MXPA99001874A - Systems and methods for collecting mononuclear cells employing control of packed red blood cell hematocrit - Google Patents

Abstract

Blood separation systems and methods employ a rotating chamber (12). The rotating chamber (12) includes an inlet region (38) where whole blood enters for centrifugal separation into packed red blood cells, a plasma constituent, and an interface carrying mononuclear cells between the packed red blood cells and the plasma constituent. The packed red blood cells in the rotating chamber (12) have a hematocrit value HPRBC. A controller operates in a first mode to convey whole blood into the inlet region (38) while removing packed red blood cells and the plasma constituent from the rotating chamber (12) and while maintaining the interface within the rotating chamber (12). The controller also operates in the first mode to maintain a set HPRBC by conveying packed red blood cells into the inlet region (38).

Description

SYSTEMS AND METHODS TO COLLECT CELLS MONONUCLEARS EMPLOYING HEMATOCRIT CONTROL OF RED BLOOD CELLS, PACKAGED Related Request This application is a continuation in part of the North American Patent Application, Copending, Serial No. 08 / 745,779, filed on November 8, 1996 and entitled "Blood Processing Systems and Methods for Collecting Mononuclear Cells," which is a division of US Patent Application Serial No. 08 / 472,750, filed June 7, 1995 (now US Patent No. 5,573,678).

Field of the Invention The invention relates to centrifugal processing systems and apparatus.

Background of the Invention Current blood collection arrangements routinely separate whole blood REF .: 29517 by centrifugation in its various therapeutic components, such as red blood cells, platelets, and plasma. Conventional blood processing systems and methods use durable centrifuge equipment in association with sterile processing chambers, for single use, typically made of plastic. The centrifuge equipment introduces whole blood into these chambers while spinning them to create a centrifugal field. Whole blood is separated within the rotating chamber under the influence of the centrifugal field in red blood cells, high density and plasma with platelets. An intermediate layer of leukocytes forms the interface between the red blood cells and the platelet-rich plasma. Mononuclear cells, (MNC) are present in the shell.

Brief Description of the Invention The invention provides systems and methods for separating mononuclear cells from whole blood. The systems and methods employ a rotatable camera. The camera has a region of inlet, where the whole blood is introduced for separation into packed red blood cells, a plasma constituent, and an interface having mononuclear cells between the red, packed blood cells and the plasma constituent. Red, packed blood cells have a hematocrit HPRBc value. The systems and methods transport the whole blood to the region of entry, while removing the packed red blood cells and the plasma constituent from the chamber, and also while maintaining the interface within the chamber. The systems and methods maintain a desired range of HPRB by transporting the red blood cells, packed to the region of entry with whole blood. Maintenance of HPRBc in a desired range maximizes the production of mononuclear cells and prevents contamination of the granulocytes. In a preferred embodiment, the systems and methods circulate red, packed blood cells that are removed from the chamber for transport to the entry region to control HPRBC • In a preferred embodiment, the systems and methods either maintain a desired range of the HWB whole blood hematocrit by circulating the plasma constituent in the whole blood. Maintaining HWB in a desired range maximizes the efficiencies of platelet separation in the chamber, leading to a purer production of mononuclear cells. Other features and advantages of the invention will become apparent in the revision of the following specification, drawings and appended claims.

Brief Description of the Drawings Figure 1 is a side sectional view of a blood centrifuge having a separation chamber incorporating the features of the invention; Figure 2 shows the cart element associated with the centrifuge shown in Figure 1, with an associated processing vessel wound around for use; Figure 3A is a perspective view of the centrifuge shown in Figure 1, with the cup and spool elements mounted for rotation in their access position; Figure 3B is a perspective view of the cup and spool elements in their mutual separation condition to allow securing of the processing vessel shown in Figure 2 around the spool member; Figure 4 is a plan view of the processing vessel shown in Figure 2; Figure 5 is a perspective view of a fluid circuit associated with the processing vessel, comprising cartridges mounted in association with pump stations in the centrifuge; Figure 6 is a schematic view of the fluid circuit shown in Figure 5; Figure 7 is a perspective view of the back side of a cartridge forming a part of the fluid circuit shown in Figure 6; Figure 8 is a perspective view of the front side of the cartridge shown in Figure 7; Figure 9 is a schematic view of the flow channels and valve stations formed within the cartridge shown in Figure 7; Figure 10 is a schematic view of a pump station proposed to receive a cartridge of the type shown in Figure 7; Figure 11 is a schematic view of the cartridge shown in Figure 9 mounted on the pump station shown in Figure 10; Figure 12 is a perspective view of a cartridge and a pump station forming a part of the fluid circuit shown in the Figure Figure 13 is a view of the upper part of a peristaltic pump forming a part of the fluid circuit shown in Figure 6, with the pump rotor in a retracted position; Figure 14 is a top view of a peristaltic pump forming a part of the fluid circuit shown in Figure 6, with the pump rotor in an extended position coupled to the pump tubing.

Figure 15 is a schematic top view of the separation chamber of the centrifuge shown in Figure 1, designed to show the radial contours of the high G and low G walls; Figures 16A and 16B somewhat schematically show a portion of the collection zone of platelet-rich plasma in the separation chamber, in which the surface of the high G wall forms a tapered wedge to contain and control the position of the interface. between red, packed blood cells and platelet-rich plasma; Figure 17 is a somewhat schematic view of the interior of the processing chamber, looking from the low G wall towards the high G wall in the region where the whole blood enters the processing chamber for the separation of red blood cells and platelet-rich plasma, and where the platelet-rich plasma is collected in the processing chamber; Figure 18 is a schematic view showing the established dynamic flow conditions defining, and "bordering" the MNCs within the blood separation chamber shown in Figure 17; Figure 19 is a schematic view of the process controller that configures the fluid circuit shown in Figure 6 to carry out a pre-set MNC collection processing; Figure 20 is a flowchart showing the cycles of variation and phases of the MNC collection processing that the controller shown in Figure 19 governs; Figure 21 is a schematic view showing the transport of blood and fluid components in the circuit shown in Figure 6 during the preliminary processing cycle of the procedure shown in Figure 20; Figure 22 is a schematic view showing the transport of blood components and fluids in the circuit shown in Figure 6 during the MNC accumulation phase of the processing shown in Figure 20; Figure 23 is a schematic view showing the transport of blood and fluid components in the circuit shown in Figure 6 during the PRBC collection phase of the processing shown in Figure 20; Figure 24A is a schematic view showing the transportation of blood components and fluids in the circuit shown in Figure 6 at the beginning of the MNC removal phase of the processing shown in Figure 20; Figure 24B is a schematic view showing the transport of blood and fluid components in the circuit shown in Figure 6 during the MNC removal phase of the processing shown in Figure 20; Figure 24C is a schematic view showing the transport of blood components and fluids in the circuit shown in Figure 6 at the end of the MNC removal phase of the processing shown in Figure 20; Figure 25 is a schematic view showing the transport of blood components and fluids in the circuit shown in Figure 6 during the flood phase of PRP - of the processing shown in Figure 20; Figure 26 is a schematic view showing the transport of blood components and fluids in the circuit shown in Figure 6 during the MNC suspension phase of the processing shown in Figure 20; Figure 27 is a schematic view showing the transport of blood components and fluids in the circuit shown in Figure 6 during the cleaning phase of the processing shown in Figure 20; Figure 28 is a schematic view of the optical sensor used in association with the circuit shown in Figure 6 to pursue and quantify the MNC region for collection; Figure 29 is an alternative embodiment of a fluid circuit suitable for collecting and collecting the MNCs; Figure 30 is a schematic view showing the transport of blood components and fluids in the circuit shown in Figure 29 during the PRBC collection phase of the processing shown in Figure 20; Figure 31 is a schematic view showing the transport of blood components and fluids in the circuit shown in Figure 29 during the MNC removal phase of the processing shown in Figure 20; The invention can also be incorporated in various forms without departing from its spirit or essential characteristics. The scope of the invention is defined in the appended claims, instead of the specific description that precedes them. All modalities that fall within the meaning and range of equivalence of the claims are therefore proposed to be encompassed by the claims.

Description of the Preferred Modalities The Centrifuge Figure 1 shows a blood centrifuge 10 having a blood processing chamber 12 suitable for collecting mononuclear cells (MNC) from whole blood. The limits of the chamber 12 are formed by a flexible processing vessel 14 carried within an annular gap 16 between a rotating reel element 18 and the cup element 20. In the illustrated and preferred embodiment, the processing vessel 14 takes the form of an elongated tube (see Figure 2), which is wound around the spool member 18 before use. Additional details of the centrifuge 10 are set forth in U.S. Patent No. 5,370,802, entitled "Enhanced Yield Platelet Systems and Methods," which is incorporated herein by reference. The cup and spool elements 18 and 20 are mounted on a yoke 22 between a vertical position, as shown in Figures 3A and 3B, and a suspended position, as shown in Figure 1. • When in the vertical position, the elements of cup and cart 18 and 20 are presented for access by the user. One mechanism allows the cup and spool members 18 and 20 to open, as shown in Figure 3B, so that the operator can roll the container 14 around the spool member 20, as shown in Figure 20. The spikes 150 in the spool member 20 couples the cuts in the container 14 to secure the container 14 in the spool member 20.

