TW201742641A - Blood separation system - Google Patents

Abstract

A separation system comprising a blood separation device containing a controller, a red-blood-cell (RBC) detection module, and a pump driver; and a cartridge configured to be reversibly attached to the blood separation device, the cartridge including: (i) a first container configured to contain a blood sample, wherein the first container includes a first opening; (ii) a tubing in fluid flow communication with the first opening; (iii) a pump in fluid flow communication with the tubing, wherein the pump is configured to move a fluid from the first container to the pump and to contain a fluid; and (iv) a check valve mounted to the tubing, wherein the check valve is configured to allow a fluid to flow from the first container to the pump and to not allow a fluid to flow from the pump to the first container.

Description

Blood separation system

The disclosure of this case relates to the blood separation system.

BACKGROUND OF THE INVENTION Stem cells have the ability to self-renew to generate more stem cells and can also become almost any type of specialized cells. Stem cell research is useful for understanding human development and is one of the most attractive areas of contemporary biology. Therefore, stem cells provide exciting hope for future medical science.

SUMMARY OF THE INVENTION In one aspect, described herein is a separation system comprising: a controller, a red blood cell (RBC) detection module, and a pump driver (pump driver) a blood separation device; and a cartridge configured to be reversibly attached to the blood separation device. The crucible comprises: (i) a first container configured to contain a blood sample, wherein the first container comprises a first opening; (ii) a pipe ( Tubing) in fluid flow communication with the first opening; (iii) a pump in fluid flow communication with the pipe, wherein the pump is configured to draw a fluid from the first container Moving to the pump and for containing a fluid; and (iv) a check valve mounted to the pipe, wherein the check valve is configured to allow a fluid to flow from the first container to The pump also does not allow a fluid to flow from the pump to the first container. In a specific example, the RBC detection module is configured to (i) detect a position in a pipe at a detection point between the first opening and the pump. The red blood cell, and (ii) when the red blood cell is detected within the pipe, produces a first signal to be transmitted to the controller. The RBC detection module can be configured to emit a light to the detection point and detect reflection, scattering, and light that clearly indicates that the red blood cell is present in the fluid. Absorption or fluorescence.

The controller can be configured to receive the first signal from the RBC detection module and to generate a second signal to be transmitted to the pump driver after receiving the first signal. The pump driver can be configured to receive the second signal and to drive the pump to stop moving a fluid from the first container to the pump after receiving the second signal. The blood separation device can further include a temperature sensor configured to detect a temperature of a fluid located in the first container and to generate a temperature to be transmitted when the temperature is detected The third signal of the controller. The blood separation device can further include a cooling driver coupled to the controller, and wherein the controller is configured to generate and transmit a fourth signal to the third signal after receiving the third signal Cool the drive. The cooling driver can be configured to receive the fourth signal and, after receiving the fourth signal, drive a cooling module to modulate the temperature of a fluid located within the first container. The separation device can further include a holder configured to press the first container against the cooling module. The separation device can also include a heatsink adapted to transfer heat from the cooling module to the ambient of the blood separation device. In one embodiment, the separation device is configured to communicate with a central control unit via wireless communication. Furthermore, the separation device can be configured to reversibly lock the crucible in the separation device.

In one embodiment, the crucible further includes a second container in fluid flow communication with the pump, wherein the pump is adapted to move a fluid from the pump to the second container. The pump driver can be configured to, after receiving a driving signal from the controller, drive the pump to move a fluid from the pump to the second container. The file is adapted to allow a blood sample located within the first container to be separated into an upper layer and a gravity by gravity when fitted within the blood separation device Lower layer. Both the pump and the second container can be configured to be reversibly attached to the file.

In another aspect, a method for isolating a blood sample is described herein. For example, the method can include the use of a separation system as described in the disclosure of the present disclosure. In one embodiment, the method comprises: providing a mash comprising: (i) a first container comprising a blood sample mixed with a divalent cation chelating anticoagulant, Wherein the first container comprises a first opening; (ii) a pipe in fluid flow communication with the first opening; (iii) a pump in fluid flow communication with the pipe, wherein the pump is reversibly attached to The raft is also configured to move a fluid from the first container to the pump and to hold a fluid; and (iv) a check valve mounted to the pipe, wherein the check valve Configuring to allow a fluid to flow from the first container to the pump and not allowing a fluid to flow from the pump to the first container, wherein the file is configured to be reversibly attached to a blood separation device; Attaching the file to the blood separation device such that the first container is in an upright position and the first opening is in the top of the container; within the first container Blood sample is cooled to 2 ° C and 12 ° C; maintaining the blood sample at 2 ° C and 12 ° C for 6 to 72 hours, whereby the blood sample is separated by gravity into an upper layer and a lower layer; and a portion of the upper layer (a portion of the upper Pumping a red blood cell from the first container through the first opening and the pipe to the pump until the position in the pipe is at a detection point between the first opening and the pump It is detected that the pump maintains the portion of the upper layer. In one embodiment, the crucible further includes a second container in fluid flow communication with the pump, and wherein the method further includes a portion of the upper portion of the portion of the pump The upper layer is a step of pumping to the second container, wherein the second container holds the part of the portion of the upper layer.

Details of one or more specific examples are set forth in the accompanying drawings and the detailed description below. Other features, objects, and advantages of the specific embodiments will be apparent from the description and appended claims.

DETAILED DESCRIPTION The present disclosure sets forth a separation system that is adapted to separate a blood sample derived from a body, such as a patient. The system includes a blood separation device capable of communicating with a central control unit (eg, a desktop or hand-held computer) via a wireless or wired communications network And a crucible configured to be positioned within the blood separation device.

