KR20150008548A - Extract kit of buffy coat - Google Patents

Abstract

The present invention relates to a buffy coat extracting kit used in extracting a buffy coat after extracting blood for an autohemotherapy skin reproduction procedure and a cell reproduction procedure. The buffy coat extracting kit comprises: a plasma unit (40) made of a transparent resin, serving as a place where a plasma layer is generated when blood is centrifuged, and formed in a cylindrical shape having a large diameter; a buffy coat unit (70) having a diameter gradually reduced towards the bottom of the plasma unit (40) and formed in a tube shape having a small diameter and enabling a buffy coat layer to be formed therein; an erythrocyte unit (80) having a vertical or horizontal support rib (60) formed in the outer surface of the plasma unit (40) or buffy coat unit (70) to have the same maximum diameter as the erythrocyte unit (80) or extend to the inner side surface of a centrifuge bucket, connected with the bottom of the buffy coat unit (70), and having an erythrocyte layer formed therein as a part which is directly inserted in the centrifuge bucket; an elastic bottom packing (90) formed at the bottom of the erythrocyte unit (80) and sealing the bottom of the kit body (10); a bottom cover (100) having an open center; and a top cover (20) coupled with the top end of the plasma unit (40) of the kit body (10) to prevent the leakage of blood, assembled with a top packing (30), and having an open center so that blood can be fed and a buffy coat can be extracted through a syringe.

Description

Extract kit of buffy coat

The present invention relates to a medical device, and more particularly, to a buffy coat extraction kit used for extracting a buffy coat by collecting blood for autologous blood skin regeneration and cell regeneration.

Separation of biological materials is the most fundamental work for cell culture and DNA identification and amplification as well as cell material analysis in all biological genetics and medicine. Currently, the world is competing in the field of regenerative medicine using stem cells in medicine. To obtain such stem cells, there are embryonic stem cells obtained from Blstocyst at the early stage of development and adult stem cells obtained from the adult or placenta after development.

Among these, there are three main ways of obtaining adult stem cells: first, through bone marrow; second, through umbilical cord blood; It can be easily obtained through blood (mesenchymal blood). All of these methods are first centrifuged to separate the components of hematopoietic platelet plasma into blood, and then isolate mononuclear cells from the stem cells.

In this process, we consider the conventional centrifugation methods as follows. In a solution containing various substances, especially when blood of human or animal is collected and centrifuged, the erythrocyte white blood cells platelets and other plasma components in the blood are separated from the bottom of the centrifuge container And the heavy one.

In the process of transferring the peripheral blood collected through the injection needle to the centrifuge vessel, the cap of the centrifuge vessel is opened and the needle of the syringe is placed inside the vessel for centrifuge. Then, the peripheral blood in the syringe is transferred to the centrifuge vessel After moving the needle, isolate the needle from the container for the centrifuge, close the open cap, insert the container for the centrifuge into the bucket of the centrifuge, set the desired time and rotational speed (rpm) or gravitational acceleration (g) .

Normally, red blood cells occupy the heaviest part of the blood and form in the lower part. In males, 47% of the total blood and 42% of the women. Above it, leukocytes and platelets account for about 1% of total blood, including monocytes, peripheral blood stem cells.

The remainder is the serum and fibrinogen called plasma on the upper layer. After centrifuging the centrifuge, take out the container and open the cap again to extract the desired substance out of the separated materials by the specific gravity difference of each constituent material through the pipette, capillary or needle, and then the remaining components are stored separately or discarded or recycled .

Recently, the autologous blood cell regeneration procedure (also referred to as "PRP (Platelet Rich Plasma) procedure" hereinafter referred to as "PRP procedure") has been spotlighted. Which is used to treat wound healing by regenerating cellular tissue by injecting the damaged tissue of the damaged tissue and extracting the damaged tissue. And so on.

It is known that platelets of blood are rich in growth factors. These platelets are known to multiply cells in the human body, produce collagen, regenerate new blood vessels and heal wounds.