When closed, the spool and cup elements 18 and 20 can be mounted for rotation in the suspended position shown in Figure 1. In operation, the centrifuge 10 rotates the suspended cup and spool elements 18 and 20, about an axis 28, creating a centrifugal field within the processing chamber 12. Additional details of the mechanism for making a relative movement of the spool and cup elements 18 and 20 are described as described in US Pat. No. 5,360,542 entitled "Centrifugal With Separable Bowl and Spool Elements Providing Access to the Separation Chamber", which is incorporated herein by reference. The radial limits of the centrifugal field (see Figure 1) are formed by the inner wall 24 of the cup element 18 and the outer wall of the spool element 20. The inner wall 24 of the cup defines a high G wall. The outer wall 26 of the spool defines a low G wall.

I. The Processing Vessel In the illustrated embodiment (See Figure 4), a first peripheral seal 42 forms the outer edge of the container 14. A second inner seal 44 generally extends parallel to the rotational axis 28, dividing the container 14 into two compartments 38 and 40. In In use, whole blood is centrifuged in compartment 38. In use, compartment 40 carries a liquid, such as saline, to counterbalance compartment 38. In the embodiment shown in Figure 4, compartment 38 is larger than the compartment 40 by a volumetric ratio of approximately 1 to 1.2 Three ports 46, 48 and 50 communicate with the processing compartment 38, to transport the whole blood and its components. Two additional holes 52 and 54 communicate with the ballast compartment 40 to transport the counterbalancing fluid.

III. The Fluid Processing Circuit A fluid circuit 200 (see Figure 4) is coupled to the container 14. Figure 5 shows the general arrangement of the fluid circuit 200, as a function of a flexible pipe arrangement, liquid source and collection vessels, line pumps, and clamps, all of which will be described in detail later. Figure 6 shows the details of the fluid circuit 200 in schematic form. In the illustrated embodiment, the left, intermediate and right cartridges, respectively 23L, 23M and 23R, centralize much of the valve G and pumping functions of the fluid circuit 200. The left, intermediate, and right cartridges 23L, 23M and 23R are they couple with the left, middle, and right pump stations in the centrifuge 10, and are designated, respectively, PSL, PSM, and PSR.

A. The Cartridges Each cartridge 23L, 23M and 23R is constructed the same, so a description of a single 23L cartridge is applicable to all cartridges.

Figures 7 and 8 show the structural details of the cartridge 23L. The cartridge 23L comprises a molded plastic body 202. Flow channels 208 are integrally molded on the front side 204 of the body 202. A rigid panel 214 transports and seals the front side 204 of the body. Valve stations 210 are molded on the back side 206 of the body 202 of the cartridge. A flexible diaphragm 214 transports and seals the back side 206 of the body 202. Figure 9 schematically shows a representative arrangement of the flow channels 208 and the valve stations 210 for each cartridge. As shown, channels Cl through C6 intersect to form a star array, which diffuses from a central hub H. Channel C7 crosses channel C5; channel C8 crosses channel C6; channel C9 crosses channel C3; and the CÍO channel crosses the C2 channel. Of course, other channel patterns can be used. In this arrangement, the valve stations VS1, VS2, VS9 and VS10 are located in, respectively, the channels C2, C3, C5 and C6 immediately close to their common intersection in the hub H. The valve stations VS 3, VS4 , VS5, VS7 and VS8 are located in the outer extremities of channels C8, Cl, C2, C5, C4 and C3, respectively. Each cartridge 23L carries an upper flexible pipeline UL, which extends outside the cartridge 23L between the channels C7 and C6, and a lower pipeline circuit LL, which extends outside the cartridge between the channels C3 and CIO. In use, the UL and LL tube circuits couple the rotors of the peristaltic pump of the pumps in the associated pump station.

B Pumping Stations The pump stations PSL, PSM and PSR are, like the cartridges 23L, 23M and 23R, constructed identically, so that it is applicable to all a description of a PSL station. Figure 12 shows the structural details of the left pump station PSL. Figure 10 shows the left pump station PSL in a more schematic form. The PSL station includes two peristaltic pumps, for a total of six pumps in circuit 200, which are designated Pl to P6 (see Figure 6). The PSL station also includes an array of ten valve actuators (shown in Figure 10), for a total of 30 valve actuators in the circuit 200, which designate VA1 to VA30 (see Figure 6). In use (see Figure 11), the UL and LL circuits the 23L cartridge couple the Pl and P2 pumps of the PSL pump station. In a similar way (as shown in Figure 6); the UL and LL tube circuits of the intermediate cartridge 23M couple the pumps P3 and P4. The UL and LL tube circuits of the right cartridge 23L couple the pumps P5 and P6. As shown in Figure 11, the valve stations VSl to VS10 of the cartridge 23L are aligned with the valve actuators VI to VIO of the left pump station PSL. As shown in Figure 6, the valve stations of the intermediate and right cartridges 23M and 23R are likewise aligned with the valve actuators of the respective intermediate and right pump stations, PSM and PSR. The following Table 1 summarizes the operative association of the valve actuators VI to V30 of the pump station to the valve stations VSl to VS10 of the cartridge, shown in Figure 6.

Table 1: Alignment of the Valve Stations of the Cartridge to the Valve Actuators Table 1 (continued) The cartridges 23L, 23M and 23R are mounted at their respective pump stations PSL, PSM, PSR with their rear sides 206 facing down, so that the diaphragms 212 face towards and engage the valve actuators. Valve actuators Vn are solenoid-operated rods 215 (see Figure 12), which are biased to a position Vn valve. The valve actuators VN are molded to align with the valve stations VSn of the cartridge in the manner set forth in Table 1. When a given rod 215 is energized, the associated valve station of the cartridge is opened, allowing the passage of the liquid. When the rod 215 is not energized, the diaphragm 212 is moved to the associated valve station, blocking the passage of liquid through the associated valve station. In the illustrated embodiment, as shown in Figure 12, the pumps Pl to P6 in each pump station PSL, PSM and PSR include rotating peristaltic pump rotors 216. The rotors 216 can be moved between a retracted condition (as shown in FIG. 13), out of engagement with the respective tube circuit, and an operation condition (shown in FIG. 14), in which the rotors 216 engage the respective tube circuit against a pump stroke 218. The pumps Pl and P6 can be operated in this manner in three conditions: (i) a pump condition on, during which the pump rotors 216 rotate and are in their operating position to couple the pump pump tubing against pump stroke 218 (as shown in Figure 14). Therefore, rotating pump rotors 216 transport a fluid in a peristaltic manner through the pipe circuit. (ii) in a pump-off condition, open, during which the pump rotors 216 are not rotated and are in their retracted position, so as not to couple the pump tubing circuit (as shown in Figure 13). The pump-off, open condition thus allows the flow of fluid through the pump tube circuit in the absence of rotor rotation. (iii) in an off, closed pump condition, during which the pump rotors 216 are not rotated, and the pump rotors are in the operating condition. The stationary pump rotors 216 thus couple the pump tubing circuit, and serve as a clamp to block the flow of fluid through the pump tubing circuit. Of course, equivalent combinations of pump conditions can be achieved using peristaltic pump rotors that do not retract, by proper clamp placement and Pipe routes upstream and downstream of the pump rotors. The additional structural details of the cartridges 23L, 23M, 23R, the peristaltic pumps PI to P6, and the valve actuators VI to V30 are not essential to the invention. The details are described in U.S. Patent No. 5,427,509, entitled "Peristaltic Pump Tube Cassette with Angle Port Tube Connectors" (which is incorporated herein by reference).

C. The Fluid Flow Pipe The fluid circuit 200 further includes stretches of flexible plastic tubing, designated TI to T20 in Figure 6. The flexible tubing Ti to T20 couples the cartridges 23L, 23M and 23R to the processing vessel 14, to the external source and the bags or collection containers, and the blood donor / patient. The fluid flow function of the TI to T20 pipe in condition with the collection and collection of the MNCs will be described later. The following summarizes, from a point of view structural, the union of the TI pipe to T20, as shown in Figure 6: The TI pipeline extends from the donor / patient (via a conventional phlebotomy needle, not shown) through an external clamp C2 to the C4 channel of the external cartridge 23L. The pipe T2 extends from the tube TI through an external clamp C4 to the channel C5 of the intermediate cartridge 23M. The pipe T3 extends from an air detection chamber DI to the channel C9 of the left cartridge 23L. The pipe T4 extends from the drip chamber DI to the orifice 48 of the processing vessel 14. The pipe T5 extends from the orifice 50 of the processing vessel 14 to the channel C4 of the intermediate cartridge 23M. The pipe T6 extends from the channel C9 of the intermediate cartridge 23M to join the pipe T4 downstream of the chamber DI. The pipe T7 extends from the channel C8 of the right cartridge 23R to the channel C8 of the left cartridge 23L.

