KR20080009911A - Syringe for cell injection - Google Patents

Abstract

A syringe for cell injection is provided to transplant a minute amount of cells into the cell, tissue or internal organ of an animal effectively and minimize the damage of the tissue. A syringe for cell injection includes a cylinder(10), a piston(20) and a discharge hole(30). The discharge hole of the syringe is connected to a capillary tube(50). The capillary tube is made of glass. The discharge hole and the capillary tube are fixed with an adhesive and/or a curing agent. Cells are included inside the cylinder of the syringe. The cells are stem cells. The content of the cells contained inside the cylinder of the syringe is in the range of 1-5 micro-liters. When a user pushes a pushing rod(40), the piston connected to the pushing rod advances along the inner wall of the cylinder to inject injection material.

Description

Syringe for Cell Injection

1 is a front view showing a cell injection syringe of the present invention.

Figure 2 is a diagram observing the transplantation of stem cells into the mouse brain using the cell injection syringe of the present invention. Specifically, <A> is a fluorescence micrograph of a brain tissue transplanted with stem cells, and <B> is a fluorescence micrograph of a brain tissue without transplanted stem cells.

<Explanation of symbols for the main parts of the drawings>

10: syringe cylinder

20: piston

30: discharge port

40: push

50: capillary

The present invention relates to a cell injection syringe, and more particularly to a cell injection syringe having a capillary connected or attached to the discharge port. Cells sucked into the syringe of the cell injection syringe of the present invention are injected into the desired cells, tissues, organs or organs through a fine capillary connected to the needle of the syringe.

In particular, the present invention can be usefully used to confirm the differentiation or therapeutic ability of stem cells by injecting stem cells into specific cells, tissues or organs.

In the medical field, many developments have been made, but there are diseases that cannot be cured by modern medicine, that is, incurable diseases. Patients with these intractable diseases often lose their function, partially or entirely, by tissues and cells that should function normally. In this case, it is true that fundamental treatment is impossible using any known drug or surgical treatment.

Previously, when organ function was partially paralyzed or completely lost, the last option was to transplant the organ. In this case, it is difficult to find an organ suitable for the patient, and there is a problem that a lifelong immunosuppressant should be used due to the immune rejection reaction. Immunosuppressants not only reduce the quality of life of the patient, but also cause a decrease in immunity, which eventually leads to various complications. However, as the researches related to gene transplantation, cell transplantation, and stem cell transplantation have been actively conducted, the parts which have been pointed out as problems in the existing treatment methods have been improved. These studies are notable for the purpose of cosmetic treatment as well as for the treatment of intractable disease. This is achieved. An example is the treatment of intractable disease through stem cell transplantation. Because stem cells are identical to the patient's own genes, they can solve not only the treatment of the disease but also the immune rejection reactions of existing organ transplants. Another example is micro regenerative cell injection, which is being carried out for cosmetic or therapeutic purposes to achieve cosmetic effects. Instead of using conventional adipose tissue transplantation and filler implants, it is possible to fundamentally solve the wrinkles of the skin by selecting and transplanting only the fat cells treated in the adipose tissue, thereby recovering healthy skin with a simple injection without side effects.

Cell injection or transplantation is a breakthrough and important technology in that it enables the treatment of intractable disease. And when it is backed by delicate and accurate techniques and instruments, it will be possible to reach the common goal of humankind in the treatment of incurable diseases.

Conventionally, syringe needles have been used directly when injecting cells in vivo. This method can greatly damage tissues of animals, and in some cases, a hole caused by the injection needle may exist, and the injected cells may flow out again. there was.

In order to overcome this problem, the inventor of the present invention has developed a syringe for accurate and convenient cell injection in the process of studying how to transplant stem cells to effectively inject or transplant a small amount of cells into the cells, tissues, organs or organs of the animal. The present invention has been completed by finding a method.

Accordingly, the present invention provides a syringe for cell injection, and the present invention also provides a method for effectively implanting a small amount of cells into cells, tissues, organs or organs of the animal.

In order to achieve the above object, the present invention provides a syringe for cell injection. That is, the cell injection syringe of the present invention is a syringe including a piston and an outlet, characterized in that the discharge port of the syringe is connected by a capillary tube. In addition, the cell injection syringe of the present invention may include cells in the cylinder of the syringe.

More specifically, the present invention is a syringe configured to inject the injection material through the discharge port 30 while the piston 20 connected to the push rod advances along the inner wall of the syringe cylinder 10 by pushing the push rod 40, It provides a syringe for cell injection, characterized in that the discharge port of the syringe is connected by a capillary.

The capillary is characterized in that the material is glass, the cell comprises a stem cell.

It also includes the outlet and the capillary is fixedly connected with an adhesive and / or hardener.

The content of cells to be injected into the syringe includes those in the range of 1 to 5 μl.

It includes including the cells in the cylinder of the syringe, the content of the cells contained in the cylinder of the syringe is characterized in that in the range of 1 ~ 5 μl.

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

Figure 1 shows a front view of the syringe for cell injection of the present invention. In the syringe for cell injection according to the present invention, in the syringe configured to inject the injection material through the outlet 30 while the piston 20 connected to the push rod is pushed along the inner wall of the syringe cylinder 10 by pushing the push rod 40, Capillary tube 50 is connected to or attached to the discharge port 30. Capillary tube 50 can be selected from a variety of materials, glass (glass) is preferred.