The size (Z) of a cell such as an adult stem cell mentioned in all of the following paragraphs of the disclosure of the present disclosure can be described or defined as, but not limited to: (1) in the field of cell biology (field of cell biology) or field of stem cells, the conventional definition of the size or representative length of a cell, (2) the diameter of a cell, especially When the cell is substantially spherical, (3) the length of a major axis of a cell, especially when the cell is substantially ellipsoidal, (4) when a cell When the shape has an approximate shape of a square, the width of the cell, (5) when the shape of a cell has an approximate shape of a rectangle, The length of the cell, or (6) the cross-sectional or transverse dimension of a cell. The size (Z), which is diameter, length, width or maximum cross-sectional or lateral dimension, can be determined or measured, but is not limited to, for example, using the cells from an optical microscope or a scanning electron. An image obtained by an electron microscope of a scanning electron microscope (SEM), or data obtained from a flow cytometer using the cells [eg, two-dimensional points, contours, or densities) Two-dimensional dot, contour or density plot]. The image obtained by the cell from an optical microscope or an electron microscope may be a two-dimensional (2D) cross section or a three-dimensional (3D) structure of the cell. As an example, the size (Z) of the cell may be, for example, by measuring the maximum cross-sectional or lateral dimension of the cell in a 2D cross-sectional image obtained from an optical microscope or an electron microscope (eg, SEM). And was obtained.

Adult stem cells [also known as adult stem cells] can be found in organs or tissues such as bone marrow, fat or peripheral blood and have the same basics of all stem cells. Same basic characteristics. An adult stem cell is an unspecialized or undifferentiated cell that can differentiate into a specialized cell type. In the disclosure of the present disclosure, adult stem cells are not embryonic stem cells; in other words, the adult stem cells are not derived, sourced, or harvested from embryos or fetal tissues. (fetal tissue).

There are various types of adult stem cells, including totipotent stem cells, pluripotent stem cells, multipotent stem cells, and progenitor stem cells [also known as single energy Unipotent stem cells]. The blastomere-like stem cells (BLSCs) are pluripotent stem cells or omnipotent stem cells. Very small embryonic-like stem cells (VSELs) are universal adult stem cells. SB-1 cells and SB-2 cells are univalent or multipotent adult stem cells.

Referring to Figure 1, a blood separation system includes a blood separation device 1 adapted to communicate with a central control unit (e.g., a desktop or handheld computer) via a wireless or wired communication network, and a configured The disposable crucible 2 to be positioned within the blood separation device 1 is formed. The blood separation system is capable of separating a blood sample, such as a peripheral blood sample from a body. The individual, for example, is a human [eg, a child, a teenager, an adult, or an elderly person] or an animal (eg, a mammal). The blood sample contains several cells, including a small-cell portion and a large-cell portion. The small-cell portion of the blood sample contains between 1 micrometer and 6 micrometers (and more preferably in size) as defined by the size (Z) of the cells described above. Small cells between 2 microns and 6 microns). The small-cell fraction contains platelets (which may be less than 6 microns in size) and small stem cells. The small stem cells [each of which have one or more nuclei or nuclei] contain small adult stem cells (which may be less than or equal to 6 microns in size), such as CD349(+) adult stem cells. , Lgr5 (+) adult stem cells, CD66e (+) adult stem cells (ie, BLSCs), and VSELs [eg, CD133 (+) adult stem cells and CD34 (+) adult stem cells]. The large-cell portion of the blood sample contains large cells larger than 6 microns in size, as defined by the size (Z) of the cells described above, such as large adult stem cells having a size system greater than 6 microns and Lineage cells containing red blood cells and white blood cells. After being processed, the blood sample is separated into two or more separate layers comprising an upper layer [e.g., a supernatant liquid] and a lower layer. The upper layer of the blood sample contains the small-cell portion, and the lower layer of the blood sample contains the large-cell portion. The separation system is adapted to separate the upper layer of the blood sample from the lower layer of the blood sample.

The disposable 匣2, for example, may be a pre-sterilized single-use cartridge, which includes a closed system for blood by gravity Separation is carried out inside. Referring to Figure 1, the disposable crucible 2 contains: (1) a container adapted to contain the blood sample, such as a blood container 23 [e.g., a 50 ml, 75 ml, 100 ml, 150 Mol, 200 ml, 250 ml, 300 ml, 350 ml, 400 ml, 450 ml or 500 ml blood bag], (2) a tube, for example a series of fluid flow connections with the blood container 23 Or system tubes 26a, 26b and 26c, (3) a check valve, such as a check valve 27 mounted to the pump tube in fluid flow communication (e.g., disposed between the tubes 26a and 26b) (4) a reversibly attached first container , such as a tube 26b and 26c or a syringe 24, and (5) a reversibly attached second container , such as a The tube 26c is in the form of a syringe 25 in fluid flow communication.

The disposable cartridge 2 is configured to be reversibly positioned or inserted into the blood separation device 1 such that when the disposable cartridge 2 is positioned or inserted within the blood separation device 1, the blood The container 23 is in the upright position and the two openings 23a and 23b are tied to the top of the blood container 23. A Luer lock connector 29 for blood collection is coupled to the opening 23a of the blood container 23 such that the blood sample flows from the individual to the blood via the Luer lock fitting 29 Inside the container 23. The tube 26a is connected to the opening 23b of the blood container 23 such that the upper layer of the blood sample flows from the blood container 23 into the tube 26a via the opening 23b of the blood container 23.

