outlet chamber 503 and is in communication with the cell outlet chamber 503. The drive member 505 is located at the bottom of the cell outlet chamber 503 and faces the cell outlet 504. The drive member 505 is used to supply power to the liquid, thereby drawing single cells 6 introduced into the common channel 4 through the single cell inlet 501 into the cell culture sorting chamber 502, and to discharge the target cell group cultured and sorted in the cell culture sorting chamber 502 from the cell outlet 504.

[0030] As an example, the liquid supply channel 2, the discharge channel 3, the common channel 4, the cell culture sorting chamber 502, and the cell discharge chamber 503 all penetrate the surface of the channel layer facing the substrate layer, exposing the substrate layer. In addition, the drive member 505 is coupled to the surface of the substrate layer.

[0031] Specifically, the functional unit 5 has the functions of separating and culturing single cells, as well as sorting and extracting cell populations. If necessary, integrating a large number of the functional units 5 into the integrated microfluidic chip ensures high throughput for single-cell culture and sorting. For example, the number of functional units 5 ranges from 10 to 10,000, such as 1,000, 2,000, 5,000, etc.

[0032] As an example, Figure 1 shows the liquid flow path with dashed arrows. The inlet of the liquid supply channel 2 and the outlet of the liquid discharge channel 3 can be adjusted as needed, for example, to one end of the channel or a predetermined position in the center of the channel, although these are not shown in the figure. In addition, the culture medium and reagents may be operated by external power (e.g., a syringe pump) to ensure cell culture and sorting.

[0033] As an example, as shown in Figure 1, the extension direction of the fluid supply channel 2 and the extension direction of the fluid discharge channel 3 are parallel, and the extension direction of the common channel 4 and the extension direction of the fluid supply channel 2 are perpendicular. This allows the multiple functional units 5 to be arranged in a regular rectangular pattern, which is convenient for coordination with the printing nozzle of the single-cell printing chip and also convenient for coordination with the cell receiving device. In other embodiments, the communication method of the fluid supply channel 2, the fluid discharge channel 3 and the common channel 4 may be adjusted as needed, and the invention is not limited to this embodiment.

[0034] As an example, as shown in Figure 2, the thickness of the cell culture sorting chamber 502 is set to accommodate only a single layer of cells. For example, the thickness of the cell culture sorting chamber 502 may be equal to the size of the cells to be sorted. This ensures that the cells are arranged in a single layer during the cell culture process, which is convenient for subsequent cell counting and fluorescence analysis.

[0035] As an example, as shown in Figure 1, in the plane where the flow channel layer is located, and in a direction perpendicular to the direction in which the cell culture sorting chamber 502 faces the cell discharge chamber 503, the width of the cell culture sorting chamber 502 is greater than the width of the cell discharge chamber 503. This is convenient for the sorted cells to enter and be discharged into the cell discharge chamber 503 in sequence.

[0036] As an example, the drive member 505 may include any of the following: a heating film, a piezoelectric nozzle, a PDMS (polydimethylsiloxane) microvalve, a solenoid valve, and a peristaltic pump. In this embodiment, it is preferable to use a heating film for the drive member 505 to constitute a thermal bubble nozzle together with the cell outlet chamber 503 and the cell outlet 504. The thermal bubble nozzle is manufactured using micro- and nano-fabrication technology and integrated into the bottom of the microchannel. The thermal bubble nozzle utilizes the instantaneous high temperature of the heating film to vaporize the liquid above, generating bubbles to cause the liquid to flow and discharge from the nozzle. Subsequent liquid is then supplied by capillary action, providing power to continuously flow the liquid. The thermal bubble nozzle is controlled by the circuitry below.

[0037] As an example, as shown in Figure 2, the single-cell inlet 501 is used to receive single cells 6 ejected from the single-cell printing chip 7. The position of the single-cell inlet 501 is such that the settling position of the single cells is in the entrance area of ​​the cell culture sorting chamber 502, so that after the single cells enter the common channel from the single-cell inlet 501, they are accurately driven into the cell culture sorting chamber 502 by the driving action of the driving member 505. The single-cell printing chip 7 may be the single-cell printing chip disclosed in Chinese Patent Application Publication CN108330065A, or any other suitable single-cell printing chip. This enables the separation of single cells and their introduction into the single-cell inlet 501. In this embodiment, it is preferable to use a single-cell printing chip that includes a thermal bubble printing chip that minimizes cell damage and allows for the efficient and gentle introduction of single cells into the single-cell inlet 501.

[0038] The integrated microfluidic chip of this invention can be used for the isolation and culture of single cells, as well as the selection and extraction of target cell populations. This allows single-cell culture, cell population identification and selection, and target cell population extraction to all be completed on the chip, improving cell throughput, simplifying experimental procedures, and reducing reagent usage and the risk of potential cross-contamination. Furthermore, combining this with thermal bubble printing technology makes the entire process even easier to control, resulting in a convenient and highly efficient operation. [Examples]

[0039] This embodiment provides a method for culturing single cells and for selecting and extracting cell populations. A flowchart of this method is shown in Figure 3. The method includes the following steps.