Human blood contains about 100,000 to 200,000 platelets per microliter. To be effective, PRP should concentrate platelets to more than 1 million platelets per microliter. As a method of concentrating the platelets, a method of separating the collected blood into layers using a centrifuge is used. When the blood is separated into layers by a centrifugal separator, as shown in FIG. 1, A red blood cell layer is formed, and a yellow buffy coat layer consisting of white blood cells and platelets is formed on the red blood cell layer. The uppermost layer above the buffy coat layer is divided into a light yellow plasma layer composed of plasma.

The volume ratios of the separated blood layers are, on average, about 45% for the red blood cell layer, 1% for the buffy coat layer, and 54% for the plasma layer. In the PRP procedure, the buffer layer, which is rich in platelets, is extracted from the separated blood, and the proportion thereof is only 1%. Thus, only a BUFFY COAT can be obtained with a normal blood collection kit It is difficult to extract.

A centrifuge test tube for extracting whole blood from a patient and concentrating and fractionating platelets is disclosed in an assembly, a kit, and a method (applicant: ESTA TECHNOLOGIES LIMITED) for platelet-rich plasma formation This test tube for centrifugal separation has a cylindrical shape with a uniform inner diameter, which is difficult to accurately extract only the buffy coat as described above.

In the centrifugal separation kit and the centrifugal separation method using the same (Applicant: Glottec), whole blood or body fluids are sampled, and a syringe is installed in the centrifugal tube, A centrifugation kit is disclosed in which a substance (platelet-rich plasma or stem cells) is collected directly into a syringe to easily extract a target substance. The centrifugation kit also includes a centrifuge tube, which has a cylindrical inner diameter uniform, (BUFFY COAT), there is a disadvantage in that it is possible to collect separation gel or red blood cells in addition to the buffy coat. Thus, as shown in the applicant's patent publication 10-2012-0089723, the diameter of the centrifuge container A chamber is formed and a BUFFY COAT is formed in the chamber to separate the chamber from the upper chamber and the intermediate chamber, There has been proposed a buffy coat extraction kit which forms at least one rib on the side surface to prevent the phenomenon of flowing right and left during centrifugal separation in a centrifugal separator of an angle type in particular and precisely separates the substance contained in the container for the centrifugal separator, When the BUFFY COAT layer is accommodated in the lower chamber, there is a lack of means for controlling it.

In addition, the conventional kit forms a stopper and is directly opened by a human hand, injecting blood from a syringe and then closing it with a human hand, which is very unsanitary, and blood can be exposed to external air and infected There was concern.

Korea Patent No. 10-2010-0116106 (ESTA TECHNOLOGIES LIMITED) 2010.10.29 Korea Patent Publication No. 10-2011-0045980 (Glotech Co., Ltd.) 2011. 5. 4 Korea Patent Publication No. 10-2012-0089723 (Allen) June 8, 2012

In order to separate the biological cell material and especially the recent PRP treatment as described above, there is a need for a buffy coat extraction kit capable of easily extracting a BUFFY COAT in which blood platelets are concentrated in a centrifugal separator.

The buffy coat extraction kit for such a purpose should have a structure in which a BUFFY COAT layer can be easily identified in centrifuged blood and can be accurately extracted by a syringe.

Depending on the person and the same person in the blood, the composition ratio of red blood cells, buffy coat, and plasma is not uniform depending on the health state at the time of blood collection. In this way, the position of the buffy coat is always adjusted And extract it accurately.

In the process of extracting, it is a structure which can prevent unhygienic work which is exposed to outside air by being opened and closed by a person's hand, and is contaminated by being in contact with a human hand, but also can be easily handled with alcohol or the like The site to be inserted must be disinfected.

It is necessary to suppress the lateral flow of the kit as much as possible during centrifugal rotation so as to prevent disturbance of the buffy coat layer thereby to enable precise centrifugal separation.

The buffy coat extraction kit of the present invention is for accurately extracting the buffy coat of the buffy coat layer by injecting blood and separating it into layers of a red blood cell layer, a buffy coat layer and a plasma layer by using a centrifuge, And at least one or more pairs of support ribs 60 are formed on the outer surface of a part of the kit body 10. The upper packing 20, the upper packing 30, the lower packing 90, And support protrusions 50 are formed.