The discharge port 30 and the capillary tube 50 of the syringe for cell injection according to the present invention select the capillary tube 50 having an outer diameter slightly larger than the outer diameter of the discharge port 30 so that the outer diameter of the discharge port and the inner diameter of the capillary tube closely adhere to each other. Can be connected as possible. In addition, adhesives and / or hardeners may be used to firmly bind the outlet and capillary and prevent leakage of cells in the syringe. The adhesive includes an adhesive commonly used in the art such as an animal adhesive, a vegetable adhesive, a synthetic resin adhesive, a rubber adhesive, a fibrous adhesive, a hot melt adhesive, an epoxy, an acrylic adhesive, and the like. In particular, in the case of the present invention, Araldite rapid (Araldite rapid) type is preferred, it is most preferable to use a mixture of the Araldite rapid and the curing agent.

On the other hand, an example of the use aspect of the cell injection syringe of this invention is demonstrated as follows. That is, a method of injecting stem cells into the brain of a mouse using the cell injection syringe of the present invention will be described.

(a) a first step of inhaling stem cells into the syringe of the present invention; (b) a second step of fitting the glass capillary into the syringe outlet; And (c) a stem cell transplantation method comprising the third step of transplanting stem cells into a disease model animal.

The type of cell and the animal to which the cell injection syringe of the present invention can be used are not limited. In addition, the cells to which the present invention can be applied include all cells constituting the living body, that is, somatic cells and germ cells, and preferably include stem cells. In addition, animals to which the syringe for cell injection of the present invention may be applied include mammals including humans, reptiles, fish, amphibians and birds, and are preferably mammals. In addition, tissues, organs or organs in which the syringe of the present invention may be used include liver, stomach, spleen, pancreas, small intestine, large intestine, duodenum, kidney, uterus, heart, lung, brain, eye, skin, muscle, nerve and blood vessel It includes.

Hereinafter, a preferred embodiment of the present invention will be described in more detail.

Example  1. Preparation of Animals

^{Colonies of} BALB / c npc ^nih (NP-C) mice were maintained and genotypes were confirmed by PCR using genomic DNA isolated from small tail tails as described in Loftus et al., 1997. It was.

Example  2. Obtaining Stem Cells

To obtain meschymal stem cells (Ms-MSCs) of the mice, the tibia and femur of 6-8 week-old heterozygous Phgdh-DsRed2 male mice were dissected. Bone marrow cells were collected and stored in mesodermal stem cell stimulation aids (StemCell Technologies, Vancouver, Canada), including medium (MesenCult MSC basal medium) and antibiotics (Jin and Schuchman, 2003). The cells were cultured for 3 weeks and plastic-adherent cell populations (BM-MSCs) were used for the next experiment.

Example  3. Process of Transplanting Stem Cells into Mouse Brain

First the mice were anesthetized and fixed in stereotaxic, and 3 μl of stem cells obtained in Example 1 were aspirated with a Hamilton syringe.

Glass capillary (World Precision Instrument, Inc.) was cut out with diamond knives to the size that could be securely fixed to the end of the Hamilton syringe outlet.

The cut glass capillary was inserted into the Hamilton syringe outlet, and the hardener (Hardener) and the adhesive (Resin; Araldite Rapid) were mixed for 45 seconds, and then fixed on the joint within 4 minutes.

After confirming that the stem cells come out through the glass capillary (glass capillary), after fixing the hamilton syringe in a stereotaxic device (stereotaxic), stem cells were transplanted into the disease model mouse brain. At this time, 0.5 × 10 ⁶ cells in 3 μl of the cell suspension were transplanted at a rate of 0.15 μl / min.

Example  4. Confirmation of transplant

After grafting DsRed protein-expressing stem cells into the mouse brain using a cell injection syringe, the cerebellum of the transplanted mice and the cerebellum of mice not transplanted with DsRed protein-expressing stem cells were compared and observed by fluorescence microscopy. As shown in Figure 2, using the cell injection syringe of the present invention was able to observe the red fluorescence in the brain of the mouse transplanted with DsRed protein-expressing stem cells (Fig. 2A), thereby making the stem cells transplanted effectively I could confirm the loss.

The present invention relates to a cell injection syringe that minimizes damage to animal tissues when injecting cells in vivo and prevents cells injected into the holes due to the injection needles from flowing out again. By providing a method for transplantation into cells, organs and organs of an experimental animal, it is possible to provide useful information for developing a therapeutic method using cells.

Claims (6)

A syringe comprising a cylinder, a piston and an outlet, wherein the outlet of the syringe is connected by a capillary. The syringe for cell injection according to claim 1, wherein the capillary is made of glass. The syringe for cell injection according to claim 1, wherein the discharge port and the capillary are fixedly connected with an adhesive and / or a curing agent. The syringe for cell injection according to claim 1, comprising cells in the cylinder of the syringe. The syringe for cell injection according to claim 4, wherein the cells are stem cells. The cell injection syringe according to claim 4, wherein the content of cells contained in the cylinder of the syringe is in the range of 1 to 5 µl.

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