KR101347134B1 - Device of Directional Property Localization and Control of Magnetic Field for Differentiation and/or Proliferation of Stem Cell - Google Patents

Abstract

The present invention is a method and apparatus for moving and immobilizing stem cells to an injured site using magnetism, or injecting and immobilizing stem cells directly to the injured site, and differentiating or proliferating the fixed stem cells into neurons. It is about. By using the present technology, it is possible to differentiate or proliferate stem cells into neurons by applying an appropriate magnetic field to a site where nerves are damaged, and to control the growth direction of the cells. The device proposed in the present invention is composed of multi-channels, and controls the stimulation site by moving the bed or adjusting the multi-channel magnetic field formation region. In addition, the alternating method of the magnetic field (direct current, DC) and the weak magnetic field (AC, AC) allows direct current and alternating current to flow at the same time or only direct current or alternating current, and thus directs (direct action) and differentiation or proliferation of stem cell differentiation and proliferation. Adjust the (AC) action. Through the technique of the present invention, it is expected to contribute to increase the efficiency of differentiation or proliferation of stem cells into nerve cells at the site of injury.

Description

Device of Directional Property Localization and Control of Magnetic Field for Differentiation and / or Proliferation of Stem Cell}

The present invention is a method and apparatus for moving and immobilizing stem cells to an injured site using magnetism, or injecting and immobilizing stem cells directly to the injured site, and differentiating or proliferating the fixed stem cells into neurons. It is about. By using the present technology, it is possible to differentiate or proliferate stem cells into neurons by applying an appropriate magnetic field to a site where nerves are damaged, and to control the growth direction of the cells. The device proposed in the present invention is composed of multi-channels, and controls the stimulation site by moving the bed or adjusting the multi-channel magnetic field formation region. In addition, the alternating method of the magnetic field (direct current, DC) and the weak magnetic field (AC, AC) allows direct current and alternating current to flow at the same time or only direct current or alternating current, and thus directs (direct action) and differentiation or proliferation of stem cell differentiation and proliferation. Adjust the (AC) action. Through the technique of the present invention, it is expected to contribute to increase the efficiency of differentiation or proliferation of stem cells into nerve cells at the site of injury.

The present invention relates to a method and apparatus for moving stem cells by using magnetism, fixing them to a desired site, and then differentiating or proliferating them. Instead of moving the stem cells to the injury site, a method of directly injecting the stem cells to the injury site is also included.

The device proposed in the present invention is composed of a multi-channel, and adjusts the stimulation site by moving the bed or by adjusting the multi-channel magnetic field forming region, and forms a magnetic field in the alternating manner of the magnetic field and the weak magnetic field, the ferromagnetic field Is DC (direct current), and the weak magnetic field is formed through AC (AC). In addition, the magnetic field is characterized in that it is formed through a permanent magnet, an electromagnet, the Helmholtz method. When forming a magnetic field with an electromagnet, two coils are wound around the same trajectory, one flowing a DC current, and the other flowing an AC current, which sends both the DC current and the AC current together. In this case, DC is used to control stem cell migration and the direction of differentiation and proliferation, and AC is used to promote differentiation or proliferation of stem cells. By controlling the size and position of the coil and the current flowing through the coil, it is possible to change not only the strength of the magnetic field but also the distribution characteristics, so that local magnetic field generation is possible.

Stem cell refers to the undifferentiated cells of the stage prior to differentiation into each cell constituting the tissue, can divide and self-renewal for a long time, and various kinds given conditions It is a cell that can differentiate into cells. Stem cells are largely divided into embryonic stem cells and adult stem cells according to the origin of the tissue. Embryonic stem cells are stem cells derived from human embryos and have the ability to differentiate into almost all kinds of cells (pluripotent). Cells that can develop into a variety of cells and tissues derived from ectoderm, mesoderm, and endodermal layers, which originate from the inner cell mass located inside the blastocyst appearing after 4-5 days of fertilization, and into various other tissue cells. Differentiate but do not form new life.

