Patent Grant
11660603
In the field of regenerative medicine, access to a broad cross section of sub-dermal tissue is typically required to not only source cells but to also deliver therapy. Fluid tissue that is aspirated or otherwise sourced is often separated into one or more components that are present in the fluid tissue, e.g., plasma, red blood cells, fat cells, stem cells or other nucleated cells. Typically, one or more selected components of the fluid tissue are concentrated into a small volume so that the selected components can be used clinically. For example, there are several commercial devices to separate and concentrate nucleated cells from aspirated bone marrow, fat, or cord blood. Some of these systems employ a floating insert or buoy that is meant to create an interface between the separated fluid components or fractions of interest. The challenge for any apparatus designed to accomplish such a task is the ability to volume reduce the fluid in which the nucleated cells are suspended while recovering as many cells as possible. For example, in marrow aspirate, approximately 1 to 2 percent of the cells suspended in the fluid are the target nucleated cells. Many commercial devices are not able to consistently capture high percentages of nucleated cells while at the same time efficiently volume reduce (i.e., concentrate) the beginning fluid. In other words, many devices are not able to simultaneously obtain a high yield and a high final concentration.
Apparatus and methods for separating components of different densities from a physiological fluid containing cells are described in a previously filed application, International Application No. PCT/US2010/036696, filed on May 28, 2010, published on Dec. 2, 2010 as WO 2010/138895 A2, and incorporated herein by reference in its entirety.
As shown in
To retrieve the separated layers or fluid components, the user takes the syringe or container 2102 out of the centrifuge. As shown in
As shown in
A system for separating components of different densities from a physiological fluid containing cells using a centrifuge includes a container having a top, a sidewall extending from the top, and a bottom disposed opposite the top and in sealing engagement with the sidewall. The container defines a cavity for receiving the fluid. The system includes an insert slidably disposed in the cavity of the container. The insert defines a lumen through the insert, the lumen including a hole and a funnel-shaped upper portion in fluid communication with the hole. The lumen forms an open fluid path between opposite ends of the insert. The insert has a density such that upon centrifugation a selected component of the fluid resides within the lumen. A container port is disposed in the top of the container to transfer the fluid into the container and to withdraw a fluid component other than the selected component from the container. The system further includes a manifold that includes a manifold port, a vent to vent the container, and a connector to couple to the container port. A cannula is receivable in the manifold port and extendable through the container port into the container and into the lumen of the insert to withdraw the selected component from the lumen.
The cannula can include a closed end to close the hole in the insert and a side port to withdraw the selected component. The cannula and the insert may form a seal when the closed end of the cannula closes off the hole in the insert. In an embodiment, the cannula a first cannula, and the system further includes a second cannula extendable through the container port to withdraw the component other than the selected component. The second cannula may be receivable in the manifold port. The system may include two manifolds, each including a manifold port, a vent to vent the container, and a connector to couple to the container port, and the first cannula can be receivable in the manifold port of one manifold while the second cannula can be receivable in the manifold port of the other manifold.
The container can be a syringe and the bottom can be movable, i.e., a plunger, which can have a removable handle. In an embodiment, the plunger is a first plunger and the system further includes a second plunger disposed in the syringe below the first plunger to move the first plunger, for example, to transfer the fluid into the container. The system may further include a clamping mechanism to hold the insert in place after centrifugation, the clamping mechanism being configured to press the sidewall of the container inward against the insert.
A method of separating components of different densities from a fluid containing cells using a centrifuge includes receiving the fluid in a separation system such as the separation system described above, and applying centrifugal force to the separation system. The method further includes, after centrifugation, withdrawing a fluid component other than the selected component through the container port; coupling a manifold to the container port, the manifold including a manifold port and a vent to vent the container; extending a cannula through the container port into the container and into the lumen of the insert, the cannula receivable in the manifold port; and withdrawing the selected component with the cannula from the lumen of the insert.
Withdrawing the selected component may include withdrawing the selected component through a side port in the cannula. In an embodiment, the cannula is a first cannula and withdrawing the component other than the selected component includes extending a second cannula through the container port, the second cannula receivable in the manifold port, and withdrawing the component other than the selected component with the second cannula. The manifold can be coupled to the container before the withdrawing of the component other than selected component. The method may further include with a clamping mechanism, holding the insert in place after centrifugation.
