RU2698723C1 - Sterile system for blood separation and a method for producing thrombocyte enriched whole blood plasma - Google Patents

Abstract

FIELD: medicine; technological processes.

SUBSTANCE: group of inventions relates to the field of blood separation, in particular to the production of whole blood enriched thrombocyte plasma. Blood separation system comprises a flask adapted for installation in a centrifuge made of a transparent polymer and consisting of three interconnected parts – an upper cylindrical part with a cover, smoothly forming a hourglass shape passing through a narrow isthmus to a lower cylindrical part, wherein at the point of transition of the upper part of the bulb into the isthmus there are four stiffening ribs, and the surface of the bulb is equipped with information marks, differing by the fact that it is additionally equipped in the lower part with a rotating piston with a seal and a key-support connected to the piston grooves. At that, on the piston and on the inner surface of the base of the lower cylindrical part of the bulb having the appropriate bulge, there is a single-thread thread, four stiffening ribs are made elongated throughout the narrow neck is to the lower cylindrical part of the bulb, information marks are made in the form of five concentric thickenings on the bulb surface, note here that Label 1 is mark of piston initial position, Label 2, Label 3 and Label 4 are hematocrit markers. Label 4 is also a marker of leukotrombocyte layer (LTL) indicating level of technical availability beginning of leukotrombocyte cell suspension intake using any blood sampling syringe with needle, length of which is 40 mm, Label 5 (MAX mark) is made to indicate the maximum filling of the bulb with blood. Also disclosed is a method of producing thrombocyte enriched whole blood plasma when using the disclosed system.

EFFECT: group of inventions provides higher efficiency of producing thrombocyte enriched whole blood plasma with sufficient mechanical strength, ease of use and reliable functioning of the system for blood separation.

8 cl, 17 dwg

Description

The present invention relates to medical devices, is intended for the separation of blood and, in particular, to a method for producing platelet-rich plasma from whole blood using the proposed device.

A device and method for producing platelet-rich plasma is known, in which the process of its production is carried out using a test tube that provides an extension filter device equipped with a filter attached to the first end section and the tube. The filter of the device has an effective pore size that allows platelets to pass through and prevent the passage of large blood components (Patent KR 20100116106 (A), IPC A61M 1/36, A61M 37/00, 2010-10-29, Patent KR 20100041436 (A), IPC A61K 35/16, 2010-04-22).

Also known is a device and method for producing highly concentrated plasma from whole blood, consisting of: a syringe; tubes attached to the syringe to centrifuge the whole blood in the first syringe; and a membrane filter for removing water or a water-containing component. The membrane filter contains: a housing containing an inlet for connecting a syringe, a membrane, a second space formed in the housing into which plasma for filtration is introduced, a highly concentrated plasma containing components having a size larger than a given particle diameter and not passing through the membrane remains second space. (Patent RU 2578418 C2, IPC A61M 1/34, B01D 61/18, 02/03/2014).

Other devices and methods are also known for producing platelet-rich plasma with the release of the leukotrombocyte layer (LTS) of cells from whole blood. These products are in the form of an hourglass with a narrow neck, equipped with a cap and a piston in different versions (test tube "I-STEM PRP KIT":

https://www.youtube.com/watch?time_continue=12&v=2ypBh45-SV4, test tube "Ycellbio-kit": https://www.youtube.com/watch?v=y-mzQh0xRzk).

However, these devices do not provide the achieved platelet concentration in a fixed volume of platelet-rich plasma, do not have sufficient mechanical strength, reliability and ease of use in comparison with the proposed system for blood separation.

The technical goal of the proposed technical solution is: increasing the efficiency of the method of obtaining platelet-rich plasma from whole blood with sufficient mechanical strength, ease of use and the reliability of the proposed system for the separation of blood.

This goal is achieved by the fact that the proposed system for separation of blood contains a flask providing for the possibility of a threaded piston stroke, the piston itself, equipped with a sealant, a cap fixed on the flask and an attached key-stand. The lower part of the flask at its base has a thickening, while the inner surface of the wall of the flask in this section has a single start thread, which allows the threaded stroke of the piston. The middle part of the flask is a narrow isthmus, passing into the upper part of the flask. The transition point of the lower part of the flask to the isthmus, the isthmus itself and the place of its transition to the upper part of the flask is equipped with four stiffeners to increase mechanical strength. The surface of the system is additionally equipped with information marks, which are concentric thickenings on the surface of the flask, with Label 1 being the mark of the initial position of the piston, Label 2, Label 3 and Label 4 are the hematocrit labels, Label 4 is also the label of the platelet layer (LTS), indicating the level of the beginning of the technical feasibility of taking leukotrombocyte suspension of cells using any syringe with a needle, the length of which is 40 mm (they are not components of the system). Label 5 (Label MAX) is made to indicate the boundary of filling the system with blood.

