KR101333789B1 - Separating vials and their uses - Google Patents

Abstract

The present invention relates to a vial for centrifugation comprising a body (110), an exposure hole (112) formed as an air passage, and a lower packing (113). The vial for centrifugation according to the present invention flows air into the vial to form an air layer between a PRP layer and a red blood cell layer in the low part so that the PRP layer can be conveniently separated and re-mixing or reflux phenomenon of each demarcatirn can be prevented. The above characteristic is a unique technology compare to traditional PRP separation kits. In addition, by using the vial for centrifugation, minimum amount of demarcatirn such as RPR inside blood can be conveniently separated without accurate and skillful technology.

Description

Separating Vials and Their Uses

The present invention relates to a centrifugal vessel and its use.

The blood is used to comprehensively check the health state of the human body, and recently, the plasma, platelets, and blood cells are separated from the blood, and platelets and plasma, except for blood cells, are plated with blood plasma, which is diluted with platelets. Platelet Rich Plasma is used for a wide range of therapeutic purposes. For example, it is used in a wide range of therapeutic fields, such as wrinkles, pigment diseases, scars and other skin diseases such as dark circles, stretch marks, and sagging skin, cosmetic surgery, foot ulcers, and pain treatments, and researches in the field of use are continuing.

As such, blood has been widely used to separate blood from each component for various medical purposes. Among them, a blood centrifugation process using a specific gravity difference of blood components and a process using a specific composition are used. It is widely used. The centrifugation process is a process of performing delamination using the specific gravity difference of each component constituting the blood at a speed of about 3,000 rpm. When centrifuging the blood, the heaviest blood cells form a lower layer and leukocytes from above And platelet layers and plasma or serum layers thereon.

Conventionally, the separation of plasma, platelets, and blood cells from blood is performed by first collecting blood from a human body using a syringe, and then transferring the blood to at least the first and second centrifugation kits to carry out the centrifugation process using a centrifuge. Obtained by separation. In more detail, the blood sample collected from the human body is injected into a tube-type test tube, and the test tube into which the blood is injected is centrifuged using a centrifuge. When the centrifugation is completed, the red blood cells descend to the lower part of the test tube and the plasma is placed above the red blood cells. Next, insert the needle of the syringe into the test tube to inhale and extract the plasma located above the red blood cells. Next, centrifugation of the extracted plasma once again collects PRP at the bottom of the test tube and collects the rest of the plasma including PPP (Platelet poor plasma) above the PRP. The syringe is then used to inhale the remainder of the plasma, including the PPP, located above the test tube, discharge it from the test tube, and extract the remaining PRP into the test tube with a new syringe.

However, since the method of separating blood components using centrifugation requires separation of blood cells as a syringe mainly on the naked eye of the operator, PRP components flow out from the separated blood cells or blood cells are included in the PRP, so the purity of plasma is increased. There was a problem with falling. Particularly, when the centrifugal syringe is pressurized from the top, or when the piston rod of the storage syringe is pulled downward to apply negative pressure to draw the blood pressure down from the blood cells, the negative pressure is applied to the separated state and then released. Density flow due to density difference occurs in the blood, and the separated blood components are mixed again, and residues of blood cells may remain on the wall surface of the syringe cylinder from which blood cells are extracted, making it difficult to separate pure PPP or PRP. In this problem, the prior art additionally performs a cumbersome process of increasing the purity by removing the blood cells by separating the PRP and PPP separated.

In addition, the existing centrifuge has a problem that it is difficult to control the amount of blood cells or plasma to be extracted when extracting the blood cells to the outside by pressing the piston rod of the syringe. Therefore, it is difficult to reliably separate only the platelet region, that is, the PRP portion, which occupies a relatively small amount in the whole blood, and there is a problem in that it depends on the skill of the operator. In particular, when pushing up the separated blood layer by applying pressure from the bottom of the centrifuge container, a backflow phenomenon occurs due to the pressurization, and the separated blood components are mixed again. In this case, it is inconvenient to use a separate pressurization mechanism to minimize the backflow phenomenon.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors earnestly tried to develop a PRP separation kit that can easily separate PRP and minimize the loss of PRP. As a result, a centrifugal separation vessel was developed to prevent the backflow phenomenon that occurred when the PRP fraction was separated in the conventional PRP separation kit. The centrifugal container of the present invention has an exposed hole through which air can enter and exit, and a separate syringe is coupled to the upper part of the centrifugal container to generate pressure in an upward direction to allow air to flow into the exposed hole. The present inventors have completed the present invention by confirming that an air layer is generated between the PRP layer and the lower red blood cell layer by the inflow of air, so that the PRP layer can be separated more conveniently, and remixing or reflowing of each fraction can be prevented. It became.

