Apparatus for separating and recovering blood components

Abstract

The invention relates to a device for separating and recovering blood components by means of two conical chambers which communicate via the base of the conical chambers, the first chamber having a conduit for supplying the fluid to be treated and means for recovering at least one component, the second chamber comprising a flexible membrane which is elastically deformable and a bag, the flexible membrane separating laterally the space open to the conduit for communication with the second chamber, and the volume of the bag varying according to the deformation of the membrane.

Description

Technical Field

[0001] This invention relates to the automated processing and separation of biological cells, such as those found in whole blood, and more specifically to a closed functional system capable of extracting cell populations, such as hematopoietic stem cell populations, for immediate use or for mixing with additive or storage solutions for subsequent separation and storage operations, and for use in processes that allow such extraction. The aim is particularly to automatically isolate the erythrocyte sedimentation rate (ESR) layer containing a large number of hematopoietic and mesenchymal stem cells.

[0002] More specifically, the present invention relates to the field of separating and recovering blood components, particularly lymphocytes and other white blood cells, by centrifugation.

[0003] Specifically, this separation from blood samples can be achieved by creating a density gradient above a fluid layer with a reference density in the centrifuge tube, such as a liquid polyfluorocarbon with a density of 1.077.

[0004] One application of these, particularly widespread in Asia, involves cosmetic surgery that uses an hourglass tube with two openings, subjecting it to centrifugation to prepare platelet-rich plasma (PRP). The erythrocyte sedimentation rate (ESR) layer is then collected in the neck region of the tube.

[0005] PRP is centrifuged to remove red blood cells and separate platelets and platelet-rich plasma (ERP) containing platelets and growth factors (erythrocyte sedimentation rate). In the surgical field, PRP has the function of reconstructing and repairing damaged parts of blood vessels. Growth factors that promote skin or tissue remodeling or regeneration (activating ligaments, tendons) can be concentrated for the treatment of arthritis, chronic back pain, pelvic pain, and shoulder and knee ligament injuries.

[0006] The consumable is in the form of a biconical tube, the two tubes being connected at their bases, and one of the tubes including a well-like portion on its outer surface, the well-like portion being configured to supply the collected blood. After centrifugation, the required amount of plasma for subsequently collecting growth factors present in the alpha particles of platelets, such as TGF-β (“transforming growth factor β”), VEGF (“vascular endothelial growth factor”), and PDGF (“platelet-derived growth factor”), is collected via the same opening. Background Technology

[0007] In the prior art, U.S. patent application US20160129438 is known, which describes a cup-shaped device for collecting biological fluids, which can be used with a centrifuge to separate the biological fluids into their constituent components. The cup-shaped device is hourglass-shaped, having an open upper portion and a lower portion, and a narrow central portion. A piston is slidably inserted into the lower portion, and a lateral orifice is provided for extracting liquid from the narrow portion.

[0008] Application TW201405127A is also known, which describes a structural component comprising multiple bodies. One of these bodies includes a first receiving compartment with a fixed volume and a second compartment with a variable volume, the two compartments being connected by a narrow portion that defines a communication path for establishing communication between the fixed-volume compartment and the variable-volume compartment.

[0009] Furthermore, patent application US3911918 is known, which describes a container made of a flexible plastic material for storing viable fluids such as blood. The container is formed as a single piece and has two storage compartments connected in fluid communication via the neck portion of a delivery conduit. One of the storage compartments has approximately 60% of its dimensions intended to hold a predetermined amount of blood, and the other storage compartment and the neck portion of the delivery conduit are sized to hold the remainder of the predetermined amount of blood. The neck portion of the delivery conduit is made of a material that can be sealed such that the container can be divided into a number of separate, independent compartments, and the neck portion of the delivery conduit is another use of the container. Summary of the Invention

[0010] The present invention, in its most general sense, relates to an apparatus for separating and recovering blood components by means of two conical chambers connected via their bases, the first chamber having a conduit for supplying a fluid to be processed and a device for recovering at least one component, characterized in that the second chamber comprises an elastically deformable flexible membrane and a bag, the flexible membrane extending to the conduit for laterally separating the space communicating with the second chamber, and the volume of the bag varying according to the deformation of the membrane.

[0011] Advantageously, the recovery device includes at least one float having a defined density and positioned in a channel formed in the junction between the conical chambers.

[0012] According to a variant, the recovery device includes a three-way valve positioned in a channel formed in the junction between the conical chambers, the channel including a collection nozzle.

[0013] The present invention also relates to an apparatus for separating and recovering blood components, the apparatus comprising a centrifuge having a plate, characterized in that the plate has at least one recess for use with the aforementioned apparatus.

