Patent Application
20240158729
The present disclosure relates to a platform which delivers intracellular materials, and more particularly, to a platform capable of delivering materials within cells by the formation of a vortex.
Intracellular material transfer is one of the most basic experiments in cell engineering, and materials are usually delivered using carriers or by creating nanopores in cell membranes/nuclear membranes. Virus- or Lipofectamine-based carrier techniques can deliver materials with high efficiency when optimized, but have problems such as safety, slow delivery speed, labor/cost-intensive carrier preparation process, and low reproducibility.
On the contrary, methods of making nanopores by applying energy to cell membranes, for example, technologies such as electroporation or microneedle, have the advantage of being able to deliver relatively various materials to various cells. However, low cell viability due to the invasiveness of the methods, denaturation of a delivery material, and low throughput have been pointed out as major limitations. In order to solve such problems, the use of microfluidic devices capable of processing a large amount of cells is remarkable. Representatively, there is a platform that creates a narrow constriction or bottleneck in the microchannels and creates nanopores in the cell membrane through physical deformation of the cells when the cells pass through the narrow constriction or bottleneck. However, this approach has major disadvantages such as clogging of the narrow constriction or bottleneck itself, inconsistent material delivery efficiency, and the like during the experiment.
For example, US Patent No. 2014-0287509 (hereinafter, the conventional technology) discloses a technique of inducing cell transformation by directly flowing cells through a channel having a bottleneck structure, thereby applying pressure to the cells. However, in this case, since the cells progress at a non-uniform speed, the rate at which the cells are transformed is not constant, and thus the efficiency of material delivery is reduced. In addition, since cells are delivered only by a method due to diffusion, the efficiency of material delivery is fundamentally low and there is a problem in that it is difficult to deliver nucleic acids into the nucleus. Therefore, it is urgent to develop an innovative next-generation intracellular material delivery platform that can deliver various materials uniformly and with high efficiency into the cells while maintaining the high processing function of the microfluidic device.
An object of the present disclosure is to provide a platform capable of delivering materials into cells by generating vortices in microchannels.
According to one aspect of the present disclosure, one embodiment of the present disclosure may include: a first channel through which a fluid containing cells and a delivery material flows while having a flow; and a second channel and a third channel which are connected to the first channel at an angle so that the fluid containing the cells and the delivery material flows while having a flow; wherein the fluid forms at least one of a collision region and a vortex region of the fluid in at least one of the first channel, the second channel, and the third channel.
In one embodiment, the cross section of at least one of the first to third channels may be formed as a rectangle having a short axis and a long axis, the short axis is provided as a vertical plane, and the long axis is provided as a horizontal plane.
In one embodiment, the material delivery platform may further include: a first supply part supplying a first supply fluid containing at least one of cells and a delivery material; and a second supply part supplying a second supply fluid, wherein the first supply fluid may be supplied to the first channel through a first supply part channel, the second supply fluid may be supplied to the first channel through second supply part channels, one or more of the second supply part channels may be formed, and the second supply part channels may supply the second fluid from both sides of the first supply part channel.
In one embodiment, the Reynolds number of the first supply part channel to the Reynolds number of the second supply part channel may be 2:1 to 1:3.
In one embodiment, the fluid in the first channel may include the first supply fluid and the second supply fluid, the first supply fluid may flow by being flowed from the center of the long axis in the first channel, the second supply fluid may flow by being flowed from both sides of the first supply fluid and flow by being flowed from both ends of the long axis in the first channel, and the first supply fluid and the second supply fluid may be mixed in at least one of the second channel and the third channel.
In one embodiment, the vortex region may be one in which a straight line-type flow of the fluid is temporarily stagnant.
In one embodiment, the delivery material may be at least one of nucleic acids, proteins, fluorescent dyes, quantum dots, carbon nanotubes, antigens, ribonucleoproteins, genetic scissors, polymers, and nanoparticles.
In one embodiment, the second channel and the third channel may be formed symmetrically or asymmetrically with respect to the first channel.
In one embodiment, the angle between the second and third channels and the first channel may be at least one of an acute angle, a right angle, and an obtuse angle.