It is therefore an object of the present invention to provide a centrifugal vessel.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the invention, the invention provides a centrifugal vessel comprising:

(A) the main body 110 is formed with a distribution pipe 111 which is accessible to the sample on the upper portion, the separation means for suction separation of the fraction of the sample is coupled to the upper portion of the main body 110;

(b) an exposure hole 112 formed in the upper portion of the main body 110 and formed to allow air access, wherein the air is sucked through the exposure hole opened when the fraction is inhaled by the separating means, thereby forming an air layer between the fractions of the sample; Is formed; And

(c) a lower packing (113) formed in the main body (110) and retractable within the main body.

The present inventors earnestly tried to develop a PRP separation kit that can easily separate PRP and minimize the loss of PRP. As a result, a centrifugal separation vessel was developed to prevent the backflow phenomenon that occurred when the PRP fraction was separated in the conventional PRP separation kit. The centrifugal container of the present invention has an exposed hole through which air can enter and exit, and a separate syringe is coupled to the upper part of the centrifugal container to generate pressure in an upward direction to allow air to flow into the exposed hole. The present inventors confirmed that an air layer is generated between the PRP layer and the lower red blood cell layer due to the inflow of air, so that the PRP layer can be separated more conveniently, and remixing or reflowing of each fraction can be prevented.

The present inventors pay attention to the fact that the conventional PRP separation method has a backflow phenomenon when collecting a buffy coat by inserting a needle at the top of the separation kit, and the PRP layer is remixed with another fraction layer, thereby preventing such a backflow phenomenon. A centrifugal vessel was developed. In case of separating PRP by using a conventional centrifugal container, red blood cells and the like are easily mixed in the process of inhaling PRP with a separate separator (for example, a syringe), and therefore, high concentration and skill are required for PRP separation. . Therefore, a lot of experience is required to increase the precision of the control when separating the PRP, the present inventors by forming an air layer between the PRP layer and the other fractionation layer centrifugal separation can conveniently separate only the PRP layer without requiring the above-described technical precision I came up with the courage.

The centrifugal vessel of the present invention can be applied to all samples that can be separated or concentrated by the density gradient centrifugation method, for example, biological samples, such as blood, plasma, cell culture fluid, etc., but is not limited thereto.

According to a preferred embodiment of the present invention, the sample which can be separated or concentrated by the centrifugation vessel is blood, and the fraction is PRP (platelet rich plasma). As used herein, the term “PRP (Platelet rich Plasma)” is used to centrifuge blood samples to remove red blood cells and to separate platelet and growth factor-rich buffy coat layers. It is a high concentration of platelets and growth factors in a single blood.

The centrifugal container 100 of the present invention may be manufactured in any shape known in the art, for example, a cylindrical shape, a cylindrical shape that becomes narrower in an upward direction, and a width in an upward direction. This narrower and can be manufactured in a cylindrical shape with a bottleneck on the top. According to a preferred embodiment of the present invention, the main body 110 has a shape in which the upper portion of the main body becomes narrower in an upward direction.

The main body 110 of the centrifugal container 100 of the present invention is formed with a distribution pipe 111 through which a sample can enter and exit, and an upper part of the main body 110 capable of separating and separating fractions of the sample by suction. Means can be combined. In addition, the exposure hole 112 is formed in the upper portion of the main body 110 and is formed to enable the air inlet, the air is sucked through the exposed exposure hole when the fraction is inhaled by the separation means is the air layer between the fraction of the sample Can be formed. The lower packing 113 is formed inside the main body 110 and is mounted to be able to move back and forth within the main body. The lower packing 113 may be formed of an elastic material and is preferably formed of a material including rubber.

According to a preferred embodiment of the present invention, the exposure hole 112 may be equipped with a filter 150 or a plug 150 that can be opened and closed to prevent contamination of the sample. The exposed hole 112 may have a hole or protruding shape. When the exposed hole 112 has a protruding shape, the exposed hole 112 may have an inner or outer protruding shape. In addition, when the exposed hole is a protruding shape, in order to minimize contamination of the sample it can be produced in a funnel shape that narrows the protruding portion. One or more exposed holes 112 may be formed in the centrifugal container of the present invention, preferably two or less, and even more preferably one.