The check valve 27 (connecting the tube 26a and the tube 26b) may allow a fluid (eg, the upper layer of the blood sample) to be in only a first direction [ie, from the blood container 23 to the first syringe 24) a mechanical direction and a one-way directional valve that blocks the fluid in a second direction (ie, from the first syringe 24 to the blood container 23) The reverse) comes to flow. The fluid flow in the first direction opens the check valve 27, and the backflow in the second direction forces the check valve 27 to close. Accordingly, the check valve 27 (mounted between the tubes 26a and 26b) is adapted to sequentially pass the blood sample from the blood container 23 via the tube 26a in the first direction. The return valve 27 and the tube 26b flow to the first syringe 24 and prevent the blood sample from flowing from the first syringe 24 to the blood container 23 in the second direction.

The first syringe 24 is adapted to collect and contain an upper layer of the blood sample (containing the small-cell portion of the blood sample) located in the blood container 23 after gravity separation. The first syringe 24 is a pump comprising a first barrel 24a [e.g., a cylindrical tube], a first plunger located within the first barrel 24a. 24b and a first opening 24c. The second syringe 25 is adapted to contain an upper layer of the blood sample flowing from the first syringe 24 via the tube 26c. The second syringe 25 includes, for example, a second barrel 25a (e.g., a cylindrical tube), a second plunger 25b positioned within the second barrel 25a, and a second Second opening 25c. The first syringe 24 can be adapted to contain 10% to 200% more than the second syringe 25 (eg, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90) %, 100%, 150% or 200%) of the fluid. For example, the first syringe 24 can hold 60 ml of fluid while the second syringe 25 can hold 30 ml of fluid.

The first plunger 24b is configured to be driven by a pump driver 5 of the blood separation device 1 to be along an axial direction of the first barrel 24a of the first syringe 24 relative to the The first cylinder 24a moves. For example, when the first plunger 24b is driven by the pump driver 5 and is pulled along the axial direction relative to the first barrel 24a, the upper layer of the blood sample is sequentially ordered from the blood container 23. The ground flows into the first cylinder 24a via the tube 26a, the check valve 27, the tube 26b, and the first opening 24c. A disposable stop 28a is adapted to prevent the first plunger 24b from coming out of the first barrel 24a in the event of a malfunction of the pump driver 5. When the first plunger 24b of the first syringe 24 is driven by the pump driver 5 and is pushed along the axial direction relative to the first barrel 24a, a portion of the first barrel 24a is located The upper layer of the blood sample flows into the second barrel 25a of the second syringe 25 via the tube 26c and the second opening 25c. One of the disposal plates 2b of the disposable crucible 2b is adapted to prevent the second plunger 25b from coming out of the second cylinder 25a in the event of a failure of the pump driver 5. In summary, the first plunger 24b can be pulled and pushed within the first barrel 24a by the pump driver 5 of the blood separation device 1, allowing the first syringe 24 to be sucked in and discharged through the opening 24c. A liquid (eg, the upper layer of the blood sample).

The disposable crucible 2 further includes a divalent cation chelating-based anticoagulant that is preloaded within the blood container 23 for mixing with the blood sample. For example, the anticoagulant based on divalent cation chelation is ethylenediaminetetraacetic acid (EDTA), citrate or other calcium chelate anticoagulant (calcium- Chelating anticoagulant). In one embodiment, the blood container 23 contains an amount of the anticoagulant based on the divalent cation chelation, and after the anticoagulant is mixed with the blood sample, the blood sample is in each milliliter. The blood sample contains 1.5 mg or more (eg 1.5 mg to 2 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg or 2 mg) of anticoagulant. In one embodiment, as shown in Figure 2, the disposable cassette 2 further includes a filter 31 that is mounted to the tube 26a. The filter 31 is adapted to filter leukocytes in the blood sample.

Referring to Figure 1, the blood separation device 1 (which is adapted for use with the disposable cartridge 2 to separate a blood sample located in the blood container 23 of the disposable cartridge 2) includes a controller 3 coupled to The RBC detection module 4 of the controller 3 and the pump driver 5 (e.g., a pump or actuator) coupled to the controller 3. The RBC detection module 4 is adapted to detect red blood cells in the blood sample at a detection point 30 when the blood sample flows through the tube 26a, to generate a to be transmitted to the control The detection signal of the device 3. The RBC detection module 4 includes, for example, a light emitting device adapted to emit a light to the tube 26a located at the detecting point 30, and an adapted light emitting device. A sensor that reflects, reflects, scatters, absorbs, or fluoresces the red blood cells present in the blood sample of the tube 26a flowing through the detection point 30 is detected. The RBC detection module 4 can be placed such that a flow path between the opening 23a and the detection point 30 has a distance greater than the detection point 30 and the first The flow passage between the openings 24c is low (for example, 1 to 95%, 3%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45) %, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%). The RBC detection module 4 is adapted to perform a red blood cell detection when the tube 26a located at the detection point 30 is clear. The RBC detection module 4 is adapted to tolerate the presence of bubbles, "flakes" [stray red blood cells] and between the upper layer of the blood sample and the lower layer of the blood sample The spread of the line prevents the premature generation of the detection signal that will end the pumping before the upper layer completely transfers from the blood container 23 to the first syringe 24.

The RBC detection module 4 can be adapted to perform the processing procedure shown in FIG. The light emitting device may include a first light-emitting diode (LED) emitting blue light having a wavelength of 461 nm, and a second green light emitting light emitting green light having a wavelength of 565 nm. The sensor can include one or more photodiodes (eg, photo photodiodes) that are adapted to detect light emitted from each of the LEDs. The RBC detection module 4 can be adapted to perform a calibration sequence prior to being positioned in the blood separation device. After receiving a calibration request (eg, from the central control unit or the blood separation device), there are light levels for the green and the blue channels. The LEDs are measured with the LEDs on and the LEDs off. The currents associated with the individual LEDs are independently set, requiring only one light measurement at a time, alternating rapidly between the blue and green LEDs. These calibration measurements are performed when the crucible is not located within the blood separation device. The calibration process sets maximum bright and maximum dark levels. These levels, as well as the currents drawn by the LEDs, can be checked to determine if they are within the expected expected bands. The absorption levels are then scaled between the two levels for both the green and blue channels. See Figure 4.

