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CN-121988466-A - Blood cell separation device for separating lymphocytes and/or stem cells from whole blood in an automated procedure

CN121988466ACN 121988466 ACN121988466 ACN 121988466ACN-121988466-A

Abstract

The present invention relates to a blood cell separation device for separating lymphocytes and/or stem cells from whole blood in an automated procedure, the device comprising a centrifugal chamber, a pump section, a detector device, an automated clamp and a processor device. Wherein the quality of the collected lymphocytes and/or stem cells is improved and the cell collection procedure is further automated by utilizing optical sensors comprised in the detector device for measuring turbidity and color in the claimed method and in the cell separator. The device of the present invention is particularly useful for harvesting lymphocytes and/or stem cell fractions from whole blood, wherein contamination of the harvested cell fraction with platelets, erythrocytes and granulocytes is reduced.

Inventors

  • Heck Mishurat
  • Michael brinkman

Assignees

  • 费森尤斯卡比德国有限公司

Dates

Publication Date
20260508
Application Date
20170817
Priority Date
20160906

Claims (14)

  1. 1. A blood cell separation device for separating lymphocytes and/or stem cells from whole blood in an automated procedure, the blood cell separation device comprising: a centrifugal chamber configured to receive a separation chamber for automatically separating whole blood in the separation chamber into a plasma fraction, a red blood cell fraction, and a buffy coat fraction comprising lymphocytes and/or stem cells; A pump portion configured to pump fluid through a tubing set mounted on the blood separation device and comprising the separation chamber, a collection container for lymphocytes and/or stem cells, and a plasma/buffy coat tube configured to contain a fluid flow exiting the separation chamber, wherein the buffy coat portion follows the plasma portion and the buffy coat portion is followed by the red blood cell portion; A detector device comprising an optical sensor configured to emit red light and green light through the fluid flow in the plasma/buffy coat tube and to receive at least a portion of the red light and green light after draining the fluid flow in the plasma/buffy coat tube, for determining the presence of lymphocytes and/or stem cells in the fluid flow in the plasma/buffy coat tube and for determining the presence of red blood cells in the fluid flow in the plasma/buffy coat tube; an automated clamp configured to be oriented in a first position for directing the fluid flow in the plasma/buffy coat in a direction away from the collection container and in a second position for directing the fluid flow in the plasma/buffy coat into the collection container, and A processor device configured to control the automatic clamp orientation in the second position to direct the fluid flow in the plasma/buffy coat tube into the collection vessel when the detector device measures a measurement indicative of the presence of lymphocytes and/or stem cells in the fluid flow in the plasma/buffy coat tube, and to control the automatic clamp orientation in the first position to direct the fluid flow in the plasma/buffy coat tube in a direction away from the collection vessel when the detector device determines that a measurement indicative of the presence of red blood cells in the fluid flow in the plasma/buffy coat tube is measured, The detector device monitors turbidity of the fluid flow in the plasma/buffy-line tube based at least in part on intensity of a portion of the red light received after the fluid flow in the plasma/buffy-line tube is expelled, The detector device monitors the color of the fluid flow in the plasma/buffy coat tube based at least in part on the intensity of the portion of the green light received after the fluid flow in the plasma/buffy coat tube is expelled, The processor means is configured to control the auto-clamp orientation in the second position to direct the flow of the plasma/buffy coat into the collection vessel and when the detector means determines that a change in intensity of a portion of the red light received after the flow of the fluid in the plasma/buffy coat is discharged reflects that turbidity of the flow of the fluid in the plasma/buffy coat has risen from a level indicating the absence of lymphocytes and/or stem cells in the flow of the fluid in the plasma/buffy coat to a level indicating the presence of lymphocytes and/or stem cells in the flow of the fluid in the plasma/buffy coat and that a change in intensity of a portion of the green light received after the flow of the fluid in the plasma/buffy coat is discharged reflects that a color of the flow of the fluid in the plasma/buffy coat indicates the presence of lymphocytes and/or stem cells in the plasma/buffy coat and at least a reduced platelet count The processor device is configured to control the automatic clamp orientation in the first position to direct the fluid flow in the plasma/buffy coat tube in a direction away from the collection container when the detector device determines that an intensity of a portion of the green light received after expelling the fluid flow in the plasma/buffy coat tube reflects that a predetermined threshold value of a color of the fluid flow in the plasma/buffy coat tube has been reached, wherein the predetermined threshold value of the color of the fluid flow in the plasma/buffy coat tube corresponds to a predetermined hematocrit value.
  2. 2. The blood cell separation device of claim 1 wherein the optical sensor includes a red LED for monitoring turbidity of the fluid flow in the plasma/buffy coat tube and a green LED for monitoring color of the fluid flow in the plasma/buffy coat tube.