According to a preferred embodiment of the present invention, the separating means coupled to the upper portion of the main body 110 is any means capable of inhaling and separating the fraction by combining to the upper portion of the main body 110, such as a syringe, pipette or dropper Although possible, it is most preferable to use a syringe when considering economics and efficiency. It is very convenient to use a syringe, and it will be possible to separate the fraction formed on the top of the body with precision. In this case it would be desirable to separate the fractions using a sterile syringe (syringe) to prevent contamination.

According to a preferred embodiment of the present invention, the centrifugal container 100 of the present invention may further include a connector 140 connecting the centrifugal container 100 and the separating means. As shown in FIG. 1, the lower centrifugal container 100 may be connected to a syringe, which is a separating means, through the connector 140. In the case of multiple fractionation layers, one or more syringes may be sequentially separated after connecting one or more syringes by connecting connectors.

When using the centrifugal container 100 of the present invention, a small amount of components contained in the sample can be separated out. That is, after centrifugation, the lower packing is pushed up so that the layer to be separated is positioned at the upper portion 111 of the main body, and the boundary of the fractionation layer is positioned near the exposure hole 112. This process may be possible by combining the syringe with the upper part of the body and then generating pressure in the upward direction to suction off the unnecessary fraction layer on the upper part, and also by applying physical pressure to the outside of the lower packing. Subsequently, after the new syringe is coupled to the upper portion of the main body, the exposure hole 112 is opened to allow air to flow into the main body 100. When the pressurization of the upward direction is again generated using the syringe, an air layer is formed between the fractionation layer to be separated and the unnecessary fraction layer below the air by flowing into the exposure hole 112. After the air layer is formed, the fractionation layer to be separated can be easily separated by a syringe.

The features and advantages of the present invention are summarized as follows:

(a) The present invention provides a centrifugal container including a main body 110, an exposure hole 112 and a lower packing 113 formed to allow air in and out.

(b) In the centrifugal separation vessel of the present invention, by introducing air into the vessel, an air layer is generated between the PRP layer and the lower red blood cell layer, so that the PRP layer can be more conveniently separated, and remixing or reflowing of each fraction is prevented. There is an advantage in that it corresponds to unique technical features compared to conventional PRP separation kits.

(c) In addition, when the centrifugal vessel of the present invention is used, very small fractions such as PRP in the blood can be conveniently separated without sophisticated and skilled techniques.

1 shows a perspective view of a centrifugal container, a separating means (syringe) and a connector according to a preferred embodiment of the present invention.
2 shows a cross-sectional view of a centrifugal vessel according to a preferred embodiment of the present invention.
Figure 3 schematically shows a method for separating fractionated blood according to a preferred embodiment of the present invention. (Iii) The left figure shows the state before centrifugation (A: blood). (Ii) The figure shows the state immediately after centrifugation (B: plasma, C: PRP, D: blood cell). (Iii) On the right side, the upper packing is pushed up by using the syringe coupled to the connector 140, and the lower packing is pushed up, and after the B part is separated, the upper part is again pressed to separate the C part. It shows after. It can be seen that an air layer is formed between the B and C layers.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Example 1: Preparation of Centrifuge Vessel

The main body 110 has a substantially cylindrical shape, and the main body 110 has a shape in which the upper portion of the main body becomes narrower in an upward direction. The upper portion of the main body 110 is formed with a distribution pipe 111 that can be sampled in and out, the upper portion of the main body 110 may be coupled to the separation means for suction separation of the fraction of the sample.

An exposed hole 112 is formed in the upper portion of the main body 110 to allow air to enter and out. When the fraction is inhaled by the separating means, the air is sucked through the exposed hole 112 to open the air layer between the fractions of the sample. This could be formed. The exposure hole 112 is equipped with a filter for preventing contamination of the sample, and the stopper 150 is mounted to enable the opening and closing of the exposure hole. The exposed hole was manufactured in the shape of an external protrusion so that the stopper can be coupled (see FIG. 1).

On the other hand, the inside of the main body 110 is equipped with a retractable lower packing 113, so that the sample can be pushed up (up) direction.