Adult stem cells have a multipotent ability to differentiate into cells of a limited variety of cells derived from each organ of the adult, and can only differentiate into cells specific to the tissues and organs that contain them. As stem cells, they are involved in the growth and development of individual tissues and organs in prenatal, neonatal and adulthood as well as in maintaining the homeostasis of adult tissues and inducing regeneration in tissue damage and collectively refer to tissue-specific pluripotent cells. Adult stem cells have limited disadvantages than embryonic stem cells but have no ethical problems and no side effects, and many therapeutic agents have been studied for adult stem cells.

An electromagnet is a magnet that magnetizes when electricity flows and returns to its original, unmagnetized state when the current is interrupted, which always maintains magnetism regardless of the supply of current. It is distinguished from permanent magnets. When a current flows through the wire, a concentric magnetic field is formed around the wire. Using this principle, a very strong magnetic field cannot be obtained with a permanent magnet. The electromagnet can change the strength of the magnetic field relatively easily by artificially adjusting the current. A magnetic field is a space in which magnetic force acts around a magnet, an electric current, a changing electric field, etc., and is also called a magnetic field or a magnetic field.

There are prior arts related to the transfer and fixation of stem cells using magnetism and the differentiation or proliferation of stem cells (10-2008-0002096, 10-2009-011101, 10-2009-0110102). A technique for observing stem cell suspension on a plate from the outside and observing the control effect on the movement, fixation, differentiation and proliferation of stem cells by magnetism, or injecting the stem cells into a microchannel before using them in a living body and using an external magnetic field The present invention relates to a simulation apparatus capable of observing the control effect on the movement, fixation, differentiation and proliferation of stem cells, wherein stem cells are introduced directly into a living body, and the movement, fixation, and differentiation of stem cells in a living body through a magnetic field. And techniques for available devices that can control proliferation are absent.

In addition, there are existing techniques (US20060205993, JP 2008-543388, US200740065941, US20050075679) that use magnetism for neurotherapy, but these techniques are performed by directly implanting electrodes into a living body. It is accompanied by pain.

Therefore, it can be utilized in the human body, and it can form a magnetic field outside the body and form a magnetic field locally at the site to be stimulated to move and fix the stem cells to the damage site, and to differentiate or proliferate into nerve cells. Skill is needed.

Thus, in the present invention, as a device that can be utilized in the human body, it is possible to form a magnet outside the living body and apply a magnetic field locally to a site to be stimulated to move and fix the stem cells to the site of injury, to differentiate or proliferate. It is intended to provide a description of the method and apparatus.

In order to achieve the above technical problem, the present invention uses magnetism outside the body to locally apply a magnetic field to a site to be stimulated, thereby moving stem cells to an injury site, or injecting stem cells directly into an injury site. The present invention provides a method and apparatus for immobilization, differentiation or propagation.

According to another aspect of the invention, the first step of injecting stem cells into the damaged area, and the radial interval of the channel, the coil to be turned on / off according to the target of the injection of the stem cells, and the current to flow in the channel according to the exposure intensity A second step of adjusting the stimulation site by adjusting the height and the left and right of the bed according to determining the amount of the local exposure site of the magnetic field, and a third step of flowing a current to the coils around the stimulation site, Directional localization and directional control method of the magnetic field for stem cell differentiation or proliferation, characterized in that it comprises a fourth step of moving and fixing the stem cells to a desired position using the magnetic field formed by the current, differentiation or proliferation Provide technology on

Still another aspect of the present invention provides a method for directing localization and direction control of a magnetic field for stem cell differentiation or proliferation, characterized in that in the first step, stem cells are injected or directly injected into a blood vessel connected to an injury site. Provide the technology.

According to a third aspect of the present invention, the stem, characterized in that the movement, fixation, differentiation and proliferation of the stem cells in the fourth step is controlled by a magnetic field formed when the direct current and the alternating current are flowed at the same time. The present invention provides a technique for directional localization and direction control of a magnetic field for cell differentiation and proliferation.

Fourth configuration of the invention, the injury site is Alzheimer's disease, depression, Parkinson's disease, cerebral infarction, cerebral hemorrhage, nerve or spinal cord injury site, characterized in that the directional localization and direction of the magnetic field for stem cell differentiation and proliferation Provides a technique regarding a control method.