  3. 3. The blood cell separation device according to any one of claims 1 to 2, wherein the detector device is further configured to monitor the green light and/or the red light to allow compensation for aging effects and temperature drift.
  4. 4. A blood cell separation device according to any one of claims 1 to 3 wherein the detector means is configured to alternately enable the red light and the green light.
  5. 5. The blood cell separation device according to any one of claims 1 to 4, wherein, The detector device is configured to send a signal to the processor device upon detection of the turbidity increase and the color change of the fluid flow in the plasma/buffy coat tube, and The processor device is configured to control the automatic clamp orientation in the second position to direct the fluid flow in the plasma/buffy coat into the collection container upon receipt of the signal from the detector device.
  6. 6. The blood cell separation device according to any one of claims 1 to 4, wherein, The detector means is configured to send a signal to the processor means upon determining that the color of the fluid flow in the plasma/buffy coat has reached the predetermined threshold, and The processor device is configured to control the automatic clamp orientation in the first position upon receipt of the signal from the detector device to direct the fluid flow in the plasma/buffy coat in a direction away from the collection container.
  7. 7. The blood cell separation device according to any one of claims 1 to 4 wherein the predetermined threshold value of the color of the fluid flow in the plasma/buffy coat tube corresponds to the red color of a fluid having a hematocrit value of up to 5%.
  8. 8. A blood cell separation device according to any one of claims 1 to 4 wherein the processor device is configured to control the automatic clip orientation in the first position after detecting the predetermined threshold of the color of the fluid flow in the plasma/buffy coat tube and after allowing an additional default volume of the fluid flow through the automatic clip.
  9. 9. The blood cell separation device according to claim 8, wherein, The processor device is further configured to control the centrifugal chamber to Separating the blood in the separation chamber into a red blood cell fraction and a plasma fraction, wherein the buffy coat fraction comprising the lymphocytes and/or stem cells is located between the red blood cell fraction and the plasma fraction, Accumulating the buffy coat fraction between the red blood cell fraction and the plasma fraction in the separation chamber while a fraction of the red blood cell fraction and the plasma fraction is removed from the separation chamber, wherein the fraction of the plasma fraction removed from the separation chamber flows through the plasma/buffy coat tube as part of the fluid stream, and Causing an overflow in which the buffy coat fraction and the subsequent red blood cell fraction exit the separation chamber and flow through the plasma/buffy coat tube after the fraction of the plasma fraction flows through the plasma/buffy coat tube as part of the fluid flow, and repeating the accumulating of the buffy coat fraction in the separation chamber, the overflowing into the plasma/buffy coat tube and the transferring of the buffy coat fraction comprising lymphocytes and/or stem cells into a collection container in a plurality of cycles, Allowing the additional default volume of the fluid flow through the autoclamp after detecting the predetermined threshold of the color of the fluid flow in the plasma/buffy coat tube in at least one first cycle of a first half of the plurality of cycles, before the collection of buffy coat fractions containing lymphocytes and/or stem cells is terminated for the at least one first cycle, wherein the first half of the plurality of cycles refers to the first five to ten cycles, and In at least one second cycle in a latter half of the plurality of cycles, after detecting the predetermined threshold value of the color of the fluid flow in the plasma/buffy coat tube, a reduced additional default volume of the fluid flow is allowed to pass through the autoclamp before collection of a buffy coat fraction containing lymphocytes and/or stem cells is terminated for the at least one second cycle, wherein the latter half of the plurality of cycles refers to a cycle after the first five to ten cycles.
  10. 10. The blood cell separation device according to any one of claims 1 to 4, wherein, The blood cell separation device is directly connected to a donor/patient, and the whole blood to be separated is obtained from the donor/patient via a needle and directly supplied to the blood cell separation device, and The plasma fraction and the red blood cell fraction are re-infused into the donor/patient via a second needle.
  11. 11. The blood cell separation device according to any one of claims 1 to 4, wherein, The blood cell separation device is not connected to the patient, The whole blood to be separated includes a blood sample that has been obtained from the patient, and The plasma fraction and the red blood cell fraction are not reinfused into the patient, but are collected in the collection container.
  12. 12. The blood cell separation device according to any one of claims 1 to 4, wherein the product collected in the collection container contains a large amount of lymphocytes and/or stem cells.
  13. 13. The blood cell separation device of any one of claims 1 to 4, wherein the product collected in the collection container has a reduced count of platelets, granulocytes and erythrocytes or is substantially free of platelets, granulocytes and erythrocytes.
  14. 14. The blood cell separation device of claim 13 wherein the product collected in the collection container has a hematocrit value of less than 10% when stem cells are collected as the product and less than 5% when lymphocytes are collected as the product.