In one embodiment, the cross section of at least one of the first channel, the second channel, and the third channel may have a rectangular shape having a long axis and a short axis, the long axis may be provided as a horizontal plane, the short axis may be provided as a vertical plane, the long axis may be 10 μm to 10 mm, and the short axis may be 5 μm to 60 μm.
In one embodiment, the fluid containing the cells and the delivery material may have a Reynolds number of 1 to 1,000 according to Equation 1 below in the first channel, and at least one of the second channel and the third channel may have a Reynolds number of 1 to 1,000 according to Equation 1 below.
(In Equation 1, μ is a viscous coefficient of the fluid, ρ is a density, V is an average velocity of the fluid, and D is a hydraulic diameter of the tube.)
In one embodiment, the Reynolds number of the second channel or the third channel may be provided at 40% to 110% of the Reynolds number of the first channel.
In one embodiment, the at least one of the first to third channels may have a hydraulic diameter of 5 μm to 130 μm according to Equation 2 below.
(In Equation 2, Ac is a cross-sectional area of the tube through which the fluid flows, and P is a length of the two-dimensional curve that surrounds and is in contact with the fluid when looking at the cross section.)
In one embodiment, the at least one of the cells and the delivery material of the first channel may have a particle Reynolds number of 4 to 100 defined by Equation 3.
(In Equation 3, Rep is a particle Reynolds number, Re is a Reynolds number, a is a diameter of the cell or particle, and D is a hydraulic diameter.)
In one embodiment, the hydraulic diameter of the second channel or the third channel may be provided at 40% to 110% of the hydraulic diameter of the first channel.
In one embodiment, the at least one of a first vortex and a second vortex may be formed in the vortex region, the first vortex may be formed in a portion where the first channel and the second channel, and the third channel may be connected to each other, and the second vortex may be formed in at least one of the second channel and the third channel.
In one embodiment, the second vortex may be formed in at least one of the second channel and the third channel, and the second vortex may be formed in a portion spaced apart 20 μm to 200 μm in the longitudinal direction of the second or third channel from the central portion of the first channel.
In one embodiment, the at least one of the first vortex and the second vortex may be formed by a change in pressure of the fluid.
In one embodiment, the first vortex or the second vortex may be formed stronger as the Reynolds number of the fluid increases.
In one embodiment, the time during which the flow of the cells is stagnant by the second vortex may be 0.1 μs to 100 μs.
In one embodiment, the at least one of the collision, the first vortex, and the second vortex may form a temporary perforation in at least one of a cell membrane and a nuclear membrane of the cells, and the delivery material may flow into the cells through the perforation.
In one embodiment, the perforation may be formed in at least one of the cell membrane and the nuclear membrane of the cells by at least one of the collision, the first vortex, and the second vortex, the perforation may be formed in the same site or a different site of the cell membrane or the nuclear membrane, a next perforation may be formed after a perforation formed as a first one is maintained or restored, and the perforation formed as a first one may be extended larger or maintained in size by the next perforation.
In one embodiment, the second channel and the third channel may be connected to each other at the end of the first channel to branch the fluid flowing in the first channel, a protruding groove may be formed between the second channel and the third channel, and the protruding groove may be formed by being protruded in a direction in which the fluid flows at a position corresponding to the first channel.
In one embodiment, the protruding groove may have a vertical cross section formed in at least one of a quadrangular shape, a triangular shape, and a cylindrical shape.
In one embodiment, the introduction part of the protruding groove may be 1 μm to 20 μm, and the depth of the protruding groove may be 3 μm to 100 μm.
In one embodiment, the at least one of a first vortex and a second vortex may be formed in the vortex region, the first vortex may be formed by local pressure reversal after the fluid passing through the first channel is collided with at least one of the introduction part of the protruding groove, the inside of the protruding groove, and the partition wall around the protruding groove, and the second vortex may be formed by local pressure reversal in each of the second channel and the third channel.
According to the present disclosure as described above, it is possible to provide a platform for delivering a delivery material into cells by generating multiple vortices. In addition, the delivery material may include various delivery materials such as nucleic acids, proteins, and nanoparticles, and a combination of two or more such as nucleic acids and proteins, in which only one of these is not specified, can provide a platform for intracellular delivery.