The pipe T8 extends from the channel Cl of the intermediate cartridge 23M to join the pipe T7. The pipe T9 extends from the channel C5 of the left cartridge 23L through an air detection chamber D2 and an external clamp C3 to the donor / patient (via a conventional phlebotomy needle, not shown). The UNC pipe extends from the orifice 46 of the processing vessel 14, through an optical sensor OS line to the C4 channel of the right cartridge 23R. The tubing Til extends from the channel C9 of the right cartridge 23R to the chamber DI. The pipe T12 extends from the channel C2 of the right cartridge 24R to the container proposed to receive the platelet poor plasma, designated PPP. A weight scale (not shown) receives the weight of the PPP container for the purpose of deriving the fluid volume changes. The pipe T13 extends from the channel Cl of the right cartridge 23R to a container proposed to receive the mononuclear cells, designated MNC. The pipe T14 extends from the cartridge C2 of the intermediate cartridge 23M to a proposed vessel for receiving red blood cells, packed, designated PRBC. A weight balance WS perceives the weight of the PRBC container for the purpose of deriving fluid volume changes. The line T15 extends from an anticoagulant container, designated ACD, to the channel C8 of the intermediate cartridge 23M. A weight scale (not shown) perceives the weight of the ACD vessel for the purpose of deriving fluid volume changes. Tubing T16 and T17 extend from a container of priming liquid, such as saline, designated PRIME, by bypassing all cartridges 23L, 23M and 23R, through an external clamp Cl, and by crossing, respectively, the tubing T9 (between the air detection chamber D2 and the clamp C3), and the TI pipe (upstream of the clamp C3). A weight scale (not shown) receives the weight of the PRIME container for the purpose of deriving fluid volume changes. The pipe T18 extends from the orifice 52 of the processing vessel 14 to the channel "C5 of the right cartridge 23R.

The pipe T19 extends from the orifice 54 of the processing vessel 14 to cross the pipe T18. The pipe T20 extends from the channel C2 of the left cartridge 23L to a container proposed to receive the waste priming fluid, designated WASTE. A weight scale (not shown) perceives the weight of the WASTE container for the purpose of deriving fluid volume changes. The portions of the pipe are joined at the navel 30 (see Figure 1). The belly button 30 provides communication for fluid flow between the interior of the processing vessel 14 within the centrifugal field and other stationary components of the circuit 200 located outside the centrifugal field. A non-rotating support 32 (zero omega) holds the upper position of the navel 30 in a non-rotating position above the suspended spool and cup elements 18 and 20.

A support 34 in the yoke 22 rotates the intermediate position of the navel 30 at a first speed (one omega) around the suspended spool and cup elements 18 and 20. Another support 36 rotates the lower end of the navel 30 to a second speed twice the speed one omega (speed two omega), to which the spool and cup elements, suspended 18 and 20 also rotate. This known relative rotation of the navel 30 maintains it without twisting, thus avoiding the need for rotating seals.

IV. Separation in the Blood Processing Chamber (A General View) Before explaining the details of the procedure by which the MNCs are collected using the container 14 and the fluid circuit 200, the fluid dynamics of the separation of the whole blood in the processing compartment 38 will generally be described first, with reference mainly to Figures 4 and 15 to 17. Referring first to Figure 4, whole blood with anticoagulant (WB) is removed from the donor / patient and transported to the processing compartment through port 48. The processing compartment Blood 38 includes an inner seal 60 and 66, which form a WB entry passage 72 leading to an entry region of WB 74.

As the WB follows a circumferential flow path in the compartment 38 around the rotational axis 28. The side walls of the container 14 extend to fit the profiles of the outer wall 26 (low G) of the spool member 18 and the inner wall 24 (high G) of the cup element 20. As shown in Figure 17, the WB is separated in the centrifugal field within compartment 38 of the blood processing in packed, red blood cells (PRBC, designated by number 96), which move towards wall 24 of high G, and platelet-rich plasma (PRP, designated pro number 98), which is displaced by the movement of PRBC 96 towards wall 26 of low G. An intermediate layer, called the The interframe (designated by the number 58) is formed between the PRBC 96 and the PRP 98. Referring again to Figure 4, the inner seal 60 also employs a PRP collection region 76 within the blood processing compartment 38. As further shown in Figure 17, the PRP collection region 76 is adjacent to the WB input region 74. The speed at which the PRBC 96 they settle towards the high-G wall 24 in response to the centrifugal force being larger in the WB inlet region 74 than elsewhere in the blood processing compartment 38. There is also relatively more plasma volume to travel towards the low G wall 26 in the WB input region 74. As a result, the relatively large radial plasma velocities towards low-G wall 26 occur in the WB input region 74. These large, radial velocities towards the low G wall 26 extract large numbers of platelets from the PRBC 96 in the PRP collection region 76, conveniently. As shown in Figure 4, the inner seal 66 also forms a closed curve 70 that defines a PRBC collection passage 78. A gradual bore 115 (see Figure 15) extends into the PRBC mass along the high G wall 24, creating a restricted passage 114 between it and the high G wall 24, and its radial, surface . The restricted passage 114 allows the PRBC 96 to be present along the high-G wall 24 to move beyond the barrier 115 to the PRBC collection region 50, for transportation through the PRBC collection passage 78 to the hole 50 of PRBC. Simultaneously, the barrier 115 is gradual what the passage of PRP 98 goes beyond it. As shown in Figures 15, 16A and 16B, the high-G wall 24 also projects toward the low-G wall 26 to form a tapered ramp 84 in the PRP collection region 76. The ramp 84 forms a narrow passage 90 along the low-G wall 26, along which the PRP layer 98 extends. The ramp 84 maintains the interface 58 and the PRBC 96 away from the collection orifice 46. of PRP, while allowing PRP 98 to reach office 46 of PRP collection. In the illustrated and preferred embodiment (see Figure 16A), the ramp 84 is oriented at an angle not parallel to less than 45 ° (and preferably about 30 °) with respect to the axis of the PRP-orifice 46. The angle to the overflow of the interface and the PRBC through the narrow passage 90. As shown in Figures 16A and 16B, the ramp 84 also exhibits the interface 26 for viewing through a side wall of the container 14 by a controller. 220 of entrecara, associated (see Figure 19). The interleaver controller 220 controls the relative flow rates of WB, PRBC, and PRP through their respective holes 48, 50 and 46. In this way, the controller 220 can maintain the interface 58 at pre-established locations on the ramp, either close to the narrow passage 90 (as shown in Figure 16A). Or away from the narrow passage 90 (as shown in Figure 16B). By controlling the position of the interface 58 on the ramp 84 relative to the narrow passage 90, the controller 220 can also control the platelet content of the plasma collected through the orifice 46. The concentration of platelets in the plasma increases with proximity at the 58th interface. By maintaining the 58-shell in a relatively low position on the ramp 84 (as shown in Figure 16B), the platelet-rich ramp is kept away from the hole 46, and the plasma conveyed through the orifice 46 has a relatively low platelet content. By keeping the collar 58 in a high position on the ramp 84 (as shown in Figure 16A), closer to the hole 46, the plasma carried by the hole 46 is rich in platelets. Alternatively, or in combination, the controller can control the location of the 58th interface by varying the speed at which introduces the WB into the blood processing compartment 38, the rate at which the PRBCs are transported from the blood processing compartment 134, both. Additional details of a preferred embodiment for the skinner controller are described in U.S. Patent No. 5,316,667, which is incorporated herein by reference. As shown in Figure 15, the radially opposed surfaces 88 and 104 form a region 108 restricted to flow along the high-G wall 24 of the WB inlet region 74. As also shown in Figure 17, region 108 restricts a flow of WB in WB input region 74 to a reduced passageway, thereby causing a more uniform perfusion of WB into blood processing compartment 38 as required. - length of the low G wall. This uniform perfusion of WB occurs adjacent to the PRP collection region 76 and in a plane that is approximately the same as the plane in which the preferred, controlled position of the 58 interface is. Once it is beyond the narrow region 108 of zone closure 104, the PRBC 96 is move quickly towards the wall of high G 24 high G in response to centrifugal force. The narrow region 108 places the WB in the entry region 74 at approximately the preferred, controlled height of the 58th interface. The WB placed in the entry region 74 below or above the controlled height of the 58th interface will immediately search the height of the interface and, in doing so, will oscillate around it, causing unwanted secondary fluxes and disturbances along the interface 58. By placing the WB in the entrance region 74 approximately at the level of the interface, the region 108 reduces the incidence of secondary flows and disturbances along the interphase 58. As shown in Figure 15, the low wall G 26 tapers away from the axis of rotation 28 toward the high wall 24. G in the direction of flow of WB, while the wall 24 of high G of surface retains a constant radius. The taper may be continuous (as shown in Figure 15) or may occur gradually. These contours along walls 24 and 26 of high G and low G produce a condition of circumferential, dynamic plasma flow, in general transverse to the centrifugal force field in the direction of the 76 PRP seclusion region. As schematically depicted in Figure 18, the condition of circumferential plasma flow in this direction (arrow 214) continually pulls the interface 58 back to the PRP collection region 76, where the radial plasma flow conditions, upper, described already exist to sweep even more platelets from the 58th interface. Simultaneously, the counterflow patterns serve to circulate the other heavier components of the 58th interface (the lymphocytes, monocytes and granulocytes) back to the mass of the PRBC , far from the PRP stream. Within this circumferential, dynamic plasma flow condition, the MNCs (designated as such in Figure 18) initially settle along the high-G wall 24, but eventually float to the surface of the interface 58 near the region 50 of collection of high hematocrit PRBC. The low taper wall G creates the plasma counterflow patterns, shown by the arrows 214 in Figure 18. These counterflow patterns 214 draw the MNC back to the PRP collection region 76, high. hematocrit. The MNCs are relocated again from the 76 low-hematocrit PRP collection region, to the 24 high-G wall. The MNCs circulate on this route, designated 216 in Figure 18, while the WB is separated into PRBC and PRP. The MNCs are controlled in this manner and "flanked" on this confined route 216 within compartment 38 away from both the PRBC collection region 50 and the PRP collection region 76. Additional details of the separation dynamics are found in processing compartment 38 in U.S. Patent No. 5,573,678, which is incorporated herein by reference.