Description

Blood cell separation device for separating lymphocytes and/or stem cells from whole blood in an automated procedure The present application is a divisional application of application number 201780054117.X (international application number PCT/EP 2017/070824), entitled "method for automatically collecting leukocytes from whole blood", having application date of 2017, 8, 17, priority date of 2016, 9, 6 (entering the national stage of china at 3, 4, 2019). Technical Field The present invention relates to a blood cell separation device for separating lymphocytes and/or stem cells from whole blood in an automated procedure and a method for separating stem cells and/or lymphocytes from whole blood in an automated blood separation system, wherein the quality of the collected product is improved and the cell collection procedure is further automated by using a detector comprising optical sensors for measuring the color and turbidity of the fluid flow in the claimed method and in the cell separation system, which can be used for performing the claimed method. Background The present invention relates to the field of blood separation. Apheresis is any procedure in which blood is withdrawn from a donor/patient and a portion (e.g., plasma, white blood cells, platelets, or red blood cells) is separated and retained, and the remainder is reinfused into the donor/patient. Various types of apheresis include erythrocyte apheresis, leukocyte apheresis, lymphocyte apheresis, stem cell apheresis, plasmapheresis, and thrombocytopenia (platelet apheresis). Apheresis is also used in therapy, where whole blood is separated into plasma, the major component of whole blood, and erythrocytes, and abnormal pathogenic components are removed by following plasma in a therapeutic plasmapheresis procedure, or abnormal erythrocytes are removed and replaced by healthy cells in a erythrocyte replacement procedure. In addition, in addition to plasmapheresis procedures and erythrocyte replacement procedures, there are therapeutic procedures to remove white blood cells (leukopenia) or to remove platelets (thrombocytopenia). Lymphocyte separation and stem cell collection are separation procedures to collect mononuclear cell components containing lymphocytes and/or stem cells (which are flushed from bone marrow into whole blood outside with a specific drug) from blood withdrawn from a donor/patient into whom the remainder of the blood is reinfused. Blood separation is typically performed using a blood cell separator. Such blood cell separators typically include a centrifuge unit that is used to separate blood components by density and size. Centrifugation methods can be divided into two basic categories, continuous Flow Centrifugation (CFC) and batch flow centrifugation. "continuous" in CFC processes means that blood is collected, spun and simultaneously returned. The main advantage of the method and system is the low in vitro volume of the procedure. This is particularly advantageous when a small population, children or patients with problems with the heart and circulatory system have to undergo an apheresis procedure. Intermittent flow centrifugation works in a cycle that includes the steps of drawing blood at a dose, spinning the blood, processing the blood, collecting one or more components, and refluxing the remaining components of the blood into the donor/patient. To prevent blood coagulation, anticoagulants are automatically mixed with the blood as it is pumped from the human body into the separation machine. The end product of the blood centrifugation process is settled red blood cells outside the separation chamber, buffy coat in the middle containing MNC (MNC is a mononuclear cell, i.e. lymphocytes and monocytes), stem cells and platelets, and plasma inside the separation chamber. In recent years, automated separation systems have been developed. Automatic clamps and sensors have been implemented in the device. Furthermore, cell separators with centrifugal chambers have been developed which are particularly suitable for the automatic separation of blood components. EP 0985453 discloses a corresponding centrifugal chamber for a cell separator, in particular for separating whole blood into its components. EP 1025872 relates to a tube set for a cell separator for separating blood into its components and to a cell separator using said tube set. EP 1025872 also relates to a method for performing blood separation. One known and commercially available cell separator is the com TEC cube cell separator (Fresenius). The COM TEC cube device utilizes continuous flow centrifugation to separate blood components. The blood components are collected outside the centrifuge. Furthermore, com. TEC is used with a disposable kit without sealing. Only a lower in vitro volume is required to operate the com TEC device. Donation and treatment may be performed using com TEC devices. For therapeutic applications, such as continuous photochemotherapy or Donor Lymphoc