[0014] Preferably, the plate includes at least one recess for the aforementioned device and includes an image sensor whose field of view corresponds to a passage area during rotation of a segment communicating between the chamber and at least a portion adjacent to each of the two chambers, and the plate includes a device for controlling the position of a valve of the device disposed on the plate.

[0015] Advantageously, the device also includes a computer that controls the controller based on the development of the image acquired by the image sensor. Attached Figure Description

[0016] The present invention will be more clearly understood by reading the following description, which refers to the accompanying drawings and relates to non-limiting embodiments of the invention, in which:

[0017] Figure 1 This is a longitudinal cross-sectional view of the device according to the present invention;

[0018] Figure 2 This is a longitudinal cross-sectional view of a variation of the device including a three-way valve according to the present invention;

[0019] Figure 3 This is a longitudinal cross-sectional view of the variant including the three-way valve at the blood filling step;

[0020] Figure 4 This is a longitudinal cross-sectional view of the variant including the three-way valve at the centrifugation step;

[0021] Figure 5 This is a longitudinal cross-sectional view of the variant including the three-way valve at the separation step;

[0022] Figure 6 It is a longitudinal cross-sectional view of the variant including the three-way valve at the deceleration step;

[0023] Figure 7 It is a longitudinal cross-sectional view of the variant including the three-way valve at the crank pin rotation step;

[0024] Figure 8 This is a longitudinal cross-sectional view of the variant including the three-way valve at the step of restoring deceleration;

[0025] Figure 9 It is a longitudinal cross-sectional view of the variant including the three-way valve at the step of re-establishing communication between the two cones of the device. Detailed Implementation

[0026] First Application Background of the Invention

[0027] The present invention will be described with reference to the following embodiments in the more specific context of cell therapy using blood components.

[0028] The invention described below relates to an apparatus intended for use in an automated system, preferably a single-use apparatus, which enables the processing and concentration of stem cells without loss or alteration of their function, particularly for the transplantation of hematopoietic progenitor stem cells.

[0029] The stem cell-rich components are isolated from peripheral blood taken from the circulatory system.

[0030] To isolate stem cells from the erythrocyte sedimentation rate (ESR) layer, density gradient products, such as those available under the names Ficoll and Percoll (trademarks), are used. The density gradient product is first introduced into the processing device, followed by whole blood. The components of the biofluid are then separated by centrifugation and collected by, for example, pipetting.

[0031] Physical description of the device

[0032] The centrifuge device according to the invention is formed from a molded portion made of plastic material, typically hourglass-shaped or biconical, and includes:

[0033] - An outer peripheral conical portion (10) includes a large base at its outer peripheral end that is closed by a cover (11), and the cover (11) includes a supply opening (12);

[0034] - A central conical portion (20), the central end of which is separated from the concave base (21) by an elastic membrane (22);

[0035] - Connecting sleeve (30) located between the base of the outer peripheral conical portion (10) and the base of the central conical portion (20).

[0036] The cover (11) of the outer conical portion (10) includes an outer peripheral rib (13) at the central portion of the cover. The outer peripheral rib (13) rests on an annular seal (14), which is received in an outer peripheral groove (15) provided at the outer peripheral edge of the outer conical portion (10). The outer peripheral edge (16) forms a hook-shaped element located in the chamber of a centrifuge (not shown).

[0037] The connecting sleeve (30) includes two channels (31, 32) that are open inside the central tapered portion (20).

[0038] The first channel (31) establishes a connection between the base of the outer conical portion (10) and the central conical portion (20). A first float (33) with a density of D1 is suitable.

[0039] The second channel (32) passes through the outer conical portion (10) and leads to the outside of the cover (11). The second channel encloses the first float (34) with density D2 and the second float (35) with density D3.

[0040] The membrane (22) is an elastically deformable biocompatible silicone membrane with approximately 450% elasticity and a thickness between 1 mm and 2 mm and a diameter of 50 mm. The membrane ensures a closed cover and an annular outer peripheral edge, which is sealed in an annular groove provided on the front portion of the central conical portion (20).

[0041] The outer conical portion (10) and the central conical portion (20) have an upper diameter of 50 mm, and the inner diameter of the connecting channel is between 1 mm and 1.5 mm.

[0042] Description of three-way valve variants

[0043] According to the description of the second variant of the centrifuge

[0044] The centrifuge device according to this variant is also formed from a molded part made of plastic material, typically hourglass-shaped or double-cone-shaped, and includes:

[0045] - An outer conical portion (10) is closed by a cap (11) including a supply opening (12);

[0046] - A central conical portion (20), the central end of which is separated from the concave base (21) by an elastic membrane (22);

[0047] - Connecting sleeve (30) located between the base of the outer peripheral conical portion (10) and the base of the central conical portion (20).