When filling the system with blood to the boundary of Tag 5 (Tag MAX), its working volume is 15.0 ml. The flask of the system is made of a transparent polymer to enable visual control of filling the flask with blood before centrifugation, to assess the desired result after centrifugation, and the stages of collection of platelet-rich plasma. The volume of the lower part of the flask from Label 1 to the isthmus is 6.87 ml., The volume of the isthmus is 0.4 ml., The volume of the upper part of the flask from the isthmus to Label 5 (Mark MAX) is 7.73 ml. The outer surface of the upper edge of the flask is provided with a three-way thread for fixing the cap.

A method for separating blood in a sterile system using the above device includes the steps of:

- a needle with a length of 40 mm or more, mounted on the cone of the used syringe containing venous blood (not part of the system), is inserted all the way through the opening of the cap so that it is centered in the neck of the flask. Moreover, in the initial position, the upper edge of the piston seal is at the level of Mark 1, and the system itself can be installed on a key stand fixed in the grooves of the piston, or in the hands of the operator;

- from the used syringe, blood is injected into the system until reaching Mark 5 (MAX mark), which limits the initial working volume of the system to 15.0 ml .;

- the needle is removed from the system. The opening of the cover is closed with a plug, after which the key-stand is removed from the piston. The system is ready for centrifugation;

- the system is placed in a centrifuge;

- the centrifugation process is carried out at a rotation speed of 3000 to 4000 rpm. before separation of blood into fractions (erythrocyte mass, leukotrombocyte suspension of cells, blood plasma) and allocation of a leukotrombocyte layer (LTS) visible to the naked eye;

- the system is removed from the centrifuge when blood separation is achieved and visible LTS is obtained;

- the system is installed on a key-stand, thereby ensuring its stable position when resting on a smooth surface;

- depending on the level of location of the LTS, by rotating the key-stand in the counterclockwise or counterclockwise direction, we control the threaded stroke of the piston in order to level the level of the lower boundary of the LTS on Mark 4 (Label LTS);

- using the tab of the plug, the latter is removed from the opening of the lid;

- a needle, the length of which is 40 mm, put on the cone of the syringe (not part of the system), is inserted all the way through the opening of the cap so that the tip of the needle is at the level of Mark 4 (Label LTS) and is centered in the neck of the flask;

- the required volume of platelet-rich plasma is taken with a syringe.

The objectives described above, characteristic features and advantages of the present invention will become more clearly understood from the following detailed description, accompanied by the accompanying graphic materials on which are shown:

FIG. 1. Sterile system for blood separation (complete).

FIG. 2. The flask.

FIG. 3. Flask (general view).

FIG. 4. Piston (side view).

FIG. 5. Piston (bottom view).

FIG. 6. Piston (general view).

FIG. 7. Sealant.

FIG. 8. Sealant (general view).

FIG. 9. The cover.

FIG. 10. Cover (general view). A - plug (27) in a free state; B -

the hole (25) of the cover (5) is closed by a plug (27).

FIG. 11. Key stand.

FIG. 12. Key-stand (general view).

FIG. 13. General appearance of the system.

FIG. 14. Stage 1 of work with the system (1) (preparation).

FIG. 15. Stage 1 of work with the system (1) (the system is ready for centrifugation).

FIG. 16. Stage 2 of work with the system (1) (centrifugation).

FIG. 17. Stage 3 of work with the system (adjustment of the lower level

LTS borders).

The system (1) for separating blood contains a flask (2) made of a transparent polymer and consisting of three interconnected parts - the upper cylindrical part (11) with the lid (5), smoothly forming an hourglass shape passing through a narrow isthmus ( 10) into the lower cylindrical part (7), and at the place of transition of the upper part (11) of the flask (2) to the isthmus (10), four stiffeners (12) are made, and the surface of the flask is provided with information marks.

The system is additionally equipped at the bottom with a rotating piston (3) with a seal (4) and a key-stand (6) connected to the grooves (16 and 17) of the piston (3), while on the piston (3) and on the inner surface of the base the cylindrical part (7) of the flask (2) having a corresponding thickening (8), made a single thread (9). Four stiffeners (12) are made elongated along the entire length of the narrow isthmus (10) to the lower cylindrical part (7) of the bulb (2).