Referring to FIG. 3, the RBC detection module has three output signal pins, that is, status 1 (Status 1), status 2 (Status 2), and fault (Fault). The detector also has an input signal pin, which is a calibration requirement. If the detector passes the correction, the output pin will indicate a "No Tube" condition, such as the absence of the defect within the blood separation device. This "tubeless" condition occurs when both the green and blue light channels detect near maximum brightness, such as a minimum amount of light absorption. The next absorption level occurs in a "Top Layer" condition, such as when an upper layer from a portion of the blood container in the cassette flows through the detection point. This green light channel is more suitable for measuring this condition. If the green absorption level is below a certain level (see Figure 4), the output pins will indicate a "top" condition. The next absorption level occurs in an "Empty Tube" condition. This is a tube with nothing inside, or if a bubble flows through the detection point. This blue light channel is more suitable for measuring this condition. If the blue absorption level is below a certain level (see Figure 4), the output pins will indicate an "empty tube" condition. The last absorption level occurs in a "Blood" condition, such as when a red blood cell flows through a detection point. This blue light channel is more suitable for measuring this condition. If the blue light absorption level exceeds a certain level (see Figure 4), the output pins will indicate a "blood" condition.

Referring to FIG. 1, the controller 3 is, for example, a microcontroller, which is a single integrated circuit chip including a processor core and a memory ( Memory) and programmable input/output peripherals. The controller 3 is adapted to receive a detection signal from the RBC detection module 4 and generate a drive signal to be transmitted to the pump driver 5 after receiving the detection signal. The pump driver 5 is adapted to receive a drive signal from the controller 3, hold the first plunger 24b of the first syringe 24, and drive the first plunger 24b along the first barrel 24a along The axial movement of the first cylinder 24a. Therefore, based on the detection result or signal, the pump driver 5 is configured to drive a portion of the blood sample in the blood container 23 to sequentially flow through the detection point 30 located in the tube 26a, the check valve 27 The tube 26b and the opening 24c are inside the first barrel 24a.

The blood separation device 1 further includes a cooling module 6 [eg, a thermoelectric cooler, a vapor-compression cooler or a liquid cooler] adapted to be used Cooling a blood sample in the blood container 23, a temperature sensor 7 coupled to the controller 3, and a cooling driver 8 [eg, a thermoelectric driver] coupled to the controller 3 and the Both of the cooling modules 6 are adapted to drive the cooling module 6 based on a feedback from the temperature sensor 7. The temperature sensor 7 is adapted to sense a temperature associated with a blood sample in the blood container 23, such as an internal temperature of the disposable cassette 2, to produce a to be delivered A temperature sensor signal to the controller 3. The controller 3 is adapted to receive the temperature sensor signal and generate a cooling signal to be transmitted to the cooling driver 8 based on the temperature sensor signal. The cooling drive 8 is adapted to receive the cooling signal and to drive the cooling module 6 after receiving the cooling signal to cool the temperature associated with the blood sample in the blood container 23. In one embodiment, the controller 3, the cooling module 6, the temperature sensor 7, and the cooling driver 8 are adapted to cool and maintain a temperature associated with a blood sample in the blood container 23. To a temperature between 2 degrees Celsius (°C) and 12°C, such as 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C 2 to 7 ° C, 2 to 10 ° C, 4 to 8 ° C, 4 to 12 ° C, or 6 to 12 ° C. The support 11 of the blood separation device 1 can be used as a heat transfer mount that is adapted to remove heat from the blood container 23 to the cooling module 6. A passive heat exchanger 9 (e.g., a heat sink) of the blood separation device 1 is adapted to conduct heat from the cooling module 6 to the ambient of the blood separation device 1. A fan 10 of the blood separation device 1 is adapted to remove heat conducted to the passive heat exchanger 9 to the periphery of the blood separation device 1. In a specific example, the support member 11 is adapted to press the blood container 23 against the cooling module 6 to increase the contact surface between the blood container 23 and the cooling module 6 ).

The support member 11 of the blood separation device 1 can be adapted to hold the blood container 23 containing the blood sample to prevent it from falling down. The disposable crucible 2 is adapted to be locked within the blood separation device 1 by a locking mechanism 12 [e.g., an electric locking mechanism] of the blood separation device 1. The locking mechanism 12 is coupled to the controller 3 and is adapted to lock or unlock the disposable cassette 2 in response to a signal transmitted from the controller 3. In other words, the controller 3 is adapted to control the locking mechanism 12 to lock or release the disposable cassette 2. The locking mechanism 12 can be coupled to a disposable in-place switch or detector. Optionally, the seating switch or detector can be integrated into the locking mechanism 12. The position switch or detector is adapted to confirm that the disposable cassette 2 is properly positioned within the blood separation device 1.

Furthermore, the blood separation device 1 includes a wireless module 13 coupled to the controller 3, an identification (ID) module 14 coupled to the controller 3, coupled to A beeper 15 of the controller 3 is coupled to a start button 16 of the controller 3, a stop button coupled to the controller 3, a coupled to The status display s 18 of the controller 3, a liquid crystal display (LCD) 19 coupled to the controller 3, a test port coupled to the controller 3 A power entry module 21 and an AC-to-DC power supply module 22 coupled to the power entry module 21.