According to one aspect of the present disclosure, one embodiment of the present disclosure may include: a first channel through which a fluid containing cells and a delivery material flows while having a flow; and a second channel and a third channel which are connected to the first channel at an angle so that the fluid containing the cells and the delivery material flows while having a flow; wherein the fluid forms at least one of a collision region and a vortex region of the fluid in at least one of the first channel, the second channel, and the third channel.
Details of other embodiments are included in the detailed description and drawings.
Advantages and features of the present disclosure, and methods of achieving them, will become clear with reference to the detailed description of the following embodiments taken in conjunction with the accompanying drawings. However, the present disclosure is not limited to the embodiments disclosed below, but may be implemented in various different forms, and since, unless otherwise specified in the following description, these numbers are essentially approximations that reflect the various uncertainties of measurement that arise in obtaining these values among other things, it should be understood that all numbers, values and/or expressions expressing components, reaction conditions, or amounts of the components in the present disclosure are in all instances modified by the term “about”. Also, when a numerical range is disclosed in the present description, such a range is continuous and includes all values in such a range from a minimum value to the maximum value including a maximum value, unless otherwise indicated. Furthermore, when such a range refers to an integer, all integers including from the minimum value to the maximum value including a maximum value are included unless otherwise indicated.
Further, when a range is stated for a variable in the present disclosure, it will be understood that the variable includes all values within the stated range including the stated endpoints of the range. For example, a range of “5 to 10” includes any subrange of 6 to 10, 7 to 10, 6 to 9, 7 to 9, and the like as well as values of 5, 6, 7, 8, 9, and 10, and it will be understood also to include any value between integers that fall within the scope of the stated range, such as 5.5, 6.5, 7.5, 5.5 to 8.5, 6.5 to 9, and the like. For example, a range of “10% to 30%” includes any subrange of 10% to 15%, 12% to 18%, 20% to 30%, and the like as well as values such as 10%, 11%, 12%, 13% and the like, and all integers including up to 30%, and it will be understood also to include any value between integers that fall within the scope of the stated range, such as 10.5%, 15.5%, 25.5%, and the like.
According to one aspect of the present disclosure, one embodiment of the present disclosure may include: a first channel through which a fluid containing cells and a delivery material flows while having a flow; and a second channel and a third channel which are connected to the first channel at an angle so that the fluid containing the cells and the delivery material flows while having a flow; wherein the fluid forms at least one of a collision region and a vortex region of the fluid in at least one of the first channel, the second channel, and the third channel.
The first channel may be a channel into which a fluid including cells and a delivery material is flowed while having a flow. The fact that the fluid has a flow in the first channel may include flowing it at a constant speed due to an external force being applied along with simply diffusing the fluid. Specifically, the flow of the fluid in the first channel may be controlled by an external force, and thus the cells and the delivery material flowing together with the fluid in the first channel may also flow at a constant speed.
The fluid may represent all fluids flowing through the first to third channels. As the fluid, the first supply fluid and the second supply fluid flow separately in the first channel, but the first supply fluid and the second supply fluid may flow in a mixture in the second or third channel.
A cross section of at least one of the first to third channels may be formed as a rectangle having a short axis and a long axis. The long axis may be provided parallel to the floor with respect to the bottom surface, that is, provided as a horizontal plane. The short axis may be provided almost vertically with respect to the bottom surface, that is, provided as a vertical plane. However, this is only an example, and the shapes of the first to third channels may not be limited thereto.
The material delivery platform may further include: a first supply part supplying a first supply fluid containing at least one of cells and a delivery material; and a second supply part supplying a second supply fluid, wherein the first supply fluid may be supplied to the first channel through a first supply part channel, the second supply fluid may be supplied to the first channel through second supply part channels, one or more of the second supply part channels may be formed, and the second supply part channels may supply the second fluid from both sides of the first supply part channel.
The first supply part may supply the first supply fluid to the first channel. The first supply fluid may be a hydrophilic fluid containing another material. The another material may be at least one of a cell, a delivery material, and a medium.