The fifth configuration of the invention, the bed in which the stem cell injection target is located, a circular or semi-circular cylinder located on the lower surface of the bed, the two coils are wound in the same trajectory in a pair of coils, the 2 A coil wound in pairs of strings, each of which is characterized by simultaneously flowing a direct current and an alternating current (AC), the current unit and the position of the bed and the multi-channel current, characterized in that the channel is configured as one unit, the multi-channel It provides a device for directional localization and directional control of the magnetic field for stem cell differentiation, proliferation, characterized in that it comprises a control unit for controlling the.

In a sixth aspect of the present invention, there is provided a device for directional localization and directional control of a magnetic field for stem cell differentiation and proliferation, wherein the radius of the channel and the distance between the multichannels are changed according to the desired intensity and distribution characteristics of the magnetic field. to provide.

According to a seventh aspect of the present invention, the directionality of the magnetic field for stem cell differentiation and proliferation is characterized in that the current flowing through the multichannel is turned on / off in sequence or alternately according to the desired intensity and distribution characteristics of the magnetic field. Provides a localization and directional control device.

Hereinafter, with reference to the drawings, embodiments, and the like with respect to the problem solving means of the present invention will be described in detail.

As described above, the present invention provides a method and apparatus for ex vivo to use magnetic to apply a magnetic field locally to a site to be stimulated to move stem cells to a site of injury and to differentiate or proliferate into neurons. It is expected that the present invention can contribute to the effective differentiation or proliferation of stem cells into neurons without implanting an electrode in a target for neural cell differentiation or proliferation.

1 is a diagram illustrating an apparatus for differentiating or proliferating stem cells that can be utilized through a method of moving and immobilizing stem cells using magnetism and differentiating or proliferating into neurons.
2 is a diagram in which a coil, which is an alternating system of direct current and alternating current, is wound around a cylinder.
Fig. 3 shows that the coil wound cylinder is one channel, which is a unit, and is composed of multiple channels, and shows a distribution of the magnetic field formed around the coil at this time.
4 is a schematic diagram showing the density and distribution characteristics of the magnetic field formed according to the radius of the channel and the distance between the channels.
FIG. 5 is a simulation diagram showing the distribution of a magnetic field formed according to ON / OFF of a channel or according to an interval between channels.
6 is a simulation diagram showing the distribution of the magnetic field formed according to the radius of the channel.
7 is a differentiation photograph of bone marrow-derived mesenchymal stem cells cultured by exposure to a magnetic field.
In FIG. 8, FIG. 8A is a schematic diagram showing that the stem cells differentiate into nerves through the steps of neural stem cells, and FIG. 8B is a gene analysis diagram confirming the expression of Nestin, which is a marker of neural stem cells.

In order to carry out the present invention, first, a method of transferring and immobilizing stem cells by using magnetism, differentiating or proliferating them into nerve cells, and suggesting a method of differentiating or proliferating nerve cells at a nerve injury site is proposed. Differentiation and propagation method consists of 6 steps.

<How to move and fix stem cells using magnetism and differentiate or proliferate into neurons>

1. Step 1: Inject stem cells into the damaged area.

① At this time, the damage site, Alzheimer's disease, depression, Parkinson's disease, cerebral infarction, cerebral hemorrhage, or nerve injury or spinal cord injury is characterized in that the site.

② Stem cells are injected into the blood vessels connected to the damaged area or directly injected into the damaged area.

2. Step 2: Depending on the stem cell injection subject, determine or adjust the following.

① Determine the radius of the channel to be used, the interval, and the channel to be turned on or off, depending on the stem cell injection target.

② Determine the amount of current to flow in the channel according to the exposure intensity, depending on the subject injected the stem cells.

③ Determine the local exposure site of the magnetic field according to the target of the stem cell injection.

④ Adjust the stimulation site by adjusting the height and left and right of the bed, according to the local exposure site of the magnetic field (Fig. 1).

3. Step 3: The current flows through the coil around the magnetic pole area.

① The coil is characterized in that the coil is wound in a circular or semi-circular cylinder, and if the magnetic pole is to be uniformly magnetized, the circular is selected, and if the magnetic pole is to be strongly strongly selected, the semi-circle is selected.