The wireless module 13 is adapted to allow wireless communication between the blood separation device 1 and the central control unit. A protocol used for the wireless communication may be a Wi-Fi communication protocol such as IEEE 802.11. The identification module 14 is adapted to have a unique identifier (i.e., an electronic product code) to be transmitted to the central control unit, which may be associated with a given system (given A single entity (e.g., the blood separation device 1) within a system (e.g., the separation system) is associated with a numeric or alphanumeric string. For example, the identification module 14 can be a radio-frequency identification (RFID) chip or an electronic tag, which can transmit the unique identification code via a radio wave. To the central control unit. The central control unit is capable of reading a unique identification code carried by the radio wave and identifying the blood separation device 1 via the unique identification code. Optionally, the identification module 14 can be an erasable programmable read-only memory (EPROM) chip that includes the unique identification code. The central control unit is capable of reading a unique identification code stored in the EPROM wafer and identifying the blood separation device 1 via the unique identification code.

The pager 15 can emit a sound to inform a user that an abnormal event or a problem has occurred within the blood separation device 1. The start button 16 is adapted to send a start signal to the controller 3 to initiate a process for separating blood samples in the blood container 23. The stop button 17 is adapted to send a stop signal to the controller 3 to abort or terminate the process for separating the blood sample. The status display 18, such as light emitting diodes (LEDs), can emit light to alert a user to a state of the blood separation device 1, such as a power-on and/or error state. . The liquid crystal display 19 [which is a flat-panel display or other electronic visual display using light-modulating properties of liquid crystals) is adapted to display the relevant Information used to isolate the processing of the blood sample. The test 埠 20, such as an RS-232 serial port, is adapted to be used during test and maintenance to download logs and to allow non-user diagnostics.

The power entry module 21, for example, can be an electrochemical component used in the blood separation device 1, integrating a power inlet with other components [such as a switch and a fuse Fuse holder]. The power entry module 21 is adapted to be connected to an external power source by a power supply cord and to provide an alternating current (AC) power input to the AC and DC. Power supply module 22. The AC/DC power supply module 22, such as an AC-to-DC voltage converter, is adapted to convert AC power from the power entry module 21 into DC power. (DC power) to supply all of these components 3-20.

The blood separation device 1 may further comprise a housing containing the above components 3-22. A space (e.g., a socket) located within the housing is configured to receive the disposable cassette to be detachably locked to the blood separation device 1 by the locking mechanism 12. 2. In the blood separation device 1, the controller 3 is adapted to control and coordinate all of the components 3-22. For example, the controller 3 can perform the following operations: (1) starting a processing procedure for separating the blood sample after receiving a start command from the central control unit or from the start button 16, (2) operating the locking mechanism 12 to lock the disposable cassette 2 or receive an instruction from the central control unit or the stop button 17 after receiving an instruction or signal from the central control unit or the activation button 16 The signal is then released from the disposable 匣2, (3) monitoring the circuit of the pump driver 5 to detect an excess current (if the first injector 24 is blocked) or no current (if the first The syringe 24 is not properly engaged with the pump driver 5, (4) completes the process even if it is dropped out in wireless or wired communication, and (5) receives a command from the central control unit or the stop button 17 The handler is interrupted after a stop command or signal. The controller 3 may include a battery backed up clock/timer to allow monitoring of the elapsed processing time of the processing routine for separating the blood sample, even via a power Loss of power. Optionally, the blood separation device 1 can include a keep-alive circuit adapted to receive periodic pulses from the controller 3 (at least once per second). If two or more pulses from the controller 3 are lost or not received by the living circuit, the power to the pump driver 5 and to the cooling module 6 is turned off.

In summary, the separation system comprises the blood separation device 1 (which is capable of communicating with a central control unit) as illustrated in Figure 1 and is configured to be positioned within the blood separation device 1 as shown in Figure 1 or This single use 匣2 illustrated in Figure 2. The central control unit is adapted to run software to allow a user to control one or more functions of the blood separation device 1 by wireless or wired communication. The blood separation device 1 can be in communication with the central control unit via its wireless module 13 or its Ethernet module. The blood separation device 1 comprises a reusable piece of equipment for all electronic, mechanical, optical and thermal control devices necessary for safely performing blood separation (electronic, mechanical, optical and thermal control devices). ) (for example, these components 3-22).

In another embodiment, a separation system is provided that includes at least two blood separation devices 1 (each as illustrated in Figure 1) and at least two are configured to be positioned individually A single use 匣2 within the blood separation device 1 (each as illustrated in Figure 1 or Figure 2). A central control unit is adapted to operate the software to allow a user to control the functionality of the one or more blood separation devices 1 by wireless or wired communication. Each of the blood separation devices 1 can be in communication with the central control unit via its wireless module 13 or its Ethernet module. The central control unit is connectable to a peripheral device capable of reading a serialized unique identifier integrated into each of the peers 2. For example, the peripheral device can be a barcode reader, and the serialized unique identification code can be a printed barcode. Optionally, the peripheral device can be a radio frequency identification (RFID) reader, and the serialized unique identification code can be an RFID code. Each of the serialized unique identification codes, for example, can be linked to information of another patient and can be read at one or more points during processing and treatment to prevent patient samples from being Accidental exchange. Each of the blood separation devices 1 comprises a piece of reusable device consisting of all of the electronic, mechanical, optical and thermal control devices (e.g., such components 3-22) necessary to perform blood separation safely. .