V. Process of Mononuclear Cell Processing The centrifuge 10 includes a process controller 222 (see Figure 19), which commands the operation of the fluid circuit 200 to perform a pre-set MNC collection and pick-up processing 224 using the container 14.

As shown in Figure 20, processing 224 comprises a pre-processing priming cycle 226, which primes the fluid circuit 200. The processing 224 then includes a preliminary processing cycle 228, which processes the PPP of the whole blood obtained from the donor / patient for later use in the 224 procedure as a means of suspension for the collected MNCs. The processing 224 then includes at least one main processing cycle 230. A main processing cycle 230 comprises a collection step 232, followed by a collection lid 234. Harvesting stage 232 includes a series of harvesting phases 236 and 238, during which the whole blood is processed to accumulate mononuclear cells in the first compartment 38 in the manner previously described. The collection stage also includes a series of collection phases 240, 242, 244 and 246, during which the accumulation of the mononuclear cells is transferred from the first compartment 38 to an MNC collection vessel coupled to the 200 circuit. from suspension collected during the preliminary processing cycle 228, is added to the MNCs. Usually, the main processing cycle 230 will be carried out at one time during a given procedure 224. The number of processing cycles 230 carried out in a given processing 224 will depend on the total volume of MNC that is sought and that is collected. For example, in a representative procedure 224, five main processing cycles 230 are repeated, one after the other. During each cycle 230 of the main processing, from about 1500 to about 3000 ml of whole blood can be processed, to obtain a volume of MNC per cycle of about 3 ml. At the end of the five processing cycles 230, an MNC volume of about 15 ml can be collected, which is dispersed in a final dilution of PPP of about 200 ml.

A. Pre-Proceed Fattening / Ballast Sequence Before a donor / patient is coupled to the fluid circuit 200 (via the TI and T9 tubing), the controller 222 carries out a priming cycle 228. During the priming cycle 228, the controller 222 commands the centrifuge 10 to rotate the spool and cup elements 18 and 20 about the axis 28, while ordering the pumps Pl a P6 transporting a sterile priming liquid, such as saline, from the PRIME (HEAD) and anticoagulant container from the ACD vessel to the entire length of the complete fluid circuit 15, and the container 14. The priming fluid displaces the air of the circuit 15 and the container 14. The second compartment 40 is served by a single pipe T18 and therefore has, in effect, an individual access hole. To achieve the priming, the compartment 40 is isolated from the communication for flow with the priming liquid, while the pump P5 is operated to remove air from the compartment 40, thereby creating a condition of relative pressure (vacuum) in the compartment 40. In the removal of the air from the compartment 40, then communication is opened to the flow of the priming liquid, which is drawn into the compartment 40 by the vacuum. Pump C5 is also operated to assist in the transport of liquid to compartment 40 to create a condition of positive pressure in the compartment 40. A controller 222 retains priming liquids in the second compartment 40, to counterbalance the first compartment 38 during blood processing. Of course, it should be appreciated that this vacuum yeast process is applicable to the priming of virtually any container served by an individual access hole or its equivalent.

Preliminary Processing Cycle The MNCs that are collected in the MNC container are dispersed in a platelet-poor plasma (PPP) medium of the MNC donor / patient.

During the preliminary processing cycle 228 the controller 222 configures fluid circuit 222 to collect a preset volume of PPP from the donor / patient for retention in the container PPP This volume is then used as a suspension medium for the MNCs during processing, as well as being added to the MNCs after processing to achieve the desired final dilution volume.

Once the donor / patient has been phlebographed, the controller 222 configures the pump stations PSL, PSM, and PSR to begin the preliminary processing cycle 2228. During this cycle 228, the whole blood is centrifuged in the compartment 38 in red, packed blood cells (PRBC) and platelet-rich plasma (PRP), as described above. The PRBCs are returned to the donor / patient, while mononuclear cells accumulate in compartment 38. As MNCs accumulate in compartment 38, a portion of the separated plasma component is removed and collected for use as a means of MNC suspension. During cycle 228, controller 222 maintains interface 58 at a relatively low position on ramp 84 (as shown in Figure 16B). as a result, the plasma transported from compartment 38 and stored in the PPP vessel is relatively poor in platelets, and thus can be characterized as PPP. The rest of the PPP transported from compartment 38 is returned to the donor / patient during this cycle 228.

The configuration of the fluid circuit 200 during the preliminary processing cycle 228 is shown in Figure 21, and is further summarized in Table 2.

Table 2 Additional Processing Cycle where: • indicates a closed condition or occluded pipe. or indicates a non-occluded pipe or open condition. * Indicates a pump in condition, during which the pump rotors rotate and couple the Pump tubing to transport fluid in a peristaltic manner. B o Indicates a pump condition turned off, open, during which pump announcers are not rotating and in which the pump rotors do not couple the pump tubing circuit, and therefore allow fluid flow through the pump tubing circuit. B • Indicates an off, closed pump condition, during which the pump rotors are not rotating, and in which the pump rotors are coupled with the pump tubing circuit, and therefore do not allow fluid flow through the pump pipe circuit. During the preliminary cycle 228, the pump P2 draws whole blood (WB) from the donor / patient through the TI pipe in the left cartridge 23L, to the pipe T3, through the chamber DI, and to the blood processing compartment 38 through the pipe T4. The pump P3 draws the anticoagulant ACD through the pipe T15, to the intermediate cartridge 23L and to the pipe T2, for mixing with the whole blood.

Whole blood, anticoagulated, is transported to compartment 38 through hole 48. Whole blood is separated into PRP, PRBC, and the entrecara (including MNC), as previously described. The orifice 50 transports the PRBC 96 through the blood processing compartment 38, through the pipe T5 to the intermediate cartridge 23M. The PRBCs enter the T7 pipe through the T8 pipe, to return the donor / patient via the left cartridge 23L and the T9 pipe. The orifice 46 transports PPP from the blood processing compartment 38. The PPP follows the UNC pipe up to the 23R cartridge. The T5 pump transports a portion of the PPP to the T7 pipe for return with the PRBCs to the donor / patient. The gauge controller 220 adjusts the flow rate of the pump P5 to maintain the gauge at a low position on the ramp 84 (as shown in FIG. 16B) to thereby minimize the concentration of platelets carried during the compartment 38 during this cycle. The P6 pump transports a portion of the PPP through the T12 pipe to the PPP container, until the pre-set volume for MNC suspension and the final solution is collected. This volume is designated VOLsus.

C. Main Processing Cycle 1. Mononuclear Cell Collection Stage (MNC) (i) MNC Accumulation Phase The controller 222 now switches to the pickup stage 232 of the MNC of the main processing cycle 230. First, the controller 222 configures the fluid circuit 200 for the MNC accumulation base 236. For step 236, controller 222 changes the configuration of the PSR pump station to retain the PPP collection. The controller 222 also commands the interleaver controller 220 to maintain a flow rate for the pump P5 to maintain the interlock at a higher location on the ramp 84 (as shown in FIG. 16A), thereby allowing the separation of PRP. Due to the changed configuration, the P6 pump also recirculates a portion of the PRP to the blood processing chamber 38 for improving platelet separation efficiencies, as will be described in detail below. The configuration for the MNC accumulation phase 236 of the MNC collection step 232 is shown in Figure 22, and is further summarized in Table 3.