(2) The coil wound around the cylinder is characterized by being wound in pairs of two rows with the same trajectory (Fig. 2).

(3) The coil wound in pairs of the above two lines is characterized in that direct current and alternating current are flowed simultaneously or only direct current or alternating current (Fig. 2).

④ The coil is wound into a single channel, and the channel is configured as a unit, multi-channel to adjust the magnetic field (Fig. 3).

(5) The radius of the channel (FIGS. 4 and 6) and the distance between the channels (FIGS. 4 and 5) are changed according to the intensity and distribution characteristics of the desired magnetic field.

(6) The current flowing in the multi-channel is characterized in that the ON / OFF, in turn or alternately the channels according to the desired strength and distribution characteristics of the magnetic field (Fig. 5).

4. Step 4: Using the magnetic field generated by the electric current, the stem cells are moved and fixed to the damaged area, differentiated or expanded.

In the fourth step, the movement, fixation, differentiation and proliferation of stem cells are characterized in that they are controlled by a magnetic field formed when direct current and alternating current are flowed at the same time.

Example 1 Differentiation of Stem Cells Using Magnetics

To investigate the effect of the magnetic field on the differentiation or proliferation of stem cells, bone marrow-derived mesenchymal stem cells were cultured by exposing them to magnetic fields.

① The bone marrow-derived mesenchymal stem cells were passaged for 12 days to observe their morphology.

② In the subculture, low glucose DMEM was used as a minimal medium, and EGM media was used as a differentiation medium. The EGM media is characterized in that the endothelial media contains a variety of cytokine and growth factors, the EGM media is a medium capable of differentiating mesenchymal stem cells into other cells.

③ Bone marrow-derived mesenchymal stem cells cultured in the medium were continuously exposed to electromagnetic waves for 12 days.

④ As a result of the exposure of the electromagnetic wave, it was observed that filament similar to neurite was seen in mesenchymal stem cells (FIG. 7).

⑤ The mesenchymal stem cells are characterized in that they are differentiated into nerves through neural stem cells (step 8A) before being differentiated into neurons. Therefore, expression of Nestin, a marker of neural stem cells, was confirmed. At this time, a comparative analysis was performed using Neuroblastoma cell line, a neurocancer cell line, as a positive marker. The results for this comparative analysis are shown in Figure 8B.

Next is the configuration of a device for treating nerve damage by moving, fixing, differentiating and proliferating stem cells using magnetism.

<Device to move and fix stem cells using magnetism and to differentiate or proliferate into neurons>

The nerve injury treatment apparatus is characterized in that it is largely composed of a bed, a current unit, a control unit as follows.

1. Bed

Bed where the stem cell injection target is located, its position can be adjusted in the control unit. In other words, the bed moves up or down depending on the part to be stimulated so that the magnetic field can be locally applied only to the part to differentiate or proliferate the stem cells.

2. Current section

The bottom of the bed is located,

The coil is wound in the same trajectory with two pairs of coils in a pair, and the coils wound in the two pairs of pairs of direct current and alternating current are simultaneously flown.

The current unit is characterized in that the channel is configured in one unit, multi-channel.

The radius of the channel and the distance between the multiple channels can be changed according to the intensity and distribution characteristics of the desired magnetic field. The strength and distribution of the magnetic field formed according to the change of the radius of the channel can be confirmed through the simulation data of FIG. 6, and the density and distribution of the magnetic field according to the change of the interval between the multi-channels can be checked through FIG. 5.

The change in the magnetic field due to the change in the distance between the channels will be described in Example 3, and the change in the magnetic field due to the change in the radius of the channel will be described in more detail in the fourth embodiment.

3. Control

The control unit adjusts the position of the bed and the current of the multi-channel.

The position of the bed, by moving the bed in the up or down direction according to the area to be stimulated so that the magnetic field can be applied locally only to the area to differentiate or proliferate stem cells.

The current adjustment of the multi-channel is turned on / off in sequence or alternately according to the desired intensity and distribution characteristics of the magnetic field. The intensity and distribution of the magnetic field formed by the ON / OFF method of the multi-channel current can be confirmed through the simulation data of FIG. 5.

The change in the magnetic field through the current ON / OFF method of the channel will be described in more detail with reference to Example 2.