A method of separating a blood sample using the separation system is illustrated in FIG. Referring to Fig. 5, in a step S1, the blood sample is aseptically extracted from the blood of the individual (e.g., peripheral blood) to a position containing a divalent cation chelation within the disposable crucible 2. The blood sample 23 of the anticoagulant (e.g., EDTA or citrate) is mixed with the anticoagulant based on the divalent cation chelation. In a step S2, the disposable cassette 2 is inserted or positioned within the blood separation device 1, i.e., into a socket within the outer casing of the blood separation device 1, and is located in the disposable The blood container 23 with the blood sample within the crucible 2 is held by the support member 11. The disposable cartridge 2 is inserted or positioned within the blood separation device 1 such that the blood container 23 is in an upright position. Thus, each of the two openings 23a and 23b of the blood container 23 positioned within the disposable crucible 2 located in the blood separation device 1 is tied to the top of the blood container 23 and is at One is at a higher level on the gravity coordinate than the blood sample in the blood container 23 of the disposable cassette.

Next, after receiving an activation signal from the central control unit or the activation button 16 from the blood separation device 1, the blood separation device 1 sequentially performs the following steps S3-S5. In a step S3, the cooling module 6 of the blood separation device 1 is driven to cool a blood sample located within the blood container 23 of the disposable cartridge 2 and maintain it at a specific temperature for a duration. For a predetermined period of time, for example, 3 to 72 hours, 3 to 12 hours, 3 to 18 hours, 3 to 24 hours, 3 to 36 hours, 3 to 48 hours, 3 to 60 hours, 6 to 72 hours, 6 to 12 Hours, 6 to 18 hours, 6 to 24 hours, 6 to 36 hours, 6 to 48 hours, 6 to 60 hours, 12 to 72 hours, 12 to 18 hours, 12 to 24 hours, 12 to 36 hours, 12 to 48 Hours, 12 to 60 hours, 16 to 72 hours, 16 to 18 hours, 16 to 24 hours, 16 to 36 hours, 16 to 48 hours, 16 to 60 hours, 24 to 72 hours, 24 to 36 hours, 24 to 48 Hours, 24 to 60 hours, 36 to 72 hours, 36 to 48 hours, 36 to 60 hours, 48 to 72 hours, or 48 to 60 hours. The specific temperature may be 2 ° C and 12 ° C, such as 2 ° C, 3 ° C, 4 ° C, 5 ° C, 6 ° C, 7 ° C, 8 ° C, 9 ° C, 10 ° C, 11 ° C, 12 ° C, 2 to 7 ° C 2 to 10 ° C, 4 to 8 ° C, 4 to 12 ° C, or 6 to 12 ° C. After being stored at the specific temperature for a predetermined period of time, due to gravity, for example, only gravity, blood located in the blood container 23 of the disposable cartridge 2 positioned in the blood separation device 1 The sample is separated into two or more separate layers comprising an upper layer and a lower layer. The upper layer of the blood sample contains suspended small cells such as small stem-like cells (e.g., small adult stem cells) and platelets. Platelets located in the upper layer of the blood sample may be non-viable, for example. The lower layer of the blood sample contains deposited large cells such as large adult stem cells, red blood cells, and white blood cells. Thus, in this step S3, the blood separation device 1 functions by allowing gravity-driven sedimentation of large cells within the blood container 23 of the disposable cartridge 2, and the segment is scheduled to be The time is considered to be a sedimentation time.

In a step S4, the controller 3 transmits a first driving signal to the pump driver 5 after gravity separation. After receiving the first drive signal, the pump driver 5 drives the first plunger 24a to pull the first plunger 24a along the axial direction of the first barrel 24a with respect to the first barrel 24a. Then, the upper layer of the blood sample located in the blood container 23 is pumped out of the blood container 23, and sequentially passes through the tube 26a, the check valve 27, the tube 26b, and the first opening 24c to the first Inside a tube 24a. At the same time, when the blood sample flows through the tube 26a, the RBC detecting module 4 detects whether there is red blood cells in the blood sample at a detecting point 30, so as to generate a signal to be transmitted to the controller 3. The detection signal (which is transmitted to and received by the controller 3). When the RBC detection module 4 detects the light of the tube 26a indicating that the red blood cell reaches the detection point 30, it generates a detection signal and transmits the detection signal to the controller 3. After receiving the detection signal transmitted from the RBC detection module 4, the controller 3 generates a second driving signal and transmits the second driving signal to the pump driver 5. After receiving the second detection signal transmitted from the controller 3, the pump driver 5 stops pulling the first plunger 24a and draws the blood sample into the first barrel 24a. In a step S5, the controller 3 generates a third driving signal to command the pump driver 5 to push the first one along the axial direction of the first barrel 24a with respect to the first barrel 24a. The plunger 24a causes the upper layer of the blood sample collected in the first barrel 24a to flow into the second barrel 25a via the tube 26c. Once the second cartridge 25a is filled with the upper layer of the blood sample, the controller 3 generates a fourth driving signal to command the pump driver 5 to stop pushing the first plunger 24a and stop driving to be collected. The upper layer of the blood sample in the first cartridge 24a flows into the second cylinder 25a. The process for separating the blood sample is then completed and the blood separation device 1 signals the central control unit.

The blood separation device 1 continues to safely maintain the disposable crucible 2 below the specific temperature described above (e.g., between 2 ° C and 12 ° C) until the disposable crucible 2 is removed by a user. After the disposable cartridge 2 is removed from the blood separation device 1, the first syringe 24 and the second syringe 25 containing the upper layer of the blood sample can be detached from the disposable cartridge 2 for a person [eg, a clinician (clinician), a doctor (doctor) or a researcher (researcher)]. The upper layer of the blood sample located within the first syringe 24 or the second syringe 25 can be used as a therapeutic cell mixture or a solution containing stem cells for treating a disease or disorder, such as A cancer, arthritis [eg osteoarthritis, psoriatic arthritis, rheumatoid arthritis, ankylosing spondylitis] , tendonitis, tendon injury, or autoimmune disease or disorder [eg rheumatoid arthritis, joint adhesion spondylitis, or systemic lupus erythematosus (systemic lupus) Erythematosus)], used to treat joints, (muscle) tendon, (knee) articular cartilage, shoulder or spine, or for Bone- or joint-related treatment [eg, a dental implant treatment]. The upper layer of the blood sample located within the first syringe 24 or the second syringe 25 can be saved for other applications, such as medical analysis.