The first supply part may apply an external force to the first supply fluid to flow the first supply fluid toward the first channel through the first supply part channel. Accordingly, the first supply fluid may flow at a constant speed in the first supply part channel or the first channel.
The second supply part may supply the second supply fluid to the first channel. The second supply fluid may be a hydrophilic fluid, for example, water. The second supply fluid may include a delivery material.
The second supply part may apply an external force to the second supply fluid to flow the second supply fluid toward the first channel through the second supply part channel. Accordingly, the second supply fluid may flow at a constant speed in the second supply part channel or the first channel.
In the first supply part channel or the second supply part channel, the first supply fluid or the second supply fluid may flow while having a certain Reynolds number. The Reynolds number of the first supply part channel and the Reynolds number of the second supply part channel may be 2:1 to 1:3. Preferably, the Reynolds number of the first supply part channel and the Reynolds number of the second supply part channel may be 1:1. When there are two or more second supply part channels, the respective Reynolds numbers of the second supply part channels may be the same.
In the second supply part channel, a plurality of channels in one second supply part may be connected to the first channel. Alternatively, in the second supply part channel, the respective channels in a plurality of second supply parts may be connected to the first channel.
For example, when there is one of the second supply part and there are two second supply part channels connected thereto, the Reynolds number of the second supply part may be twice or more than the Reynolds number of the first supply part. In this case, the Reynolds numbers of the first supply part and the second supply part may be 1:1 to 1:3.
Also, for example, when there are two of the second supply parts and there is one second supply part channel connected to each thereof, the Reynolds number of the second supply part may be the same as or slightly higher than that of the first supply part. In this case, the Reynolds numbers of the first supply part and the second supply part may be 1:1 to 1:1.5.
There may be two or more of the second supply part channels, and the second supply part channels may be formed on both sides with the first supply part channel interposed therebetween. Accordingly, the second supply fluid may be supplied to both sides with the first supply fluid interposed therebetween and supplied to the first channel.
Specifically, in the first channel, the first supply fluid may flow by flowing at the center of the long axis in the first channel, and the second supply fluid may flow by flowing at both sides of the first supply fluid, that is, both ends of the long axis in the first channel. It can flow by floating at the end. For example, when the long axis of the first channel is divided into thirds, the second supply fluid may flow up to a first ⅓ point, the first supply fluid may flow up to a second ½ point, and the second supply fluid may flow by being flowed up to a last ⅓ point again.
Both the first supply fluid and the second supply fluid may include a hydrophilic fluid, but may not be mixed in the first channel. The first supply fluid and the second supply fluid may flow while having a constant speed and a constant Reynolds number, and as a result, the fluids and materials contained in each of the fluids may not be mixed or diffused. In addition, the fact that the first supply fluid and the second supply fluid are not mixed may be influenced by the size and shape of the first channel.
That is, the first supply fluid may be guided to flow in the center of the first channel by the second supply fluid. The first supply fluid may pass through the first channel to collide with a protruding groove formed facing the center of the first channel or near the protruding groove, and thus cells contained in the first supply fluid may collide with the protruding groove or near the protruding groove.
The second channel and the third channel may be channels connected to ends of the first channel.
The second channel and the third channel may be connected to the first channel at a predetermined angle to receive a fluid of the first channel. The angle between the second and third channels and the first channel may be at least one of an acute angle, a right angle, and an obtuse angle. Accordingly, the second channel and the third channel may be formed in an arrow (→) shape, a T shape, or a Y shape based on the first channel. Preferably, the second channel and the third channel may be formed in an arrow (→) shape or a T shape based on the first channel.
The second channel and the third channel may be formed symmetrically or asymmetrically with respect to the first channel. Preferably, the second channel and the third channel may be formed symmetrically with respect to the first channel. In other words, the second channel and the third channel may be formed symmetrically with respect to the longitudinal direction of the first channel.
A cross section of at least one of the first channel, the second channel, and the third channel may have a rectangular shape having a long axis and a short axis, the long axis may be provided as a horizontal plane, the short axis may be provided as a vertical plane, the long axis may be 10 μm to 10 mm, and the short axis may be 5 μm to 60 μm. The length of the short axis or the long axis may be in a range in which the first supply fluid and the second supply fluid can flow without mixing in the first channel. In addition, the length of the short axis or the long axis may be in a range in which vortices can be well formed in the second channel and the third channel.