Information marks are made in the form of five concentric bulges on the surface of the flask (2), with Label 1 - marking the initial position of the piston, Label 2, Label 3 and Label 4 - hematocrit marks, and Label 4 is also a marking of the platelet layer (LTS), indicating the level of the beginning of the technical feasibility of taking leukotrombocyte suspension of cells using any syringe for blood sampling with a needle whose length is 40 mm, Mark 5 (Mark MAX) is made to indicate the boundary of the maximum filling of the flask (2) with blood.

The working volume of the flask (2) is 15.0 ml. when the system is filled with blood to the boundary of Label 5. The increased working volume of the flask (2) (outlined by labels), which allows to form a larger volume of the leukotrombocyte layer (LTS) of cells during centrifugation, is made with the possibility of increasing the final concentration of platelets in a fixed volume of the obtained platelet-rich plasma. The volume of the lower part (7) of the flask (2) from Label 1 to the isthmus (10) is 6.87 ml., The volume of the isthmus (10) is 0.4 ml., The volume of the upper part (11) of the flask from the isthmus (10) to Mark 5 is - 7.73 ml.

The outer surface of the upper edge of the flask (2) is provided with a three-way thread (13) for fixing the cap (5). A universal opening (25) with a plug (27) and a tongue (31) is made in the lid (5) for introducing blood before centrifugation and collection of plasma enriched with platelets after. The multi-function key-stand (6) is made with the possibility of reliable installation of the system on a smooth surface by fixing the key-stand (6) in the grooves (16, 17) of the piston (3).

The method of obtaining platelet-rich plasma from whole blood using the proposed system for the separation of blood is as follows:

- a needle (not shown), the length of which is 40 mm or more, put on the cone of the used syringe (are not parts of the system) containing venous blood, is inserted all the way through the universal hole (25) of the cap (5) so that it was centered in the neck (10) of the flask (2), while in the initial position the upper edge of the seal (4) of the piston (3) is at the level of Mark 1 of the flask (2), and the system (1) can be installed on the key stand (6) fixed in the grooves (16, 17) of the piston (3), or is in the hands of the operator;

- from the used syringe (not shown), blood is introduced into the system (1) until reaching Mark 5 (MAX mark), which limits the initial working volume of the system (1), equal to 15.0 ml .;

- the needle (not shown) is removed from the system (1), the hole (25) of the cover (5) is closed by a plug (27), after which the stand key (6) is removed from the piston (3), while the system (1) is ready for centrifugation;

- system (1) is placed in a centrifuge (not shown);

- the centrifugation process is carried out at a rotation speed of 3000 to 4000 rpm. before separation of blood into fractions (erythrocyte mass, leukotrombocyte suspension of cells, blood plasma) and allocation of a leukotrombocyte layer (LTS) visible to the naked eye;

- system (1) is removed from the centrifuge when blood separation is achieved and visible LTS is obtained;

- the system (1) is installed on a key-stand (6), thereby ensuring its stable position when resting on a smooth surface;

- depending on the location of the LTS, by rotating the key-stand (6) clockwise or counterclockwise, the threaded piston stroke (3) is controlled to align the level of the lower boundary of the LTS on Mark 4 (LTS Mark);

- using the tab (31) of the plug (27), the latter is removed from the hole (25) of the cover (5);

- a needle (not shown), the length of which is 40 mm, put on the cone of the syringe (are not parts of the system), is inserted all the way through the hole (25) of the cover (5) so that the tip of the needle is at the level of Mark 4 (Mark LTS) and was centered in the isthmus (10) of the flask (2);

- the required volume of platelet-rich plasma is taken with a syringe.

Thus, the working volume of system (1), equal to 15.0 ml, is within the boundaries outlined by Tag 1 and Tag 5.

The dimensions of system (1) are compatible with standard centrifuges used in clinical laboratory practice (example: Armed centrifuge: 80-2S).

System (1) provides for the possibility of introducing blood into it from the outside using any syringe with a volume of 20 ml with a needle, the length of which is 40 mm or more (they are not components of the system).

The flask (2) (Fig. 2, Fig. 3) of the system (1) is made of a transparent polymer, which makes it possible to visually control the filling of the flask (2) with blood before centrifugation, evaluate the desired result after centrifugation, and the stages of collection of platelet-rich plasma. The lower part (7) of the flask (2) at its base has a thickening (8), the inner surface of the wall of the flask (2) in this section has a single start thread (9), which allows the threaded stroke of the piston (3).