The blood separation device 1 can be operated by a medical technician, a nurse or a physician at a point of care according to a provided protocols. The separation system is adapted to prepare a solution containing stem cells (i.e., the upper layer of the blood sample) from a patient's whole blood at the point of care. The term "autologous cell" means a cell that is genetically an individual's own cell.

Without further elaboration, it is believed that one skilled in the art can <RTIgt; </ RTI> <RTIgt;

Other Specific Examples All of the features disclosed in the present specification can be combined in any combination. Each feature disclosed in this specification can be replaced by an alternative feature that provides the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is merely an exemplification of one of the equivalent or similar features of the generic series.

From the above technical description, those skilled in the art can easily determine the basic features of the specific examples described, and can make various changes to the specific examples without departing from the spirit and scope of the invention. The changes and modifications are made to adapt to a variety of different uses and conditions. Therefore, other specific examples also fall within the scope of the patent application of this case.

1‧‧‧ Blood separation device
2‧‧‧Disposable
3‧‧‧ Controller
4‧‧‧Red Blood Cell (RBC) Detection Module
5‧‧‧ pump driver
6‧‧‧Cooling module
7‧‧‧Temperature Sensor
8‧‧‧Cooling drive
9‧‧‧ Passive heat exchanger, heat sink
10‧‧‧fan
11‧‧‧Support
12‧‧‧Locking mechanism
13‧‧‧Wireless Module
14‧‧‧ Identification (ID) module
15‧‧‧Pager
16‧‧‧Start button
17‧‧‧stop button
18‧‧‧Status display
19‧‧‧LCD display
20‧‧‧Test埠
21‧‧‧Power Entry Module
22‧‧‧AC power supply module
23‧‧‧ blood container
23a, 23b‧‧‧ openings
24‧‧‧ pump, syringe, first syringe
24a‧‧‧ first tube
24b‧‧‧first plunger
24c‧‧‧first opening, opening
25‧‧‧Second syringe
25a‧‧‧second tube
25b‧‧‧second plunger
25c‧‧‧second opening
26a, 26b, 26c‧‧‧ tube
27‧‧‧ check valve
28a, 28b‧‧ ‧ baffle
29‧‧‧Ruhr locking joint, Luer lock
30‧‧‧Detection points
31‧‧‧Filter
S1, S2, S3, S4, S5‧‧ steps

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a schematic drawing showing a blood separation device according to one embodiment of the present disclosure having a disposable cartridge inserted therein.

Fig. 2 is a schematic view showing a blood separation device according to a specific example of the disclosure of the present invention having a disposable cartridge inserted therein.

3 is a flow chart illustrating a red blood cell detection process in accordance with one embodiment of the present disclosure.

4 is a graph showing light absorption threshold levels corresponding to different states in accordance with one embodiment of the present disclosure.

Figure 5 is a flow chart illustrating a method of separating a blood sample derived from a subject using a blood separation system in accordance with one embodiment of the present disclosure.

1‧‧‧ Blood separation device

2‧‧‧Disposable

3‧‧‧ Controller

4‧‧‧Red Blood Cell (RBC) Detection Module

5‧‧‧ pump driver

6‧‧‧Cooling module

7‧‧‧Temperature Sensor

8‧‧‧Cooling drive

9‧‧‧ Passive heat exchanger, heat sink

10‧‧‧fan

11‧‧‧Support

12‧‧‧Locking mechanism

13‧‧‧Wireless Module

14‧‧‧ Identification (ID) module

15‧‧‧Pager

16‧‧‧Start button

17‧‧‧stop button

18‧‧‧Status display

19‧‧‧LCD display

20‧‧‧Test埠

21‧‧‧Power Entry Module

22‧‧‧AC power supply module

23‧‧‧ blood container

23a, 23b‧‧‧ openings

24‧‧‧ pump, syringe, first syringe

24a‧‧‧ first tube

24b‧‧‧first plunger

24c‧‧‧first opening, opening

25‧‧‧Second syringe

25a‧‧‧second tube

25b‧‧‧second plunger

25c‧‧‧second opening

26a, 26b, 26c‧‧‧ tube

27‧‧‧ check valve

28a, 28b‧‧ ‧ baffle

29‧‧‧Ruhr locking joint, Luer lock

30‧‧‧Detection points

Claims (28)