The first to third channels may be provided at a length ratio of a long axis and a short axis of 2:1, and may be provided in a rectangular shape of, for example, 80×40 μm or 40×30 μm.
The vortex region may be a portion where a straight line-type flow of the first fluid is temporarily stagnant.
The delivery material may be applied without limitation as long as it is a material that can be delivered into cells. The delivery material may be, for example, at least one of nucleic acids, proteins, fluorescent dyes, quantum dots, carbon nanotubes, antigens, ribonucleoproteins, genetic scissors, polymers, and nanoparticles.
The fluid including the cells and the delivery material may have a Reynolds number of 1 to 1,000 according to Equation 1 below in the first to third channels. If the Reynolds number is less than 1, vortexes may not be formed since a sufficient amount and speed are not formed to generate vortices in the channels, and if the Reynolds number is greater than 1,000, an unstable fluid flow may be generated.
In Equation 1, μ is a viscous coefficient of the fluid, ρ is a density, V is an average velocity of the fluid, and D is a hydraulic diameter of the tube.
The Reynolds number of the second channel or the third channel may be 40% to 110% with respect to the Reynolds number of the first channel.
In addition, the second channel or the third channel may be provided to have a Reynolds number smaller than that of the first channel. For example, the second channel or the third channel may be provided to have a Reynolds number that is 50% to 80% of the Reynolds number of the first channel.
In addition, at least one of the first to third channels may have a hydraulic diameter of 1 μm to 100 mm according to Equation 2 below. Also, the hydraulic diameter according to the Equation 2 may be preferably 5 μm to 130 μm. The hydraulic diameter can define the size of each channel. Therefore, when each channel is not a cylinder, the size of each channel can be determined by the hydraulic diameter. The hydraulic diameter in Equation 2 may be the hydraulic diameter in Equation 1 above.
In Equation 2, Ac is a cross-sectional area of the tube through which the fluid flows, and P is a length of the two-dimensional curve that surrounds and is in contact with the fluid when looking at the cross section.
When the channel has a hydraulic diameter of smaller than 5 μm, the amount of fluid flowing in the channel may not be sufficient to form a vortex. In addition, when the channel has a hydraulic diameter of larger than 130 μm, a force (for example, pressure reversal) capable of forming a vortex may not be formed in the flow of the fluid.
The second channel or the third channel may be provided to have a hydraulic diameter that is 40% to 110% based on the hydraulic diameter of the first channel.
In addition, the second channel or the third channel may be provided to have a hydraulic diameter of smaller than that of the first channel. For example, the second channel or the third channel may be provided to have a hydraulic diameter of that is 50% to 80% of the hydraulic diameter of the first channel.
In addition, the fluid of the first channel may be defined by a particle Reynolds number (Rep) of Equation 3.
(In Equation 3, Rep is a particle Reynolds number, Re is a Reynolds number, a is a diameter of the cell or particle, and D is a hydraulic diameter.)
For example, when the particle Reynolds number is about 1, the first supply fluid delivered to the first channel may flow by flowing in the center of the first channel. Accordingly, in this case, the cells may flow to the center of the first channel even without the second supply part channel.
In addition, for example, when the particle Reynolds number is between 4 and 100, the second supply fluid guides the first supply fluid so that the first supply fluid may flow by flowing in the center of the first channel.
Accordingly, the particle Reynolds number in the first channel may be 0.5 to 100. In addition, the particle Reynolds number in the first channel may be 1 to 100. In addition, the particle Reynolds number in the first channel may be preferably 4 to 100. In addition, the particle Reynolds number in the first channel may be preferably 25 to 40.
At least one of portions where the first to third channels and the first channel and the second and third channels meet may include a vortex region. The straight line-type flow of the fluid flowing through the first to third channels may be temporarily stagnant in the vortex region. The vortex region may be a portion where a vortex is formed in the channel.
A protruding groove may be formed in a portion where the first channel is connected to the second and third channels. The protruding groove may be formed by protruding in a direction in which a fluid flows at a position corresponding to the first channel.