<Control unit of a device that moves and fixes stem cells using magnetism and differentiates or proliferates into nerve cells>

1. Power source of control unit

The power source refers to a device capable of arbitrary waveform formation (frequency, magnitude, shape, etc.) up to a large current and a high voltage, and controls the intensity of a current flowing through a coil by using the power source.

2. Solid state relay of control unit

The solid state relay refers to a device that controls ON / OFF of high voltage and high current through the low voltage output of the microcontroller. The solid state relay controls ON / OFF of the current flowing through the coil using the solid state relay.

In the method and apparatus for moving and immobilizing stem cells by using the magnetism and differentiating or proliferating into neurons, the current of a channel is turned on or off in turn or alternately, or the radius of the channel and the distance between the channels are changed. Changes in the strength and distribution characteristics of the magnetic field according to the following Examples 2 to 4 will be described in more detail.

Example 2 magnetic field change through the current ON / OFF method of the channel

By turning ON / OFF the current of five channels, the strength and distribution characteristics of the magnetic field can be changed. At this time, the number of channels is not limited to five, and the number can be changed.

1) In a large area, if a strong stimulus as a whole is desired, turn on the currents of all five channels (turn on the channels 1 to 5). The strength and distribution of the magnetic field formed at this time can be confirmed through the simulation result of FIG. 5A.

2) In a large area, if weak stimulation as a whole is desired, five channels are alternately turned on / off (channels 1, 3, and 5 are turned on, and channels 2 and 4 are turned off). The strength and distribution of the magnetic field formed at this time can be confirmed through the simulation result of FIG. 5B.

3) In a narrow area, if stimulation is desired locally, two consecutive channels are turned on in turn, and the remaining channels are turned off. The strength and distribution of the magnetic field formed at this time can be confirmed through the simulation result of FIG. 5C.

Example 3 Magnetic Field Variation by Changing the Distance Between Channels

According to the ON / OFF method, it is possible to adjust the spacing between the channels, through which the density and distribution characteristics of the magnetic field can be changed.

1) If the spacing between the channels is narrowed, the magnetic field density is high and the distribution area can be narrowed as compared with the case where the spacing between the channels is relatively large (FIG. 4A).

2) When the spacing between the channels is wider, the density of the magnetic field is lower and the distribution area can be wider than when the spacing between the channels is relatively narrow (FIG. 4B). The density and distribution characteristics of the magnetic field formed according to the distance between the channels can be confirmed through the simulation data of FIG. 5.

Example 4 Change of the Magnetic Field by Changing the Radius of the Channel

By changing the radius of the channel, it is possible to change the density and distribution characteristics of the magnetic field.

1) If the radius of the channel is shortened, the magnetic field density is higher and the distribution area is narrower than that of the relatively long radius of the channel (FIG. 4A). The density and distribution region of the magnetic field formed at this time can be confirmed through simulation data of FIG. 6B.

2) When the radius of the channel is increased, the magnetic field has a lower density and a wider distribution area than the case where the radius of the channel is relatively short (FIG. 4B). The density and distribution region of the magnetic field formed at this time can be confirmed through the schematic diagram of FIG. 6A.

Claims (7)

delete delete delete delete Bed in which the subject to inject stem cells is located;
A circular or semi-circular barrel positioned to surround the bottom of the bed;
A channel configured to wind two coils in a pair with the same trajectory, and flow a direct current to one coil and an alternating current to another coil to form a magnetic field;
A current unit comprising the channel as one unit and configured as a multi-channel; And
Directional localization and direction control device of the magnetic field for stem cell differentiation or proliferation comprising a control unit for controlling the position of the bed and the current of the multi-channel. 6. The method of claim 5,
Directional localization and direction control device of the magnetic field for stem cell differentiation or proliferation, characterized in that for changing the radius of the channel and the interval between the multi-channel, depending on the intensity and distribution characteristics of the desired magnetic field. The method according to claim 5 or 6,
Directional localization and direction control device of the magnetic field for stem cell differentiation or proliferation, characterized in that the current flowing through the multi-channel is turned on / off, depending on the intensity and distribution characteristics of the magnetic field, in turn or alternately.

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