The middle part of the flask (2) is a narrow isthmus (10), passing into the upper part (11) of the flask (2). The transition point of the lower part (7) of the flask (2) to the isthmus (10), the isthmus (10) itself and the place of its transition to the upper part (11) of the flask (2) is equipped with four stiffeners (12) to increase mechanical strength. The volume of the lower part (7) of the flask (2) from Label 1 to the isthmus (10) is 6.87 ml. The volume of the isthmus (10) is 0.4 ml. The volume of the upper part (11) of the flask from the isthmus to Mark 5 (Mark MAX) is - 7.73 ml. The outer surface of the upper edge of the flask (2) is provided with a three-way thread (13) for fixing the cap (5).

The piston (3) (Fig. 4, Fig. 5, Fig. 6) of the system (1) is made of polymer. The outer surface of the lower part (14) of the piston (3) is provided with a single-thread (15), which enables the threaded stroke of the piston (3) along the thread (9) of the lower part (7) of the bulb (2). The working interval of the threaded stroke for the piston (3) is: forward stroke - 8 mm, which reduces the working volume of the system (1) by 1.8 ml; the back stroke is 2.7 mm, which allows to increase the working volume of the system (1) by 0.6 ml. The base of the lower part (14) of the piston (3) is a cross-shaped groove (16) (Fig. 5), continuing into a conical groove (17). Both grooves serve for connection and centering in the piston (3) of the key-stand (6), the rotation of which clockwise and counterclockwise drives the piston (3), thereby providing the ability to control the working volume of the system (1). The upper part (18) of the piston has a disk-shaped (19) and cross-shaped (20) protrusions for fixing on the piston (3) of the seal (4).

The seal (4) (Fig. 7, Fig. 8) of the piston (3) is made of polymer. The inner surface of the seal (4) has a concentric thickening looking inward - a shoulder (21), the inner diameter of which is smaller than the outer diameter of the disk-shaped (19) piston thickening, which ensures reliable fixation of the disk-shaped (19) piston thickening (3) in the groove (22). The outer surface of the seal has several concentric bulges (23), the outer diameter of which in a free state slightly exceeds the inner diameter of the housing of the lower part (7) of the bulb (2), which ensures a tight fit of the seal (4) and the piston seal (3) in the bulb body ( 2), causing the tightness of the connection.

The cover (5) (Fig. 9, Fig. 10) is made of polymer. The inner surface of the lid (5) is provided with a three-way thread (24), which secures the lid to the thread (13) on the upper part (11) of the flask (2). A hole (25) is provided in the center of the lid, the surface of which has a concentric groove-recess (26) to ensure reliable fixation of the plug (27), which in turn has a coaxial concentric thickening (28) on its outer surface. In addition, the outer surface of the plug (27) has an additional concentric thickening (29), the outer diameter of which in the free state slightly exceeds the diameter of the hole (25) of the cover (5), which ensures a tight fit of the plug (27) and its seal in the cover body ( 5), causing the tightness of the connection. The cap (27) is an integral part of the cover (5) connected to it by a leash (30). For ease of grip with your fingers, the plug (27) is equipped with a tongue (31).

The key stand (6) (Fig. 11, Fig. 12) is made of polymer. The base (32) of the key-stand (6) has the shape of a four-beam cross (33), the diameter of which exceeds the diameter of the upper part (11) of the bulb (2), which ensures the stability of the system (1) when resting on a smooth surface. The upper part (34) of the key-stand (6) is a cone-shaped protrusion-spike (35), bearing on its outer surface four coaxial protrusions (36), the dimensions and direction of which correspond to the grooves (16 and 17) of the piston (3), which determines the alignment of the key-stand (6) in the piston (3), as well as the ability to control its threaded stroke.

System (1) is supplied in an individual sterile package and is intended for single use.

The length of the system (1) (Fig. 1) with the closed cap (6) of the cover (5) is 118.47 mm. The initial working volume of the system (1) - the volume limited by Tag 1 and Tag 5 (Fig. 14) is 15.0 ml.

Stages of preparation for work (performed independently and independently of system (1), and the medical devices and excipients used in this process are not components of system (1)):

1. We make a fence of 1.5 ml. blood preservative in a syringe (not shown) with a volume of 20 ml;

2. Using the same syringe, we take the patient's venous blood to the level of 15 ml;

3. We mix the blood and the blood preservative making “swaying” movements with the used syringe (not shown).

Stages of working with the system (1):

Stage 1 (preparation):

Insert the needle, the length of which is 40 mm or more, put on the cone of the used syringe (not shown) with a volume of 20 ml (not part of the system) into the hole (25) of the cover (5) so that the needle tip is at the level Label 4 and was centered in the isthmus (10) of the flask (2) (Fig. 14).