The method of extracting the buffy coat in blood using the buffy coat extraction kit according to the present invention is as follows. First, the exposed portion of the upper packing 20 is disinfected with alcohol cotton or the like, and then the upper packing 20 After the blood is poured into the buffy coat extraction kit and the syringe is removed, the buffy coat extraction kit is attached to the centrifuge to separate the blood into layers, and a BUFFY COAT layer is applied to the buffy coat 60 When the erythrocyte segment 80 is formed on the erythrocyte segment 80, the lower packing 90 is pushed up by a separate pin or piston so that the BUFFY COAT layer comes to the buffy coat portion 70,

At this time, a separate plunger or piston passes through the open structure of the lower plug 100 to push up the lower portion of the lower packing 90.

The lower packing 90 has a concave inner structure so that the piston or the piston coupled thereto can be pushed up and further a groove of a straight line or a cross is formed at the lower end so as to be coupled with a corresponding portion of the plunger or piston, So that the lower packing 90 can be pushed up stably without slippage.

Then, a step of piercing the needle tip of the syringe so as to reach the BUFFY COAT layer and extracting a BUFFY COAT is sequentially performed.

When the blood is centrifugally rotated using a centrifugal separator using the buffy coat extraction kit of the present invention, although a small amount of the buffy coat or buffy coat layer of about 1% of the whole blood is formed at a considerable height in the buffy coat portion, It is possible to extract it accurately.

The buffy coat extraction kit of the present invention is a kit for extracting buffy coats according to the present invention in which the lower packings are pushed up to bring the buffy coat layer to the buffy coat part even if the composition ratio of the red blood cells, the buffy coat and the plasma is not uniform, It is possible to extract it correctly.

The buffy coat extraction kit of the present invention is a closed system that does not open and close a cap, and is easily disinfected with an alcohol cotton or the like and then injects blood or extracts a buffy coat. When the cap is opened or the cap is opened and closed, The contamination which may be generated from the direct contact of the blood with the germs and the like in the blood is basically blocked, thereby increasing the storage period of the buffy coat.

The buffy coat extraction kit according to the present invention can prevent the deformation of the kit and the scattering of the buffy coat layer due to the fine lateral movement and high gravity acceleration caused by the centrifugal rotation due to the support ribs and the support protrusions.

In addition, the buffy coat extraction kit of the present invention can easily obtain the buffy coat in the busy hospital business by forming the buffy coat on the buffy coat part by simply pushing the lower packing by one centrifugal separation, thereby shortening the time.

In addition, the buffy coat extractor of the present invention prevents leakage of blood during centrifugation due to a strong adhesion force of the lower packing caused by a load applied during centrifugal separation.

Figure 1 shows the layered constituents of blood separated by a centrifuge.
2 is a partial enlarged view of one embodiment of a buffy coat extraction kit of the present invention.
3 is a cross-sectional view of one embodiment of a buffy coat extraction kit of the present invention.
FIG. 4 shows a step of extracting a buffy coat from blood collected using a buffy coat extraction kit according to the present invention.
Description of reference numerals used in the main parts of the drawings
The present invention relates to a kit comprising a kit body and an upper packing in which the upper packing is provided with an upper stopper and a blood plasma unit.

Referring to the drawings, the following will be described. FIG. 1 shows a layered component of blood separated by a centrifugal separator, and FIG. 2 is a partially enlarged view of one embodiment of a buffy coat extraction kit of the present invention. FIG. 3 is a cross-sectional view of an embodiment of a buffy coat extraction kit according to the present invention, and FIG. 4 shows a step of extracting a buffy coat from blood collected using a buffy coat extraction kit according to the present invention.

The buffy coat extraction kit of the present invention is used for accurately extracting the buffy coat of the buffy coat layer by injecting blood and separating it into layers of a red blood cell layer, a buffy coat layer and a plasma layer by using a centrifugal separator. 3, a kit body 10, an upper stopper 30, an upper packing 30, a lower packing 90, and a lower stopper 100 are formed.