A separation system comprising: a blood separation device including a controller, a red blood cell (RBC) detection module, and a pump driver; and a cassette configured to be reversibly attached to the blood separation device, The crucible comprises: (i) a first container configured to contain a blood sample, wherein the first container includes a first opening; (ii) a tube in fluid flow communication with the first opening; Iii) a pump in fluid flow communication with the pipe, wherein the pump is configured to move a fluid from the first container to the pump and to contain a fluid; and (iv) a pipe to be mounted to the pipe A check valve, wherein the check valve is configured to allow a fluid to flow from the first container to the pump and does not allow a fluid to flow from the pump to the first container. The system of claim 1, wherein the RBC detection module is configured to (i) detect red blood cells located in the pipe at a detection point between the first opening and the pump And (ii) when the red blood cell is detected to be within the pipe, generating a first signal to be transmitted to the controller. The system of claim 2, wherein the RBC detection module is configured to emit a light to the detection point and to detect reflection, scattering, absorption or fluorescence of the light that clearly indicates that the red blood cell is present in the fluid . The system of claim 2 or 3, wherein the controller is configured to receive the first signal from the RBC detection module and to generate a signal to be transmitted to the pump driver after receiving the first signal The second signal. The system of claim 4, wherein the pump driver is configured to receive the second signal and to drive the pump to stop moving a fluid from the first container to the pump after receiving the second signal. The system of any one of claims 1 to 5, wherein the blood separation device further comprises a temperature sensor configured to detect a temperature of a fluid located in the first container and when the temperature is detected A third signal to be transmitted to the controller is generated. The system of claim 6, wherein the blood separation device further comprises a cooling driver coupled to the controller, and wherein the controller is configured to generate and transmit a fourth signal to the cooling after receiving the third signal driver. The system of claim 7, wherein the cooling driver is configured to receive the fourth signal and to drive a cooling module to adjust a temperature of a fluid located within the first container after receiving the fourth signal. The system of any one of claims 1 to 8, wherein the separating device further comprises a support configured to press the first container against the cooling module. The system of any one of claims 1 to 9, wherein the separating device further comprises a heat sink adapted to transfer heat from the cooling module to the periphery of the blood separation device. The system of any one of claims 1 to 10, wherein the crucible further comprises a second container in fluid flow communication with the pump, wherein the pump is adapted to move a fluid from the pump to the second container . The system of claim 11, wherein the pump driver is configurable to drive the pump to move a fluid from the pump to the second container after receiving a drive signal from the controller. The system of any one of claims 1 to 12, wherein the sputum is adapted to allow a blood sample located within the first container to be separated by gravity when positioned within the blood separation device One upper layer and one lower layer. The system of any one of claims 1 to 13, wherein both the pump and the second container are configured to be reversibly attached to the crucible. The system of any one of claims 1 to 14, wherein the separating device is configured to communicate with a central control unit via wireless communication. The system of any one of claims 1 to 15, wherein the separating device is configured to reversibly lock the file in the separating device. A method for isolating a blood sample, comprising: providing a first container comprising: (i) a blood sample comprising a blood sample mixed with a divalent cation chelated anticoagulant, wherein the first a container comprising a first opening; (ii) a pipe in fluid flow communication with the first opening; (iii) a pump in fluid flow communication with the pipe, wherein the pump is reversibly attached to the port and Configuring to move a fluid from the first container to the pump and to hold a fluid; and (iv) a check valve mounted to the pipe, wherein the check valve is configured to allow a fluid flows from the first container to the pump and does not allow a fluid to flow from the pump to the first container, wherein the file is configured to be reversibly attached to a blood separation device; attaching the file to The blood separation device such that the first container is in an upright position and the first opening is in the top of the container; the blood sample in the first container is cooled to 2 ° C and 12 ° C; the blood sample is Maintained at 2 ° C and 12 ° C 6 to 72 hours, whereby the blood sample is separated into an upper layer and a lower layer by gravity; and a portion of the upper layer is pumped from the first container through the first opening and the pipe until the pump is in position The red blood cells in the tube are detected at a detection point between the first opening and the pump, wherein the pump holds the upper layer of the portion. The method of claim 19, wherein the crucible further comprises a second container in fluid flow communication with the pump, the method further comprising pumping a portion of the upper layer of the portion of the pump to the second container a step wherein the second container holds the portion of the upper layer of the portion. The method of claim 18, further comprising removing the cartridge from the blood separation device and removing the pump and the second container from the cartridge. The method of claim 17, wherein the blood separation device comprises a red blood cell (RBC) detection module and a pump driver, wherein the RBC detection module is configured to detect a position at the detection point The red blood cells within the tubing, and the pump driver are configured to drive the pump to move a fluid from the first container to the pump and to move a fluid from the pump to the second container. A blood separation device comprising: a controller, a cooling module configured to cool a blood sample, and a red blood cell (RBC) detection module configured to detect a bit at a detection point The red blood cells in the blood sample and a first signal are generated if the red blood cells are detected, and a pump driver is configured to drive a blood sample through the detection point. The blood separation device of claim 21, wherein the controller is configured to receive the first signal from the RBC detection module and to generate a signal to be transmitted to the pump driver after receiving the first signal The second signal. The blood separation device of claim 21, further comprising a support member configured to hold and press a container against the cooling module. The blood separation device of claim 21, wherein the RBC detection module is configured to emit a light to the detection point and to detect a reflection, scattering, absorption or reflection of the light that clearly indicates that the red blood cell is present in the fluid Fluorescent. The blood separation device of claim 21, further comprising a housing including the RBC detection module, the pump driver, and the cooling module, wherein a space within the housing is configured to accommodate a to be Removably locked to the helium of the blood separation device. A file configured to be positioned within a blood separation device as claimed in claim 21, comprising: a first container configured to contain a blood sample, wherein the file is configured to be positioned The blood separation device is such that the first container is held in an upright position within the blood separation device and has an opening at the top of the first container; a pipe in fluid flow communication with the opening; a pump in fluid flow communication with the pipe, wherein the pump is configured to contain a fluid; and a check valve mounted to the pipe, wherein the check valve is configured to allow a fluid from the The first container flows to the pump and does not allow a fluid to flow from the pump to the first container. The crucible of claim 26, further comprising a second container in fluid flow communication with the pump, wherein the second container is configured to contain a fluid flowing from the pump to the second container. The cartridge of claim 26, further comprising a filter mounted to the tubing, wherein the filter is configured to filter white blood cells from fluids that are first passed through the tubing.