In the initial position, the upper edge of the seal (4) of the piston (3) is at the level of Mark 1 of the bulb (2). In this case, the system (1) can be installed on the key stand (6), fixed in the grooves (16, 17) of the piston (3) (Fig. 14), or is in the hands of the operator.

We introduce blood from the used syringe into the system (1) until reaching Mark 5 (MAX mark), which limits the initial working volume of the system (1), equal to 15 ml.

We remove the needle from the system (1) and close the hole (25) of the cover (5) with a plug (27) (Fig. 10B), and then remove the stand key (6) from the piston (3). The system is ready for centrifugation (Fig. 15).

Stage 2 (centrifugation):

We place the system (1) in a centrifuge (Fig. 16). Note: the dimensions of system (1) are compatible with standard centrifuges used in clinical laboratory practice (example: Armed centrifuge: 80-2S).

Depending on the type of centrifuge, the centrifugation process is carried out at a speed of rotation from 3000 to 4000 rpm. before separation of blood into fractions (erythrocyte mass, leukotrombocyte suspension of cells, blood plasma) and allocation of a leukotrombocyte layer (LTS) visible to the naked eye. The average centrifugation time is 4 minutes, and in case of insufficient blood separation and the absence of visible LTS, additional time can be added, sufficient to obtain the desired result (the duration of centrifugation is determined by the achievement of the above result).

Stage 3 (collection of platelet-rich plasma):

We take out the system (1) from the centrifuge (it is not an integral part of the system). After making sure of the separation of blood and obtaining visible LTS, we install the system (1) on the key-stand (6), thereby ensuring its stable position when resting on a smooth surface. Depending on the location of the LTS, by rotating the key-stand (6) clockwise or counterclockwise (Fig. 17) we control the threaded piston stroke (3) in order to align the level of the lower boundary of the LTS on Mark 4 (Mark LTS). The working interval of the threaded piston stroke (3) is: forward stroke - 8 mm, which allows to reduce the working volume of the system (1) by 1.8 ml (Fig. 17 ΔB); the backward stroke is 2.7 mm, which allows to increase the working volume of the system (1) by 0.6 ml (Fig. 17 ΔВ).

Using the tongue (31), remove the plug (27) from the hole (25) of the cover (5). Insert the needle, the length of which is 40 mm, put on the cone of the syringe (not part of the system) into the hole (25) of the cover (5) so that the needle tip is at the level of Mark 4 and centered in the neck (10) flasks (2). We take the required volume of platelet-rich plasma with a syringe.

Thus, the proposed system for the separation of blood provides the following advantages when compared with known analogues:

1. A new mechanism for adjusting the working volume of the product has been created, which allows you to adjust the position of the lower boundary of the obtained LTS, which makes it convenient to collect platelet-rich plasma by using a syringe with a standard needle, the length of which is 40 mm or more;

2. The final platelet concentration in the fixed volume of the obtained platelet-rich plasma (1.5 - 2.0 ml) was increased due to the increased working volume of the product (outlined by tags) from 13.5 mm (in the case of the Ycellbio-kit tube) to 15.0 ml (in the case of the proposed system for blood separation) and the possibility of the initial formation of a larger in volume leukotrombocyte layer (LTS) cells during centrifugation;

3. Increased mechanical strength of the product due to the presence of four stiffening ribs throughout the narrow neck of the flask. In the case of the “Ycellbio-kit” test tube, stiffeners are provided only in its upper part;

4. Improved ergonomics and ease of use of the product, including due to the presence of a key-stand, as well as a universal hole in the cap for blood supply before centrifugation and collection of platelet-rich plasma after (in the case of the Ycellbio-kit tube, 2 holes are provided);

5. The issue of reliable installation of the product on a smooth surface has been resolved (in the case of the Ycellbio-kit tube, there is no stand).

Thus, the stated technical goal of the proposed technical solution is achieved: increasing the efficiency of the method of obtaining platelet-rich plasma from whole blood with sufficient mechanical strength, ease of use and the reliability of the proposed system for blood separation.