[0040] Step S1 provides the integrated microfluidic chip described in Example 1, and injects a single cell into the common channel through the single cell inlet.

[0041] Step S2 involves drawing the single cells injected into the common channel into the cell culture sorting chamber by causing the liquid to flow using the driving member after the cells have settled naturally.

[0042] Step S3 involves culturing cells in the cell culture sorting chamber for a predetermined time, and then introducing a sorting reagent into the cell culture sorting chamber through the supply channel to identify and sort the target cell population.

[0043] Step S4 involves transferring the target cell population into a predetermined container through the cell exit port.

[0044] As an example, in step S1, before injecting a single cell into the common channel from the single cell inlet, the cell culture medium is poured into the integrated microfluidic chip to remove air bubbles.

[0045] As an example, in step S2, single cells injected into the common channel are drawn into the cell culture sorting chamber, and then cell culture medium is again introduced to diffuse and introduce nutrients into the cell culture sorting chamber, thereby culturing the cells. In this embodiment, after all single cells have entered the corresponding cell culture sorting chamber, the cell culture medium is introduced using a syringe pump.

[0046] As an example, in step S3, after introducing the selection reagent into the cell culture selection chamber, the target cell population is selected by performing fluorescence image characterization of the cell population in the cell culture selection chamber. The selection reagent may be a phenotypic antibody or other target, and is specifically determined according to the cells to be selected. In this embodiment, after the cells have been cultured in the cell culture selection chamber for several days, the phenotypic antibody or other target is injected into a chip, and the target cell population with superior performance is selected based on the fluorescence characteristics of the cell population.

[0047] As an example, in step S4, the target cell population is preferably transferred to a predetermined container using a thermal bubble printing method, and then cultured or analyzed. The thermal bubble printing method has the advantages of causing minimal cell damage, a fast reaction time, strong driving force, ease of control, and ease of integration and miniaturization, thus ensuring that the entire process can be carried out easily and efficiently.

[0048] As an example, the method for culturing single cells and selecting and extracting cell populations in this embodiment can be used for selecting monoclonal antibody cell populations or other types of cell populations. [Examples]

[0049] In this example, monoclonal cell lines are sorted using the microfluidic chip from Example 1. First, cell culture medium is poured into the chip and air bubbles are removed. Next, the flow of culture medium is stopped, and the transfected cells are passed through a single-cell printing chip to be placed in the single-cell inlet of the chip in a single-cell form. Then, the operation of the thermal bubble nozzle is controlled by the lower circuit to allow the single cells to enter the cell culture sorting chamber. The height of the cell culture sorting chamber may be adjusted according to the size of the cells used to obtain a cell population in a single layer. Next, the culture medium is poured in again, and the cells are cultured until they form a certain number of colonies. Then, the potency of the antibodies secreted by each cell population is characterized by injecting the relevant fluorescent antibody into the reagent inlet. After that, target cell populations are sorted as needed. In this case, the thermal bubble nozzle at the cell outlet is used again to transfer the target cell population into a predetermined container, completing the sorting of monoclonal antibody cell populations.

[0050] In summary, the present invention provides an integrated microfluidic chip used for the isolation and culture of single cells and the selection and extraction of target cell populations. Because the chip allows for the completion of single-cell culture, cell population identification and selection, and target cell population extraction all on the chip itself, cell throughput is improved, experimental procedures are simplified, and reagent usage and the risk of potential cross-contamination are reduced. Furthermore, by combining this with thermal bubble printing technology, the entire process becomes even easier to control, resulting in a convenient and highly efficient operation. In addition, the microfluidic chip of the present invention uses a silicon dioxide layer, a spin-on glass layer, a non-photosensitive epoxy resin material layer, or a non-photosensitive polyimide material layer in the channel layer instead of a conventional dry film, thus avoiding fluorescence interference from the channel layer itself. Therefore, the present invention effectively overcomes various drawbacks of the prior art and possesses high industrial value.