Table 3: Condition of Mononuclear Cell Collection (MNC Accumulation Phase) where: • indicates a closed condition or occluded pipe. or indicates a non-occluded pipe or open condition. It indicates a pump in condition, during which the pump rotors rotate and couple the pump tubing to transport fluid in a peristaltic manner. ßo Indicates an open, open pump condition, during which pump announcers are not turning and in which the pump rotors do not couple the pump tubing circuit, and therefore allow fluid flow through the pump circuit. pump tubing B »Indicates an off, closed pump condition, during which the pump rotors are not rotating, and in which the pump rotors are coupled with the pump tubing circuit, and therefore do not allow fluid flow through the pump pipe circuit. 1. Promotion of High Efficiencies of Separation of Platelets by the Recirculation of PRP Normally, platelets are not collected during MNC processing. Instead, it is believed that it is desirable to return them to the donor / patient. A high average platelet volume, MPV (expressed in fentoliters, fl, or cubic microns) for separate platelets is desirable, since it denotes a high efficiency of platelet separation. The MPV can be measured by conventional techniques-from a sample of PRP. Larger platelets (greater than about 20 fentoliths) are probably the ones that are caught in the 58th interface and do not enter the PRP to return to the donor / patient. These result in a reduced population of major platelets in the PRP, and therefore a lower MPV, for return to the donor / patient. The establishment of radial plasma flow conditions, sufficient to leave larger platelets of the interface 58, as previously described, is highly dependent on the input hematocrit Hi that enters the blood processing compartment 38. For this reason, the pump 6 recirculates a portion of the PRP flowing in the UNC pipe back to the inlet hole WB. The recirculating PRP flows through the right cartridge 23R to the Til pipe, which joins the pipe T4 coupled to the inlet 48. The recycle PRP is mixed with the WB entering the blood processing compartment 38, thereby decreasing the input hematocrit Hi. The controller adjusts a recirculation flow rate of PRP, QRec? Rc for pump P6 to achieve a desired input hematocrit H ±. In a preferred implementation, the Ex is not greater than about 40%, and more preferably, is about 32%, which will achieve a high MPV. The input hematocrit H ± can be measured conventionally by an in-line sensor in line T4 (not shown). The input hematocrit Hi can also be determined empirically based on the perceived flow conditions, as described in US Patent Application Serial No. 08 / 471,883, which is incorporated herein by reference. 2. Promotion of High Concentration of MNC and Purity by Recirculation of PRBC As shown schematically in Figure 18, the counterflow of the plasma (arrows 214) in the compartment 38 drags the interface 58 back to the PRP collection region 76, where the improved radial plasma flow conditions sweep the platelets of the 58th interface for return to the donor / patient. Counterflow patterns 214 also recirculate other heavier components of the 58th interface, such as lymphocytes, monocytes and granulocytes back for circulation in the PRBC mass. Meanwhile, due to the relatively high hematocrit in the PRBC collection region 80, the MNCs float near the 80 region to the surface of the 58th interface. Here, the MNCs are dragged by the plasma counterflow to the collection region 214. 76 of PRP, low hematocrit. Due to the low hematocrit in this region 76, the MNCs are clearly re-edged towards the high-G wall 54. Arrow 216 in Figure 18 shows the desired recirculation flow of the MNCs as they accumulate in compartment 38. Maintenance of a HBC output hematocrit of PRBC, desired, in region 50 of collection of PRBC is important. If the output hematacrit H0 of the PRBC drops below a low threshold value, given (for example, below about 60%), most MNCs will not circulate as a cell mass, as shown by arrow 216 in Figure 18. Exposed to a low H0, all or some of the MNCs will fall to float toward the 58th interface. Instead, the MNCs will remain banded along the high G wall and transported out of compartment 38 with the PRBC. It results in insufficient MNC performance. On the other hand, if the H0 exceeds a high threshold value, given (eg, about 85%), larger numbers of heavier granulocytes will float on the 58th interface. As a result, the smaller granulocytes will be transported away from the interface 58 for the return with the PRB to the donor / patient. In contrast, most granulocytes will occupy the 58th interface and contaminate the MNC. For this reason, during the MNC collection step 232, the process controller 222 commands the pump P4 to recirculate a portion of the PRBCs flowing in the T5 pipe back to the WB inlet 48. As shown in Figures 21 and 22, the recirculation PRBCs flow through the intermediate cartridge 23M to the pipe T6, which joins the pipe T4 coupled to the inlet 48. The recirculation PRBCs are mixed with the WB entering the blood processing compartment 38. Generally speaking, the magnitude of the output hematocrit H0 varies inversely as a function of the PRBC recirculation flow rate, Qr, which is governed by the pump. P4 (PRBC) and pump P2 (WB). Given a flow rate for the WB established by pump P2, the output hematocrit H0 can be increased by decreasing the Qr, and conversely, the output hematocrit H0 can be decreased by increasing the Qr. The exact relationship between Qr and H0 takes into account the centrifugal acceleration of the fluid in compartment 38 (governed by the magnitude of the centrifugal force in compartment 38), the area of compartment 38, as well as the inflow rate of whole blood (Qb) in compartment 38 (governed by pump P2) and output flow rate PRP (QP) of compartment 38 (governed by the pump P5 of control of entrecara). There are several ways to express this relationship and thus quantify the Qr based on a desired HQ. In the illustrated embodiment, controller 222 periodically samples Qb, Qp, and Qr.

In addition, taking into account active centrifugal force bills in compartment 38, the controller derives a new recirculation pump speed of PRBC Qr (NEW) for pump P4, based on a target H0, as follows: (i) ) Start at the sampling time n = 0 (ii) Calculate the QE as follows: Q = [Q -Q 1 «• [- where: H0 is the output, target hematocrit value, expressed as a decimal (for example, 0.75 for 75%). a is the acceleration of the fluid, governed by the centrifugal forces, calculated as follows: a = r __ * where: O is the rotation speed of compartment 38, expressed in radians per second. r is the radius of rotation. g is unit gravity, equal to 981 cm / sec2. A is the compartment area 38. k is the hematocrit constant and m is a separation performance constant, which is derived based on empirical data and / or theoretical modeling. The preferred modality, the following theoretical model is used: •• Jl - H) to A C. where 1. 08 S and where: ß is a term sensitive to constant effort defined as: ß = 1 + _____ and where: Based on the empirical data, b = 6.0 s_n and n = 0.75, and the constant stress velocity is defined as: du / dy where (u) is the fluid velocity and (y) is a spatial dimension. And where: Sr is a red blood cell sedimentation factor, empirically derived, which, in empirical data, can be established at 95 x 10"9 s This model is based on equation (19) of Brown, "The Physics of Continuous Flow Centrifugal Cell Separation", artificial Organs; 13 (l): 4-20, Raven Press, Ltd., New York (1989) (the "Brown article"), which is incorporated herein by reference. The graph of the model appears in Figure 9 of the Brown Article. The previous model is linearized using a simple linear regression over a range of practical, expected, conditions of blood processing. Algebraic substitutions are made based on the following expressions: * A = B0Q0 where: - Q0 is the PRBC flow velocity through the outlet pipe T5, which can be expressed as: This linearization produces a simplified curve in which the value of (m) constitutes the slope of the value of (k) constitutes the intersection in y-. In the simplified curve, the slope (m) is expressed as follows: Tx = 338.3 (_) S where: ß / Sr can be expressed, based on empirical data, as a constant value of 1.57 / μs.

Therefore, in the simplified curve, 'm has a value of 531.13. A range of values for m between about 500 and about 600 is believed to be applicable to whole blood, centrifugal, continuous blood separation procedures in general. For the simplified curve, the value of the intersection in y for (q) is equal to 0.9489. A range of values for k between about 0.85 and about 1.0 is believed to be applicable to whole blood, centrifugal, continuous blood separation procedures in general. (iii) Calculate the average Qr Qr is measured at selected intervals, and these instantaneous measurements are averaged during the processing period, as follows: Qr (AVG) = [0.95 (C? R (AVG ^)) + [0.05. Qf] (iv) Calculate new Qr as follows Qr (NEW) = Qr (AVG) * F where: F is an optional control factor, which allows the control of Qr (when F = 1), or disables the control of Qr (when F = 0), or allows a rope climb based on system variations ( when F is expressed as a fraction between 0 and 1). F may comprise a constant, or alternatively, may vary as a function of the processing time, for example, starting at a first value at the beginning of a given procedure and changing a second or more values as the process proceeds. (v) Keep Qr within pre-established limits (for example, between 0 ml / min and 20 ml / min.). IF Qr (NEW) > ml / min. THEN Qr (NEW) = 20 ml / min. END IF Qr (NEW) < 0 ml / min. THEN Qr (NEW) = 0 ml / min. END n = n + 1 During the MNC pickup step 232 (Figure 22), the controller 222 simultaneously adjusts and maintains multiple flow rates to achieve optimum processing conditions in the compartment 38 for the accumulation of high high performance MNC purity. The controller establishes and maintains the input flow rate of WB Qb (via pump P2), the output flow rate PRP, Qp (via pump P5), the recirculation flow rate PRP QRec ± rc (via the pump Pß), and the recirculation flow rate PRBC Qr (via pump P4). Given an input flow rate of WB, Qb, which is typically established for the comfort of the donor / patient and the achievement of an acceptable processing type, the controller 222: (i) commands the pump P5 to maintain a set Qp to sustain a desired interface position on the ramp 84, and thereby achieving the desired platelet concentrations in the plasma (PPP or PRP); (ii) order the pump P6 to maintain a QReci c set to retain the desired input hematocrit H ± (eg, between about 32% and 34%), and thereby achieve high platelet separation efficiencies; and (iii) the pump P4 commands to maintain a set Qr to sustain a desired output hematocrit H0 (eg, between about 75% to 85%), thereby preventing granulocyte contamination and maximum MNC yields. (ii). Second Phase (Collection of PRBC) The controller 222 terminates the MNC accumulation phase 236 when a pre-set volume of whole blood is processed (e.g., 1500 mi to 3000 mi). Alternatively, the MNC accumulation phase can be determined when a target volume of MNC is collected. The controller 22 then introduces the collection phase of PRBC 238 of the collection stage of MNC 232. In this phase 238, the configuration of the PSM pump station is altered to retain the return of the PRBCs to the donor / patient (when closing V14), stop the recirculation of the PRBC (when closing valve V18 and place pump P4 in a pump condition shut down, closed, and instead transport the PRBCs to the PRBC vessel (when opening V15). This new configuration is shown in Figure 23, and is further summarized in Table 4.