The kit body 10 is made of a transparent resin as a cylinder for storing blood, and has a cylindrical plasma section 40 having a large inner diameter, and an inner diameter for forming a buffy coat layer formed continuously in the lower part of the plasma section 40 And a red blood cell layer formed on the lower part of the buffy coat part 70. The erythrocyte part 80 is a part directly inserted into the bucket of the centrifugal separator.

The lower packing 90 is coupled to the inside of the lower end of the erythrocyte section 80 and the lower stopper 100 is fastened to the lower end of the erythrocyte section 80 and the lower end of the plasma section 40 or the buffy coat section 70 And a support protrusion 50 formed on the outer side of the vertical or horizontal support rib 60 and the support rib 60 that is the same as the maximum outer diameter of the erythrocyte section 80 or on the inner side of the centrifuge bucket .

Normally, when a large amount of blood is injected, a large amount of buffy coat can be obtained. The buffy coat extraction kit is inserted and rotated in a bucket of a centrifuge. Since the inner volume of the bucket is limited but the outside is not limited, (40) can be made larger. At this time, since the buffy coat extraction kit is formed in a fragile shape, when the centrifuge is rotated, the lateral flow phenomenon becomes more severe, and a minute amount of the buffy coat layer is dispersed while being shaken, making it difficult to accurately collect and extract the buffy coat layer.

Therefore, the support ribs 60 are provided to prevent damage and deformation due to high gravity acceleration applied to the kit body 10 during centrifugation, and to minimize the lateral flow of the buffy coat extraction kit when the centrifuge is rotated. At least one horizontal support rib 60 or a pair of vertical support ribs 60 as shown in the figure are formed to surround the outer surface of the kit body 10 horizontally, Supporting protrusions 50 are formed on the outside of the ribs 60.

These centrifugal separators have different inner diameters for each centrifugal separator. When inserted into a bucket with a small diameter, they can be easily pushed while sliding while applying a slight force. When the diameter is slightly larger In the case of a bucket, it is inserted closer to the inner surface of the centrifuge bucket, thereby preventing the lateral flow of the buffy coat extraction kit caused by rotating the centrifuge, thereby preventing the buffy coat layer from being disturbed.

A protrusion is formed on the outer surface of the upper part of the plasma part 40 and a soft upper packing 20 such as silicone is formed in the upper part of the plasma part 40 so that the middle part is convex, And the upper stopper 30 forms a groove on the inner surface thereof to engage with the projection of the plasma section 40 to maintain the air tightness of the kit body 10. [

When the blood is centrifuged, the buffy coat layer is formed on the middle part of the buffy coat part 70 or at least the red blood cell part 80 when the blood body is centrifuged considering the composition ratio of erythrocytes, buffy coat, The diameter and length of the blood plasma section 40, the buffy coat section 70, and the erythrocyte section 80 are determined.

The connecting portion between the blood plasma section 40 and the buffy coat section 70 of the kit body 10 and between the buffy coat section 70 and the erythrocyte section 80 gradually changes in diameter, It is good.

The upper stopper 20 is formed with a middle open structure in which the convex portions of the upper packing 20 are coupled so that the exposed portion of the upper packing 20 is easily sterilized with alcohol or betadine before use Then it will be available.

The lower packing 90 is provided in the erythrocyte portion 70 and advances along the axial direction of the kit body 10, and may be formed as a plurality of layers. And is prevented from escaping to the outside by the lower stopper 100. The lower stopper 100 is formed to be fitted to the inner surface of the lower end of the erythrocyte section 80, and the diameter of the lower end of the lower stopper 100 is not larger than the outer diameter of the erythrocyte section 80. In this way, the outer diameter of the erythrocyte section 80 can be formed as close as possible to the inner diameter of the bucket, thereby ensuring a space for storing the maximum amount of blood in the erythrocyte section 80, The maximum amount of buffy coat can be obtained.

The lower stopper 100 can also be fused by ultrasonic welding or the like.

Although not shown, a screw thread is formed on the inner surface of the lower stopper 100, and a corresponding screw is formed on the outer circumferential surface of the piston or piston so as to be pushed up.