[0048] The connecting sleeve (30) includes a channel (36). The channel (36) establishes communication between the base of the outer peripheral conical portion (10) and the central conical portion (20). The channel (36) is provided with a crank pin controlling a three-way valve (37), which controls the following based on its position:

[0049] - Fluid flow between the two conical sections;

[0050] - The flow of fluid between the outer conical portion (10) and the collecting nozzle (38);

[0051] - Closing of the three-way valve (37).

[0052] The membrane (22) is an elastically deformable biocompatible silicone membrane with approximately 450% elasticity and a thickness between 1 mm and 2 mm and a diameter of 50 mm. The membrane (22) includes an annular outer peripheral edge (23) that engages between an annular groove (24) disposed on the front edge of the central conical portion (20) and a groove (25) formed in the concave base (21). The edge of the groove (25) includes ribs (26) that ensure that the central conical portion (20) is held in place by clamping.

[0053] Function Description

[0054] Figures 2 to 9 The diagram illustrates the centrifugation sequence for blood samples.

[0055] The blood sample is transferred from the sterile bag through a sterile tube to the peripheral conical portion (10), which connects the blood sample to the supply opening (12). Figure 3 ).

[0056] The device is then placed in a centrifuge, with the outer conical portion (10) positioned on the side of the central portion of the centrifuge and the central conical portion (20) positioned on the outer side. A three-way valve (37) is positioned in the pass position to allow blood contained in the outer conical portion (10) to flow towards the central conical portion (20) under centrifugal force. Figure 4 ).

[0057] Then stop the centrifuge to allow the different components of the blood to separate according to their density, thus gradually separating red blood cells, white blood cells, platelets, and plasma. Figure 5 ).

[0058] Then, the speed is gradually reduced so that the layer of interest rises directly below the separation region. Once the layer of interest has reached this region, a constant centrifugal velocity is switched: the layer is immobilized, and the elastic membrane balances the pressure ( Figure 6 ).

[0059] Then the three-way valve (37) is changed to the position used to establish communication with the collection nozzle (38) in order to allow platelet collection. Figure 7 ).

[0060] Then the three-way valve (37) is changed to the position used to establish communication with the collection nozzle (38) in order to allow platelet collection. Figure 7 ).

[0061] Return the centrifuge to a slow deceleration state—to the layer of interest ( Figure 8 It is emptied via a side collection nozzle (38).

[0062] Once the platelet layer recovers, an order is given to return to a constant rate. Figure 8 ).

[0063] Then return the crank pin controlling the three-way valve to the through position. Isolate the layer of interest ( Figure 9 ).

Claims

1. An apparatus for separating and recovering blood components by means of two conical chambers (10, 20), the conical chambers being connected via their bases, the two conical chambers comprising a first conical chamber (10) and a second conical chamber (20), the first conical chamber (10) having a supply opening for supplying a fluid to be processed and means for recovering at least one component, Its features are, The second conical chamber (20) includes an elastically deformable flexible membrane (22) and a concave base (21), the flexible membrane (22) laterally separating the second conical chamber (20) into a space for containing blood components and the concave base (21), the space leading to a channel for communicating with the second conical chamber (20), the device being configured such that blood components contained in the first conical chamber (10) can flow to the space of the second conical chamber under centrifugal force, and the volume of the concave base (21) varies according to the deformation of the flexible membrane, the flexible membrane being configured to balance the pressure of the space during centrifugation, thereby fixing the layers formed by different components of blood in the space.

2. The apparatus according to claim 1, characterized in that, The device for recovering at least one component includes at least one float having a defined density and positioned in the channel formed in the junction between the first conical chamber (10) and the second conical chamber (20).

3. The apparatus according to claim 1, characterized in that, The device for recovering at least one component includes a three-way valve (37) positioned in a channel formed in the junction between the first conical chamber and the second conical chamber, the channel including a collection nozzle.

4. An apparatus for separating and recovering blood components, said apparatus comprising a centrifuge having plates, characterized in that, The plate has at least one recess for use with the device according to any one of claims 1 to 3.

5. The apparatus for separating and recovering blood components according to claim 4, characterized in that, The plate includes at least one recess for use with the device according to claim 3, and the plate includes an image sensor whose field of view corresponds to a passage area during rotation of the crank pin in the channel to selectively establish communication between the first conical chamber (10) and the second conical chamber (20), and the plate includes a controller for controlling the position of the three-way valve of the device disposed on the plate.

6. The apparatus for separating and recovering blood components according to claim 5, characterized in that, The device also includes a computer that controls the controller based on the development of the image acquired by the image sensor.

Images