The protruding groove may diverge a fluid flowing in the first channel by connecting the second channel and the third channel to each other at an end of the first channel.
The protruding groove may have a vertical cross section formed of at least one of a quadrangular shape, a triangular shape, and a cylindrical shape. The protruding groove may be in the form of a tube with a closed end, and may not be limited to the above example as a form capable of generating a vortex.
In addition, the introduction part of the protruding groove may be 1 μm to 20 μm, and the depth of the protruding groove may be 3 μm to 100 μm.
The cells coming out of the first channel may collide with the introduction part of the protruding groove or may be flowed into the protruding groove and collide with each other. Even when the cells flow into the protruding groove and collide with each other, they may escape out of the protruding groove after the collision and undergo a vortex.
Alternatively, the cells may collide with a partition wall around the protruding groove.
At least one of a first vortex and a second vortex may be formed in the vortex region. Specifically, the first vortex may be formed in a portion where the first channel and the second channel are connected and a portion where the first channel and the third channel are connected to each other. Also, specifically, the second vortex may be formed in at least one of the second channel and the third channel.
The vortex may refer to a flow in which the fluid encounters a sudden change in pressure, an obstacle, etc. to interrupt the flow so that the proceeding direction is changed. That is, the vortex may be formed while the fluid flowing through the channel collides with an obstacle (e.g., a protruding groove or a partition wall) or a straight line flow is stagnant due to reversal of pressure, etc. Accordingly, when a vortex is formed in the fluid, cells and/or a delivery material contained in the fluid may receive an external force while undergoing the vortex, and in this process, deformation may occur in the cells.
In more detail, the first vortex may be formed in a stagnation point region. The stagnation point region may be included in a portion where the first channel is connected to the second and third channels. The stagnation point region may be a portion where the fluid flowing out of the first channel is stagnant to form the first vortex. That is, the stagnation point region may be a portion where the fluid flowing out of the first channel collides with at least one of the protruding groove and the partition wall to form a first vortex. Accordingly, the stagnation point region may be a portion where the cells and/or the delivery material contained in the fluid collide with at least one of the protruding groove and the partition wall and undergo a first vortex. However, the cells may undergo only one of the collision and the first vortex, and even in this case, deformation may occur in the cells.
“Collision” of the cells may be a meaning including all that the cells collide with at least one of the introduction part of the protruding groove, the inside of the protruding groove, and the partition wall. Here, the partition wall may be a partition wall around the protruding groove.
In addition, the second vortex may be formed in at least one of the second channel and the third channel. A site where the second vortex is formed may be a site completely out of the end of the first channel. In other words, the site where the second vortex is formed may be a site completely out of a region extending in the same width as the width of the first channel in the longitudinal direction of the first channel. Specifically, the site where the second vortex is formed may be formed in a portion spaced apart from the central portion of the first channel (or the central portion of the protruding groove) by 20 μm to 200 μm in the longitudinal direction of the second or third channel.
At a point ahead of 20 μm in the longitudinal direction of the second channel (or the third channel), the first fluid flowing in the first channel may interfere with the formation of a vortex so that the vortex may not be formed well. In addition, at a point behind than 200 μm in the longitudinal direction of the second channel (or the third channel), the fluid may not have a sufficient velocity or Reynolds number to form the vortex.
The second vortex may be formed by a change in pressure of the fluid in the second channel or the third channel, and the flow of the fluid may be stagnant at a portion where the second vortex is formed. Therefore, the flow of the cells and the delivery material contained in the fluid may be stagnant.
The fact that the flow of the fluid, cells, and delivery material is stagnant (or stopped) may mean that the straight line flow is stagnant, and may not mean a state without movement. When the flow of the fluid, cells and delivery material is stagnant, they can move in a vortex shape riding on the flow of the vortex. That is, the fluid, cells, and delivery material may be stagnant by the vortex.
The time at which the cells are stopped (stagnant) by the second vortex may be 0.1 μs to 100 μs.