Since the main body 110 of the centrifuge container 100 of the present invention becomes narrower as the upper portion thereof goes upward, the volume of the upper and lower sections between the upper and lower portions of the main body 110 is smaller than that of the other portions of the red blood cells and the plasma. As the upper and lower thicknesses of the layers forming the boundary become larger, the boundary becomes more clearly visible.

Example 2 Separation of PRP Using Centrifuge Vessel

The anticoagulant was aspirated with a syringe before blood was drawn with the syringe. An appropriate amount of anticoagulant is about 10% of the amount of blood to be collected. Blood was collected from humans with a syringe inhaled with anticoagulant. Blood was injected into the body 110 through the distribution pipe 111 of the upper body. Next, the centrifuge was used to centrifuge the vessel of the present invention for about 5 minutes at a speed of 3000 rpm. At this time, the red blood cells are lowered to the lower part of the main body 110, and the plasma is located above the red blood cells. In the centrifuged state, a buffy coat layer excluding red blood cells and plasma is positioned at a central portion of the main body 110 due to individual differences such as age and gender of blood collection targets. The buffy coat layer refers to a pale yellow thin white blood cell layer formed on the red blood cell layer when the blood is centrifuged. When the blood is centrifuged, the red blood cell layer is located at the bottom, and the buffy coat layer is placed on the buffy coat layer, and the buffy coat layer is platelet rich plasma layer (PRP, platelet rich plasma) and platelet deficient plasma layer (PPP, platelet). poor plasma).

The PRP layer is very small and requires great care to separate it. The present inventors centrifuged using the centrifugal container 100 of the present invention (FIG. 3, middle), the connector 140 and the syringe (syringe) mounted on the top of the centrifugal container 100 layer B Was separated and removed. In this case, when pressurization occurs upward by the syringe, the lower packing 113 mounted inside the main body moves upward, and the B layer flows into the syringe. The syringe was pulled upward to remove the B layer, so that the C layer including the PRP layer was positioned in the space of the upper portion 114 of the main body, and the boundary portions of the C and D layers were close to the exposed holes. The upper 114 space of the body is narrower and smaller in volume than the lower space of the body, so that a small amount of PRP layer is easily seen.

Thereafter, the syringe coupled to the connector 140 was replaced with a new syringe, and the exposure hole 112 was opened to allow air to flow into the main body 100. The exposure hole 112 is equipped with a filter to prevent contamination. Subsequently, pressurization was again generated using a syringe, and air was introduced into the exposure hole 112 to form an air layer between the C and D layers. After the air layer was formed, the cap was closed again, and only the C layer (PRP) was separated and collected using a syringe.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Centrifuge Vessel: 100
Body: 110
Distribution line: (111)
Exposed Balls: (112)
Bottom Packing: (113)
Upper part inside the main unit: (114)
Extruded Holes: (115)
Syringe: 120, 130
Connector: 140
Plug: (150)
Syringe-type Centrifuge Containers: 160

Claims (9)

Centrifugal vessel 100 comprising:
(A) the main body 110 is formed with a distribution pipe 111 which is accessible to the sample on the upper portion, the separation means for suction separation of the fraction of the sample is coupled to the upper portion of the main body 110;
(b) an exposure hole 112 formed in the upper portion of the main body 110 and formed to allow air access, wherein the air is sucked through the exposure hole opened when the fraction is inhaled by the separating means, thereby forming an air layer between the fractions of the sample; Is formed; And
(c) a lower packing (113) formed in the main body (110) and retractable within the main body.
The centrifugal container according to claim 1, wherein the main body (110) has a shape in which the upper portion of the main body becomes narrower in an upward direction.
The centrifugal container according to claim 1, wherein said separating means is a syringe.
The centrifugal container according to claim 1, wherein the exposure hole (112) is equipped with a filter for preventing contamination of the sample.
The centrifugal container according to claim 1, wherein the exposure hole (112) is equipped with an opening and closing stopper (150).
The centrifugal container according to claim 1, wherein the shape of the exposed hole (112) is a hole, a protruding or an outer protruding shape.
The centrifugal container according to claim 1, wherein the centrifugal container (100) is in the shape of a syringe.
The centrifugal container according to claim 1, wherein the centrifugal container (100) further comprises a connector (140) connecting the centrifugal container (100) and the separating means.
The centrifugal container according to claim 1, wherein the sample is blood and the fraction is platelet rich plasma (PRP).

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