Claims (19)

1. System (1) for the separation of blood, containing a flask (2), adapted for placement in a centrifuge, made of a transparent polymer and consisting of three interconnected parts - the upper cylindrical part (11) with the lid (5), smoothly forming hourglass shape passing through a narrow isthmus (10) to the lower cylindrical part (7), and at the place of transition of the upper part (11) of the flask (2) to the isthmus (10) four stiffeners (12) are made, and the surface of the flask is equipped with information tags, characterized in that it is additionally provided and in the lower part a rotating piston (3) with a sealant (4) and a key-stand (6) connected to the grooves (16 and 17) of the piston (3), while on the piston (3) and on the inner surface of the base of the lower cylindrical part (7) a flask (2) having a corresponding thickening (8), a single start thread (9) was made, four stiffeners (12) of stiffness were made elongated along the entire length of the narrow neck (10) to the lower cylindrical part (7) of the flask (2), information marks are made in the form of five concentric thickenings on the surface of the flask (2), while Label 1 is the initial mark the position of the piston, Label 2, Label 3 and Label 4 are hematocrit labels, and Label 4 is also a label of the leukotrombocyte layer (LTS), indicating the level of the beginning of the technical feasibility of sampling leukotrombocyte suspension of cells when using any syringe for blood sampling with a needle whose length is 40 mm, Mark 5 (Mark MAX) is made to indicate the boundary of the maximum filling of the flask (2) with blood. 2. The system according to claim 1, characterized in that the working volume of the flask (2) is 15.0 ml when filling the system with blood to the border of Tag 5. 3. The system according to claim 1, characterized in that the working volume of the flask (2), outlined by labels, is selected with the possibility of forming a leukotrombocyte layer (LTS) of cells visible to the naked eye during centrifugation and obtaining an increased platelet concentration in a fixed volume of platelet-rich plasma. 4. The system according to claim 1, characterized in that the volume of the lower part (7) of the flask (2) from Label 1 to the isthmus (10) is 6.87 ml, the volume of the isthmus (10) is 0.4 ml, the volume of the upper part (11) the flask from the isthmus (10) to Label 5 is - 7.73 ml. 5. The system according to claim 1, characterized in that the outer surface of the upper edge of the bulb (2) is provided with a three-way thread (13) for fixing the cap (5). 6. The system according to claim 1, characterized in that a universal opening (25) is made in the lid (5) with a plug (27) and a tongue (31) for introducing blood before centrifugation and collection of plasma enriched with platelets after. 7. The system according to claim 1, characterized in that the multifunctional key-stand (6) is configured to reliably install the system on a smooth surface by fixing the key-stand (6) in the grooves (16, 17) of the piston (3). 8. The method of obtaining platelet-rich plasma from whole blood when using the proposed system for blood separation according to claim 1 includes the steps in which: - a needle, the length of which is 40 mm or more, put on the cone of the used syringe containing venous blood, is inserted all the way through the universal hole (25) of the cap (5) so that it is centered in the neck (10) of the flask (2) in this case, in the initial position, the upper edge of the seal (4) of the piston (3) is at the level of Tag 1 of the bulb (2), and the system (1) itself can be installed on the key stand (6), fixed in the grooves (16, 17 ) piston (3), or is in the hands of the operator; - from the used syringe, blood is introduced into the system (1) until reaching Tag 5 (Tag MAX), which limits the initial working volume of the system (1), equal to 15.0 ml; - the needle is removed from the system (1), the hole (25) of the cover (5) is closed by a plug (27), after which the stand key (6) is removed from the piston (3), while the system (1) is ready for centrifugation; - system (1) is placed in a centrifuge; - a centrifugation process is performed at a speed of rotation from 3000 to 4000 rpm until the blood is divided into fractions (erythrocyte mass, leukotrombocyte suspension of cells, blood plasma) and the leukotrombocyte layer (LTS) is visible to the naked eye; - system (1) is removed from the centrifuge when blood separation is achieved and visible LTS is obtained; - the system (1) is installed on a key-stand (6), thereby ensuring its stable position when resting on a smooth surface; - depending on the location of the LTS, by rotating the key-stand (6) clockwise or counterclockwise, the threaded piston stroke (3) is controlled to align the level of the lower boundary of the LTS on Mark 4 (LTS Mark); - using the tab (31) of the plug (27), the latter is removed from the hole (25) of the cover (5); - a needle, the length of which is 40 mm, put on the cone of the syringe, is inserted all the way through the hole (25) of the cap (5) so that the tip of the needle is at the level of Mark 4 (Label LTS) and is centered in the neck of the flask (10) (2); - the required volume of platelet-rich plasma is taken with a syringe.

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