Table 4: Mononuclear Cell Collection Stage (PRBC Collection Phase) where: • indicates a closed or occluded pipe condition or indicates an unoccluded pipe or open condition.

Indicates a pump-on condition, during which the pump rotors rotate and couple the pump tubing to transport fluid in a peristaltic manner. Bo Indicates an open, open pump condition, during which pump announcers are not rotating and in which the pump rotors do not couple the pump tubing circuit, and therefore allow fluid flow through the pump circuit. pump tubing B "Indicates a pump-off condition, closed, during which the pump rotors are not rotating, and in which the pump rotors are coupled with the pump tubing circuit, and therefore do not allow fluid flow through the pump pipe circuit. In this step 238, the PRBCs on the T45 line are transported through the intermediate cartridge 23M on the line T14, and up to the PRBC container. The controller 222 operates in this phase 238 until a desired volume of PRBC (e.g., 35 mi to 50 ml) is collected in the PRBC vessel. This volume of PRBC is subsequently used in the phase 240 of MNC removal of step 234 of MNC collection, as will be described in detail later. The controller 222 terminates the PRBC pickup phase 238 upon perceiving gravimetrically, using the weight scale WS) that the PRBC container or retains the desired volume of PRBC. The end of step 232 of MNC collection of the main processing cycle 230. 2. Stage of Mononuclear Cell Collection (i) First Phase (Removal of MNC) The controller 222 introduces the MNC collection step 234 of the main processing cycle 230. In the first phase 240 of this step 234, whole blood is withdrawn and recirculated back to the donor / patient without passage through the compartment 38 of blood processing. The PRBCs collected in the PRBC container in the collection phase 238 of the preceding PRBC are returned to processing compartment 38 through the inlet pipe T4 of WB, while the rotation of compartment 38 continues. The MNCs accumulated in compartment 38 during step 232 of MNC collection are transported with the PRP through the UNC pipe out of compartment 38. The fluid circuit configuration 15 during phase 240 of removal of M_STC from step 234 of MNC collection is shown in Figure 24A, and is further summarized in Table 5: Table 5: Mononuclear Cell Collection Stage (MNC Removal Phase) where: • indicates a closed or occluded pipe condition or indicates an unoccluded pipe or open condition. • Indicates a pump on condition, during which the pump rotors rotate and couple the pump tubing to transport fluid in a peristaltic manner. Bo Indicates an open, open pump condition, during which pump announcers are not rotating and in which the pump rotors do not couple the pump tubing circuit, and therefore allow fluid flow through the pump circuit. pump tubing B »Indicates an off, closed pump condition, during which the pump rotors are not rotating, and in which the pump rotors are coupled with the pump tubing circuit, and therefore do not allow fluid flow through the pump pipe circuit. As shown in Figure 24A, the controller 222 closes the PRBC output pipe T5 while the PRBCs are transported by the pump P4 from the PRBC vessel through the pipe T14 and T6 towards the pipe T4, for introduction into the compartment 38 through the inlet 48 of WB. The controller 222 initiates a cycle time counter to TCYCSTAORT. The influx of PRBC from the PRBC vessel through the entry hole WB increases the hematocrit in the PRP collection region 76. In response, the concentrated region of MNC accumulated in compartment 38 (as shown in Figure 18), floats to the surface of the 58th interface. The volume of incoming PRBC moves the PRP through the PRP output orifice 46. The interface 58, and with it, the concentrated MNC region (designated MNC region in Figure 24A) also travels out of the compartment 38 through the PRP exit orifice 46. The MNC region moves along the PRP UNC pipe to the optical OS sensor. As shown in Figure 28, within the UNC pipe, a PRP region 112 precedes the concentrated MNC region. The PRP in this region 112 is transported to the PPP vessel through the right cartridge 23R of pipe T12 (as shown in Figure 24A). A region 114 of PRBC it also follows the MNC region concentrated within the TÍO pipeline. A first transition region 116 exists between the PRP region 112 and the MNC region concentrated. The first transition region 116 consists of a regularly decreasing concentration of platelets (shown by a square pattern in Figure 28) and a regularly increasing number of MNCs (shown by a textured pattern in Figure 28). A second transition region 118 exists between the concentrated MNC region and the PRBC region 114. The second transition region 118 consists of a regularly decreasing concentration of MNC (shown by a textured pattern in Figure 28) and a regularly increasing number of PRBC (shown by a wavy pattern in Figure 28). Viewed by the optical sensor, the regions 112 and 116 that precede the MNC region and the regions 118 and 114 that follow the MNC region exhibit a transition optical density at which the MNC region can be discerned. The optical sensor OS perceives the changes in optical density in the liquid transported by the UNC pipe between the exit port 46 of PRP of the right cartridge 23R. As shown in Figure 28, the optical density will change from a low value, indicating that it is highly transmitting to light (i.e., in the PRP region 112), at a high value, indicating that it is highly absorbent to light (it is say, in PRBC region 114), as the MNC region proceeds beyond the optical OS sensor. In the embodiment illustrated in Figure 28, the optical OS sensor is a conventional hemoglobin detector, used, for example, in the Autopheresis-CR blood processing device sold by the Fenwal Division of Baxter Healthcare Corporation. The OS sensor comprises a red light emitting diode 102, which emits light through the UNC pipe. Of course, other wavelengths, similar to green or infrared, could be used. The OS sensor also includes a PIN diode detector 106 on the opposite side of the UNCLE pipe. The controller 222 includes a processing element 100, which analyzes the voltage signals received from the emitter 102 and the detector 106 to compute the optical transmission of liquid in the UNCLE pipeline, which is called OPTTRANS.

Several algorithms can be used by the processing element 100 to compute the OPTTRANS. For example, the OPTTRANS can match the output of the diode detector 106 when the red light emitting diode 106 is on and the liquid flows through the UNCLE (RED) pipe. The background optical "fluid" can be filtered from the network to obtain OPTTRANS, as follows: OPTTRANS = 8R l RED SPILL) CORRREF where COR (RED SPILL) is calculated as follows COR (RED SPILL) = RED-REDBKGRD where: RED is the output of the diode detector 106 when the red light emitting diode 102 is on and the filter flows through the UNC pipe.

REDBKGRD is the output of the diode detector 106 when the red light emitting diode 102 is turned off and the liquid flows through the UNC pipe; and where CORREF is calculated as follows: CORREF = REF-REFBKGRD where: REF is the output of the red light emitting diode 102 when the diode is on; and REFBKGRD is the output of light emitting diode 102 when the diode is off. The processing element 100 normalizes the OS sensor to the optical density of the PRP of the donor / patient, by obtaining the OS sensor data during the preceding MNC collection step 232, as the PRP of the donor / patient is transported through the donor. the UNCLE pipe. These data establish a baseline optical transmission value for the pipeline and the PRP of the donor / patient (OPTTRANSBASE) • For example, the OPTTRANSBASE can be measured at a selected time during the collection stage 232, for example, halfway through the stage 232, using either a scheme of filtered or unfiltered detection, as described above. Alternatively, a set of optical transmission values is calculated during step 232 of MNC collection using either a filtered or unfiltered detection scheme. The set of values is averaged during the complete collection stage to derive OPTTRANSBASE • The processing element 100 continues during the MNC removal phase 240, subsequent to perceiving one or more optical transmission values for the UNC pipeline and the flowing liquid in this one (OPTTRANSHARVEST) during phase 240 of MNC removal. The OPTTRANSHARVEST may be comprised of an individual reading sensed at a selected time of phase 240 of MNC removal (eg, the law of way through phase 240), or may comprise an average of multiple readings formed during phase 240 of removal of MNC. The processing element 100 derives a normalized DENSITY value by setting OPTTRANSBASE to 0.0, setting the optical saturation value to 1.0, and adjusting the value of OPTTRANSHARVEST proportionally in the normalized value range of 0.0 to 1.0.