[0051] The above embodiments are merely illustrative examples illustrating the principles and effects of the present invention and are not intended to limit it. Those familiar with the art may supplement or modify the above embodiments without departing from the spirit and scope of the invention. Accordingly, any equivalent supplement or modification that a person skilled in the art could complete without departing from the spirit and technical concept disclosed herein remains within the scope of the claims of the present invention. [Explanation of Symbols]

[0052] 1 Microfluidic Chip 2 Liquid supply channel 3. Discharge channel 4 Common channel 5 Function Units 501 Single-cell entry port 502 Cell Culture Sorting Room 503 Cell extraction room 504 Cell outlet 505 Drive Member 6 Single cell 7 Single-cell printing chip S1~S4 Steps

Claims

1. A microfluidic chip comprising a substrate layer, a channel layer, a liquid supply channel, a liquid discharge channel, a plurality of common channels, and a plurality of functional units, The flow channel layer is provided following the substrate layer along the fluid flow path, and its material comprises at least one of silicon dioxide, spin-on glass, non-photosensitive epoxy resin, and non-photosensitive polyimide. The liquid supply channel and the liquid discharge channel are located in the channel layer and are provided at intervals from each other. The aforementioned multiple common channels are located in the channel layer, are spaced apart, and both ends are in communication with the liquid supply channel and the liquid discharge channel, respectively. The plurality of functional units include a single-cell inlet, a cell culture sorting chamber, a cell outlet, a cell outlet, and a drive member. The single-cell inlet is located in the substrate layer and communicates with the common channel, the cell culture sorting chamber, the cell outlet, and the cell outlet are all located in the channel layer, and both ends of the cell culture sorting chamber communicate with the common channel and the cell outlet, respectively, and the cell outlet communicates with the cell outlet. The drive member is located in the cell outlet and faces the cell outlet. The drive member supplies power to the liquid to draw single cells introduced into the common channel through the single-cell inlet into the cell culture sorting chamber, and to discharge the target cell population cultured and sorted in the cell culture sorting chamber from the cell outlet. A microfluidic chip characterized by the following features.

2. The microfluidic chip according to claim 1, characterized in that the liquid supply channel, the discharge channel, the common channel, the cell culture sorting chamber, and the cell discharge chamber all penetrate the surface of the channel layer facing the substrate layer to expose the substrate layer, and the drive member is coupled to the surface of the substrate layer.

3. The microfluidic chip according to claim 1, characterized in that the material of the substrate layer includes silicon.

4. The microfluidic chip according to claim 1, characterized in that the extension direction of the liquid supply channel and the extension direction of the liquid discharge channel are parallel.

5. The microfluidic chip according to claim 4, characterized in that the extension direction of the common channel and the extension direction of the liquid supply channel are perpendicular.

6. The microfluidic chip according to claim 1, characterized in that the thickness of the cell culture sorting chamber is provided to accommodate only a single layer of cells.

7. The microfluidic chip according to claim 1, characterized in that, in the plane where the flow channel layer is located and in a direction perpendicular to the direction in which the cell culture sorting chamber faces the cell discharge chamber, the width of the cell culture sorting chamber is greater than the width of the cell discharge chamber.

8. The microfluidic chip according to claim 1, characterized in that the single-cell inlet is used to receive a single cell ejected from a single-cell printing chip.

9. The microfluidic chip according to claim 8, characterized in that the single-cell printing chip includes a thermal bubble printing chip.

10. The microfluidic chip according to claim 1, characterized in that the drive member includes any one of a heating film, a piezoelectric nozzle, a PDMS microvalve, a solenoid valve, and a peristaltic pump.

11. The microfluidic chip according to claim 1, characterized in that the number of the aforementioned functional units is in the range of 10 to 10,000.

12. A microfluidic chip according to any one of claims 1 to 11, comprising the steps of injecting a single cell into the common channel from the single cell inlet, After the cells have settled naturally, the driving member causes the liquid to flow, thereby drawing the single cells injected into the common channel into the cell culture sorting chamber, which is included in one of the plurality of functional units that communicate with the common channel. After culturing cells in the cell culture sorting chamber for a predetermined time, a sorting reagent is introduced into the cell culture sorting chamber through the supply channel to identify and sort the target cell population cultured in one of the multiple functional units. A method for culturing single cells and selecting and discharging cell populations, characterized by comprising the step of transferring a target cell population into a predetermined container through the cell outlet.

13. The method for culturing single cells and selecting and extracting cell populations according to claim 12, characterized in that, before injecting a single cell into the common channel from the single cell inlet, the cell culture medium is flowed into the microfluidic chip to remove air bubbles.

14. The method for culturing single cells and selecting and releasing a group of cells according to claim 13, characterized in that single cells injected into the common channel are drawn into the cell culture sorting chamber, and then cell culture medium is again introduced to culture the cells.

15. The method for culturing single cells and selecting and discharging cell populations according to claim 14, characterized in that the cell culture medium is introduced using a syringe pump.

16. The method for culturing single cells and selecting and extracting cell populations according to claim 12, characterized in that, after introducing a selection reagent into the cell culture selection chamber, the target cell population is selected by performing fluorescence image characteristic evaluation of the cell population in the cell culture selection chamber.

17. The method for culturing a single cell and selecting and extracting a cell population is characterized in that it is used for selecting a cell population of a monoclonal antibody, which is obtained by culturing a single cell and consists of multiple antibody cells, as described in claim 12.

Images