The above-described buffy coat extraction kit has a structure in which it is separated from the outside air. In the process of extracting the buffy coat by injecting blood from the outside, it prevents the exposure to the outside air to prevent the deterioration of the blood, can do.

Further, the lower stopper 100 is formed in a vertically-opened structure so that the lower packing 90 can be directly pushed up from the outside by a pusher, a piston, or the like.

The lower packing 90 is an elastic part directly receiving gravitational acceleration of blood generated during rotation of the centrifugal separator 90. The lower packing 90, which is an elastic body, is pressed by a strong gravity load, The diameter thereof is increased so as to be closer to the outside, that is, toward the inner circumferential surface of the erythrocyte portion 80 of the kit body 10, and blood is prevented from leaking to the outside.

The lower packing 90 may be recessed inwardly of the lower packing 90, and may be threaded inside, and then combined with a corresponding separate piston to move forward and backward.

On the other hand, the lower packing 90 is further formed with a groove of a straight line or a cross at the lower end thereof to engage with a corresponding portion of the push rod or the piston formed corresponding thereto, so that the lower packing 90 can be stably pushed up without slippage.

The procedure for extracting the buffy coat in the blood using the buffy coat extraction kit of the present invention as described above is as shown in FIG. 4. The blood is injected into the buffy coat extraction kit using a syringe as shown in FIG. 4 , The syringe is removed, and the buffy coat extraction kit is attached to the bucket of the centrifuge to separate the blood into layers.

4 (c), the buffy coat layer is formed on the buffy coat portion 60 of the body 10 of the buffy coat extraction kit, as shown in FIG. 4 (c), when the blood is separated into layers by the centrifugal separator as described above. do. Then, as shown in FIG. 4 (d), the buffy coat is extracted by making the needle tip of the syringe reach the buffy coat layer.

On the other hand, when the volume of the erythrocyte section 80 is formed to be large in the volume configuration of the kit body 10, the buffy coat layer is formed on the erythrocyte section 80 In this case, if the lower packing 90 is simply pushed up by a rod or a piston so that the buffy coat layer comes to the narrow buffy coat portion 70 as shown in FIG. 4 (c) The buffy coat layer is visible. Then, as shown in Fig. 4 (d), the buffy coat may be extracted by moving the needle tip of the syringe to the buffy coat layer.

As described above, the buffy coat extracted with the buffy coat extraction kit can separate the buffy coat very accurately, and a PRP of excellent quality can be obtained.

Claims (7)

A plasma part 40 made of a transparent resin and having a large inner diameter as a place where a blood plasma layer is formed when blood is centrifuged, and a buffy coat layer formed continuously in the lower part of the plasma part 40 The side wall of the blood plasma section 40 or the buffy coat section 70 has the same diameter as the maximum diameter of the red blood cell section 80 or extends to the inner side surface of the centrifuge bucket, A red blood cell portion 80 having a large inner diameter formed in a lower portion of the buffy coat portion 70 to form a red blood cell layer and a lower packing portion 60 for sealing the lower portion of the red blood cell portion 80, (90); < / RTI >
And is connected to the upper end of the plasma part 40 of the kit body 10 to prevent blood from leaking. The center part of the kit body 10 is opened and connected to the upper packing 20. The blood is injected using a syringe and the buffy coat is extracted An upper stopper (30) for allowing the upper stopper
And a buffy coat extracting kit 2. The kit according to claim 1, wherein a lower stopper (100) having an open structure is fastened to the lower portion of the kit body. The buffy coat extracting kit according to any one of claims 1 to 2, characterized in that the outer diameter of the lower stopper (100) does not exceed the diameter of the erythrocyte part (80) The buffy coat extracting kit according to any one of claims 1 to 3, characterized in that a screw thread is formed on the inner surface of the lower stopper (100) The buffy coat extracting kit according to claim 1, characterized in that the supporting ribs (60) are formed horizontally or vertically on the plasma part (40) or the buffy coat part (70) 6. The buffy coat extracting kit according to any one of claims 1 to 5, characterized in that a support projection (50) is formed on the support rib (60) The buffy coat extracting kit according to claim 1, characterized in that the lower packing (90) is formed with a groove or a cross,

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