As shown in Figure 28, the processing element 100 retains two predetermined threshold values THRESH (l) and THRESH (2). The value of THRESH (l) corresponds to a nominal value selected for DENSITY (for example, 0.45 on a normalized scale of 0.0 to 1.0), which has been empirically determined to occur when the concentration of MNC in the first transition region 116 satisfies a pre-selected processing target. The value of THRESH (2) corresponds to another nominal value, selected for DENSITY (for example, 0.85 on a normalized scale from 0.0 to 1.0), which has been empirically determined to occur when the concentration of the PRBCs in the second transition region 118 exceeds the pre-selected processing target. The liquid volume of the UNC pipe between the optical sensor OS and the valve station V24 in the right cartridge 23R constitutes a known value, which is introduced to the controller 222 as a first volume of misalignment VOLOFFC? • The controller 222 calculates a first control time value Time! based on VOLOFF.U and pump speed of pump P4 (Qp) as follows: VOL Time, rrxx i * 60 eP- In the illustrated and preferred embodiment, the operator can specify and input to the controller 222 a second volume of misalignment VOLOFF.2./- which represents an additional, nominal volume (shown in Figure 28) to increase the collected volume of MNC, total , VOLMNC- The amount of VOL0FF (2) takes into account the variations of the processing system, as well as the variations between the donors / patients in the purity of the MNCs, The controller 222 calculates a second value of control time TIME2 in base a VOL0FF (2) and the pump speed of pump P4 (QP4)) as follows: Time, = VOL ° r p x? 60 As the operation of the pump P4 transports the PRBC through the inlet port 48 WB, the interface 58 and the MNC region advance through the PRP UNC pipe to the sensor optical OS. The PRP that precedes the MNC region advances beyond the optical OS sensor, through the T12 pipe, and to the PPP vessel. When the MNC region reaches the optical OS sensor, the OS sensor will sense the DENSITY THRESH (l). In this case, the controller 222 initiates a first timer TCi. When the optical sensor senses DENSITY = THRESH (2) the controller 222 initiates a second timer TC2. The perceived MNC volume can be derived based on the interval between TCi and TC2 for the given QP4. As time progresses, the controller 222 compares the magnitudes of TCX to the first control time Ti, as well as compares the TC2s to the second control time T2. When TCX = TCi, the leading edge of the target MNC region has arrived at the valve station V24, as shown in Figure 24B. The controller 222 commands the valve station V24 to open, and commands the valve station V25 to close, the controller 222 marks this event in the cycle time counter as TCYCS? TcH "The target MNC region is transported in the pipe T13 leading to the container MNC. When TC2 = T2, the second misalignment value V0LOFF.2. It has also been transported in pipe T13, as shown in Figure 24C. The total volume of MNC selected for collection (V0LMNC) for the given cycle is thus present in pipe T13. When TC2 = T2, the controller 222 commands the P4 pump to stop. The further advancement of V0LMNC in theory T13 therefore ceases. The controller 222 derives the volume of PRP that was transported to the PPP vessel during the preceding MNC removal phase. This volume of PRP (which is designated V0LPRP) is derived, as follows: CYC - CYC - ^ PRP °.

In a preferred embodiment, the controller 222 terminates the MNC removal phase, independent of TCi and TC2 when pump P4 transports more than one volume of specified PRBC fluid after TCYCSTART (eg, more than 60 ml). This time-out circumstance may occur, for example, if the optical OS sensor fails to detect the THRESH (l). In this circumstance of time out, volumetric, VOLPRP = 60 - VOLOFFCL ..

Alternatively, or in combination with a tie, mpo outside volumetric, the controlled 222 can determine the MNC removal phase independent of TCi and TC2 when the WS weight scale for the PRBC vessel perceives a weight less than a pre-established value ( for example, less than 4 grams, or the equivalent in weight of a volume of fluid less than 4 ml). (ii) Second Phase (Flood of PRP; Once the MNC region is placed as shown in Figure 24C, the controller 222 introduces the PRP flood phase 242 of the MNC collection step 234. During this phase 242, the controller 222 configures the circuit 200 to move the VOLPRP out of the PPP container and the T12 pipe and into the blood processing compartment 38. The configuration of the fluid circuit 200 during the flood phase 242 of PRP is shown in Figure 25, further summarized in Table 6.

Table 6: Mononuclear Cell Collection Stage (PRP Elimination Phase) where: • indicates a closed condition or occluded pipe. or indicates a non-occluded pipe or open condition. • Indicates a pump on condition, during which the pump rotors rotate and couple the pump tubing to transport fluid in a peristaltic manner.

Bo Indicates an open, open pump condition, during which pump announcers are not rotating and in which the pump rotors do not couple the pump tubing circuit, and therefore allow fluid flow through the pump circuit. pump tubing B »Indicates an off, closed pump condition, during which the pump rotors are not rotating, and in which the pump rotors are coupled with the pump tubing circuit, and therefore do not allow the flow of fluid through the pump tubing circuit. During the PRP flood stage 242, the controller 222 configures the pump stations PSL, PSM and PSR to stop the recirculation of whole blood, and while the rotation of the compartment 38 continues, to pump VOLPRP to the processing compartment 38 to through the Til pipe. The VOLPRP is transported by the pump P6 through the pipe T12 to the right cartridge 23R, and from there to the pipe Til, for entry into the processing compartment 38 through the pipe T4 and the hole 48. The PRBC are transported from the processing compartment 38 through the hole 50 of the T5 pipe to the intermediate cartridge 23M, and from there to the pipes T8 and T7 in the left cartridge 33L. The PRBCs are transported to the T9 pipeline for return to the donor / patient. No other fluid is transported in the fluid circuit 15 during this step 42. The return of VOLPRP restores the volume of the liquid in the PPP to VOLsus vessel, as it is collected during the preliminary processing cycle 228 described previously. The return of VOLpRp also retains a low population of platelets in the VOLsus in the PPP vessel recorded for MNC suspension. The return of VOLpRp also transports the residual MNC present in the first transition region 116 before TCi = Ti (and therefore not part of VOLMNC) .- back to processing compartment 38 for additional collection in a 230 cycle of main processing, subsequent. (iii) Third Phase (Suspension of MNC) With the return of V0LPRP to the compartment 38, the controller 222 introduces the MNC suspension phase 244 of the MNC collection step 234. During this phase 244, a portion of V0LSUs in the PPP vessel is transported with VOLMNC in the MNC vessel. The configuration of the fluid circuit 200 during the suspension phase 244 of MNC is shown at 26, and is further summarized in Table 7.

Table 7: Mononuclear Cell Collection Stage (MNC Suspension Phase) where: • indicates a closed or occluded pipeline condition or indicates a non-occluded pipe or open condition. Indicates a pump-on condition, during which the pump rotors rotate and couple the pump tubing to transport fluid in a peristaltic manner. Bo Indicates an open, open pump condition, during which pump announcers are not rotating and in which the pump rotors do not couple the pump tubing circuit, and therefore allow fluid flow through the pump circuit. pump tubing B "Indicates a pump-off condition, closed, during which the pump rotors are not rotating, and in which the pump rotors are coupled with the pump tubing circuit, and therefore do not allow fluid flow through the pump pipe circuit. In phase 244 of MNC suspension, the controller closes C3 to stop the return to PRBC to the donor / patient. A predetermined aliquot of V0LSUs (for example, from 5 ml to 10 ml) is transported by pump P6 through line T12 in right cartridge 23R and then to line T13. As shown in Figure 26, the aliquot of VOLsus further advances VOLMNC through the pipe T13 IN the MNC vessel. (iii) Fourth Phase (Cleaning) At this time, the controller 222 introduces the final cleaning phase 42 of the MNC collection step 234. During this phase 246, the controller 222 returns the PRBCs resident in the UNC pipe to the processing compartment 38. The fluid circuit configuration 200 during the cleaning phase 246 is shown in Figure 27, and is further summarized in Table 7. .

Table 7: Mononuclear Cell Collection Stage (Cleaning Phase) where: • indicates a closed condition or occluded pipe. or indicates a non-occluded pipe or open condition. Indicates a pump-on condition, during which the pump rotors rotate and couple the pump tubing to transport fluid in a peristaltic manner.

Bo Indicates an open, open pump condition, during which pump announcers are not rotating and in which the pump rotors do not couple the pump tubing circuit, and therefore allow fluid flow through the pump circuit. pump tubing B »Indicates an off, closed pump condition, during which the pump rotors are not rotating, and in which the pump rotors are coupled with the pump tubing circuit, and therefore do not allow fluid flow through the pump pipe circuit. The cleaning phase 246 returns any residual MNC present in the second transition region 118 (see Figure 28) after TC2 = T2 (and therefore not part of VOLSEN), back to the processing compartment 38 for additional collection in a subsequent processing cycle. In the cleaning phase 246, the controller 222 closes all the valve stations in the intermediate left cartridges 23L and 23M and configures the right pump station PSR to circulate the PRBCs from the UNC pipe back to the processing compartment 38 through the Tubing Til and T4. During this period, components are not being withdrawn, or returned to the donor / patient. At the end of the cleaning phase 246, the controller 222 starts a new main processing cycle 230. The controller 222 repeats a series of processing cycles 230 until the desired volume of the target MNC for the complete process is reached. At the end of the last main processing cycle 230, the operator may desire the additional VOLsus to further dilute the MNCs collected during the procedure. In these circumstances, the controller 222 may be arranged to configure the fluid circuit 200 to perform a preliminary processing cycle 228, as described above, to collect additional VOLSUs from the PPP vessel. The controller 222 then configures the fluid circuit 200 to carry out a suspension phase of MNC 244, to transport additional VOLsus in the MNC vessel to achieve the desired dilution of VOLMNC.