The first vortex or the second vortex may be formed stronger as the Reynolds number of the fluid increases. Accordingly, when the Reynolds number of the first channel is greater than that of the second channel and/or the third channel, the first vortex may be formed to be stronger than the second vortex. In this case, the cells may be deformed more by the first vortex.
The fluid flowing through the first to third channels and the cells and delivery material contained in the fluid may undergo at least one of the collision, the first vortex, and the second vortex. Therefore, even when the cells undergo only one of the collision, the first vortex, and the second vortex, the cells may be deformed and the delivery material may be delivered into the cells.
When cells are deformed by the collision or vortex, a temporary perforation may be formed in at least one of a cell membrane and a nuclear membrane, and the delivery material may flow into the cells through the perforation.
At this time, the perforation may be formed in the same site or different site of the cell membrane or the nuclear membrane, and a next perforation may be formed after a perforation formed as a first one is maintained or restored. The perforation formed as a first one may be extended larger by the next perforation.
The cells may undergo all of the collision, the first vortex, and the second vortex, and at this time, the delivery material may be better delivered into the cells.
When the delivery material is delivered into the cells by the collision or vortex, the delivery material may be delivered into the cells when deformation occurs only in the cells. However, when deformation occurs in the cell membrane and the nuclear membrane, the delivery material may be delivered to at least one of the inside of the cells and the inside of the nucleus.
The delivery material may include all materials having a size smaller than that of the cells. The delivery material may be, for example, at least one of nucleic acids, proteins, and nanoparticles. Specifically, the delivery material may be each composed of nucleic acids, proteins, and nanoparticles, and may include various combinations of one or more, such as a combination of nucleic acids, proteins, and nanoparticles, depending on the use.
At least one of a first vortex and a second vortex may be formed in the vortex region, the first vortex may be generated after the fluid passing through the first channel collides with the protruding groove or the partition wall, and the second vortex may be formed by a local increase in pressure in each of the second and third channels.
Specifically, the cells may receive a two-step force in the stagnation point region. As a first step, the cells collide with the introduction part of the protruding groove or the inside of the protruding groove, or collide with the partition wall to receive a force, and thus deformation may occur in the cells. As a second step, the cells leave the protruding groove or the partition wall, and the flow of the fluid leaving the protruding groove or the partition wall is mixed with the flow of the first channel to form a first vortex, and thus deformation may occur in the cells. However, the cells may undergo only one of the first and second steps.
Since the cells undergo the first vortex after collision, they may be deformed to a greater extent in the stagnation point region. In addition, the force by which the first vortex deforms the cells may be greater than the force by the second vortex.
Meanwhile, the second vortex may be generated when a pressure increases in a local portion of the second channel and/or the third channel. This is because the fluid flows from a high pressure side to a low pressure side in the second channel and/or the third channel, but there may be a portion where the pressure is reversed in a local portion of the second channel (or third channel), where the second vortex may be formed.
The first vortex and the second vortex may be sequentially formed based on one of the cells or delivery material. However, the first vortex and the second vortex may be formed simultaneously or at different times based on the platform according to one embodiment of the present disclosure.
Hereinafter, examples and comparative examples of the present disclosure will be described. However, the following examples are only preferred embodiments of the present disclosure, and the scope of rights of the present disclosure is not limited by the following examples.
A plurality of second supply part channels in one second supply part may be connected to the first channel (upper
Both of the first supply fluid and the second supply fluid may include fluids of the same nature, but the fluids are not mixed or the material exchange is not performed due to the velocity or channel shape of these fluids.
In addition,
In
mRNA expressing green fluorescent protein (GFP) was delivered to K562 cells. In
In addition, the results in
Those skilled in the art to which the present disclosure pertains will understand that the present disclosure can be embodied in other specific forms without changing its technical spirit or essential features. Therefore, the embodiments described above should be understood as illustrative in all respects and not limiting. The scope of the present disclosure is indicated by the claims to be described later rather than the detailed description above, and all changes or modified forms derived from the meaning and scope of the claims and equivalent concepts thereof should be interpreted to be included in the scope of the present disclosure.
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2021-0047258 | Apr 2021 | KR | national |
| 10-2022-0044922 | Apr 2022 | KR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/KR2022/005272 | 4/12/2022 | WO |