IV. Alternative Processing of Mononuclear Cells Figure 29 shows an alternative embodiment of a fluid circuit 300, which is suitable for collecting and collecting the MNCs. Circuit 300 in many aspects is the same as circuit 200, shown in Figure 6, and common components are given with the same reference numbers. The circuit 300 differs from the circuit 200 in that the second compartment 310 of the container 14 is identical to the compartment 38, and therefore comprises by itself a second blood processing compartment with the same characteristics as the compartment 38. The compartment 310 includes stamps interiors, as shown for compartment 38 in Figure 4, creating the same blood collection regions for PRP and PRBC, the details of which are not shown in Figure 29. Compartment 310 includes an orifice 304 for transporting whole blood in the compartment 310, a hole 310 for transporting PRP from the compartment 310, and a hole 302 for transporting the PRBCs from the compartment 310. Compartment 310 also includes a used ramp 84, as shown in Figures 16A and 16B and as described above in conjunction with compartment 38. Fluid circuit 300 also differs from fluid circuit 200 in that the pipes T14, T18, T19 are not included. In addition, the PRBC container is not included. In contrast, the fluid circuit 300 includes several new pipe routes and clamps, as follows:. Pipe route T21 leads from the PRP outlet hole 306 of compartment 310 through the new clamp C5 to join the pipeline UNCLE. The pipe route T22 leads from the WB inlet hole 306 of the compartment 310 through a new air detector D3 and a new clamp C6 to join the pipe route T3. The pipe route T33 leads from the outlet port PRBC 302 of the compartment 310 through a new clamp C8 to join the pipe T4. The new clamp C7 also provides the T3 pipe upstream of the DI air detector.

The new clamp C9 is also provided in the UNC pipe between the OS optical sensor and the junction of the new T21 pipe. Using the circuit 300, the controller 222 proceeds through the priming cycle 226, previously described, the preliminary processing cycle 228, and the main processing cycle 230 as previously described for the circuit 200, through the phase 236 of MNC accumulation. The PRBC collection phase 238 differs when the circuit 300 is used, since the PRBCs used for the subsequent removal of the MNCs in the compartment 38 are processed and collected in the second compartment 310. More particularly, as shown in FIG. Figure 30, during the PRBC collection phase 238, the controller 222 transports a whole blood volume from the donor / patient to the second compartment 310. The whole blood volume is carried by the P2 pump through the TI pipeline to the pipe T3 and from there through the open clamp C6 to the pipe T22, which leads to the compartment 310. The clamp C7 is closed, to block the transport of the whole blood to its compartment 38, where the MNCs have been accumulated for the collection. The clamp C9 is also closed to block the transport of the PRP from the compartment 38, thereby maintaining the accumulation of the MNCs without disturbance in the compartment 38. In the compartment 310, the whole blood volume is separated into PRBC and PRP, in the same way that these components are separated in the compartment 38. The PRP is transported from the compartment 310 through the pipe T23 and the open clamp C5 by the operation of the pump P5, for the return to the donor / patient. The clamp C8 is closed, to retain the PRBCs in the compartment 310. The controller 222 also performs a different phase 240 of MNC removal, using the circuit 300. As shown in Figure 31, during the phase of removal of MNC 240, controller 222 recirculates a portion of the whole blood withdrawn back to the donor / patient, while directing another portion of the whole blood to compartment 310, following the same route as previously described in conjunction with Figure 30. The controller 222 to the clamps C8 and C9, while closing the clamp C5. The blood The whole entering the compartment 310 displaces the PRBCs through the PRBC outlet hole 302 in up to the T23 pipe. The PRBCs from the compartment 310 enter the entry hole WB of the compartment 38. As described above, the incoming PRBC flow from outside of the compartment 38 increases the PRBC hematocrit within the compartment 38, causing the accumulated MNCs to float to the 58. As described above, the inlet PRBCs from the outside of compartment 38 displace the PRP through the orifice 46 of the PRP, together with the MNC region, shown in Figure 31. In the MNC region it is detected by the optical sensor OS and is collected in the subsequent processing 242, 244 and 246 in the same manner as described for circuit 200. Several features of the invention are set forth in the following claims.

It is noted that in relation to this date, the best method known by the applicant to carry out the present invention, is the conventional one for the manufacture of the objects to which it refers.

Having described the invention as above, the content of the following is claimed as property:

Claims (30)

1. A blood separation system, characterized in that it comprises: a chamber for rotation about an axis of rotation, the chamber including an entrance region where whole blood is introduced for separation into red, packed blood cells having a value of hematocrit HPRBc, a constituent of the plasma, and an interface that has mononuclear cells between the red, packed blood cells and the plasma constituent, and a controller operable in a first mode to transport whole blood to the input region while stirring, the red, packed blood cells and the plasma constituent of the chamber and while maintaining the interface within the chamber, the controller which is also operable in the first mode to maintain an established HPRBC by transporting the red blood cells, packed to the region of entry.

2. A system according to claim 1, characterized in that the controller circulates the blood cells red, packed removed from the chamber for transport to the entrance region during the first mode.

3. A system according to claim 1, characterized in that the whole blood transported to the inlet region has a HWB hematocrit, and wherein the controller is operable in the first mode to maintain an established HWB, by circulating the plasma constituent in the whole blood.

4. A system according to claim 1, characterized in that the controller includes a perception element for locating the interface in the chamber and providing a perceived output.

5. A system according to claim 1, characterized in that the controller is operable, during the first mode, to maintain the enter at an established location, in the base chamber, at least in part, at the perceived exit.

6. A system according to claim 4, characterized in that the perception element optically locates the interface in the chamber.

7. A system according to claim 1, characterized in that the controller is operable in a second mode to remove the chamber interface by removing the red blood cells, packed up to the entrance region.

8. A system according to claim 7, characterized in that, during the second mode, the controller terminates the transport of the whole blood to the entrance region.

A system according to claim 7, characterized in that, during the second mode, the controller terminates the removal of red, packed blood cells from the chamber.

10. A system according to claim 7, characterized in that the controller circulates red, packed blood cells removed from the chamber during the first mode for transport to the entry region during the second mode.

11. A system according to claim 7, characterized in that it also includes a reservoir, and where the controller circulates the red blood cells, packed from the reservoir for transport to the entrance region during the second mode.

12. A system according to claim 11, characterized in that the controller transports blood cells to the reservoir red, packed, removed from the camera during the first mode.

13. A system according to claim 11, characterized in that the reservoir comprises a second chamber rotatable about an axis in which red, packed blood cells are centrifugally separated from whole blood.

14. A system according to claim 7, characterized in that it also includes an exit route for transporting the removed mask of the chamber, which includes a perception element for locating the mononuclear cells, in the removed mask and providing a perceived output in the location of mononuclear cells.

15. A system according to claim 14, characterized in that the perception element optically locates the mononuclear cells in the removed shell.

16. A method for collecting mononuclear cells from whole blood, comprising the steps of: (i) rotating a chamber around a rotational axis, (ii) transporting whole blood in an inlet region of the chamber for separation into blood cells red, packed that have a hematocrit HPRBC value, and a plasma constituent, and an interface that has mononuclear cells between the red, packed blood cells and the plasma constituent, and (iii) during step (ii), remove the red, packed blood cells and the plasma constituent of the chamber, while maintaining the interface within the chamber, and (iv) during step (ii), maintaining an established HPRBc, by transporting the red blood cells, packed up to the region of entry.

17. A method according to claim 16, wherein in step (iv), red blood cells, packed, removed from the chamber in step (iii) they are recirculated to the inlet region.

18. The method according to claim 16, wherein during step (ii), the whole blood that is transported to the input region has a HWB hematocrit, and that further includes, during step (iv), maintaining an established HWB , by circulating the plasma constituent in the whole blood.

19. The method according to claim 16, characterized in that during step (iii), the location of the interface in the chamber is perceived.

20. The method according to claim 16, characterized in that during step (iii), the interface is maintained at a location established in the chamber, based, at least in part, on the perception of the interface.

21. A method according to claim 16, characterized in that during step (iii), the location of the interface in the chamber is optically perceived.

22. A method according to claim 16, and characterized in that it further includes the step of (v) removing the chamber shell by transporting the red blood cells, packed to the entrance region.

23. A method according to claim 22, characterized in that during step (v), the controller terminates the transport of whole blood to the input region.

24. A method according to claim 22, characterized in that during step (v) -, the controller completes the removal of the red blood cells, packed from the chamber.

25. A method according to claim 22, characterized in that during step (v), red, packed blood cells removed from the chamber during step (iii) are transported to the entry region.

26. A method according to claim 22, characterized in that during step (v), the red, packed blood cells are circulated from a deposit for transport to the entry region.

27. A method according to claim 22, characterized in that during step (iii), red, packed blood cells removed from the chamber are transported to the reservoir.

28. A method according to claim 22, characterized in that during step (v), the tank is rotated while the whole blood is transported to the tank to obtain by centrifugal separation the red, packed blood cells, which are transported to the entrance region.

29. A method according to claim 22, characterized in that during step (v), the location of the mononuclear cells in the removed shell is perceived.

30. A method according to claim 22, characterized in that during step (v) the location of the mononuclear cells in the removed shell is optically perceived.