A system for separating components of different densities from a physiological fluid containing cells using a centrifuge includes a container, having a bottom, a top disposed opposite the bottom, and a sidewall extending from the top, the container defining a cavity for receiving the fluid. An insert is slidably disposed in the cavity and defines a lumen through the insert, the lumen including a hole and a funnel-shaped upper portion in fluid communication with the hole. The insert has a density such that upon centrifugation a selected component of the fluid resides within the lumen. The lumen forms an open fluid path between opposite ends of the insert. A container port is disposed in the top of the container. An extraction cap is provided to couple to the top of the container, the extraction cap including a cannula assembly receivable in the container port. The cannula assembly is extendable into the cavity of the container to butt against the insert and to withdraw the selected component from the lumen of the insert.
The cannula assembly can include an inner cannula coaxially disposed within an outer cannula. The inner cannula may include a closed end to close the hole in the insert and a side port to withdraw the selected component, the inner cannula and the insert forming a seal when the closed end of the inner cannula closes off the hole in the insert. The outer cannula may include an open end displaced from the distal end of the cannula assembly to withdraw fluid at a predetermined height above the distal end of the cannula assembly.
In an embodiment, the extraction cap includes a first port in fluid communication with the inner cannula and a second port in fluid communication with the outer cannula. The system may further include a first syringe to couple to the first port and a second syringe to couple to the second port. The cap may include an assembly tab adjacent the first and second ports, the assembly tab extending from the cap to prevent the second syringe from coupling to the first port. The system may further include a lock-out element on the second syringe. For example, the lock-out element includes a tab that locks a plunger of the first syringe until second syringe is removed from the cap. In an embodiment, the extraction cap includes an outer part and an inner part, the inner part carrying the needle assembly and being movable relative to the outer part. The cap may include a locking screw coupled to the outer part and positioned at an angle relative to inner part to push the inner part toward the container with rotation of the locking screw.
A method of separating components of different densities from a fluid containing cells using a centrifuge includes receiving the fluid in a separation system, the system including a container having a bottom, a top disposed opposite the bottom, and a sidewall extending from the top, the container defining a cavity for receiving the fluid. A container port is disposed in the top of the container. An insert is slidably disposed in the cavity of the container, the insert including a funnel-shaped upper portion and a hole therethrough, the insert having a density such that upon centrifugation a selected component of the fluid resides within the upper portion of the insert. The method includes applying centrifugal force to the system and inserting a cannula into the container through the container port to butt against the insert, the cannula having one or more side ports displaced from a distal end of the cannula. The method further includes withdrawing the selected component through the side ports in the cannula; ejecting at least a portion of the withdrawn component through the side ports causing one or more fluid jets in the funnel-shape upper portion of the insert to release cells that adhere to the insert; and withdrawing the ejected portion and cells released by the fluid jets through the side ports.
In any of the systems or methods described herein the insert can be rigid. The volume contained in the lumen upper portion of the insert can be between 5% and 20% of the volume of the container cavity. The selected component (also referred to herein as a target fraction) can be buffy coat and the component other than the selected component can be blood plasma.
Embodiments of the current invention overcome the limitations of known devices for concentration of cells sourced from marrow or other tissue. For example, the insert of the separation device does not form a closed recess or a depression or indent to capture cells, but rather allows for the natural sedimentation of the fluid within the container and through the insert. The insert defines a lumen that has at least one relatively large through hole or channel, including a funnel-shaped upper portion, that allows for the free flow of fluid within the container and through the insert and does not interfere with the natural layering of different density components of the fluid. In addition, the insert identifies the location of a layer of interest, including the target cells. The funnel-shaped upper portion and the through hole reduce the cross-sectional area and increase the thickness of the layer of interest. This facilitates extraction of the target cells and contributes to a high yield and high concentration of the target cells.
Embodiments of the current invention overcome limitations of other systems that use inserts or buoys without a through hole and where the fluid path under centrifugation is confined to the distance between the inner wall of a container or tube and the outer walls of the inserts or buoys. In those systems, minor clots, particles, or other inconsistencies in the fluid can lodge between the walls of the tube and the buoys interfering with the natural layering of the different density components of the fluid. The insert(s) described includes a density selected such that after centrifugation the target cells reside within the hole, the funnel-shape upper portion, or lumen defined by the insert. Under gravitational force the insert floats freely within the container with substantially all of the fluid flowing through the hole or lumen of the insert, but not between the outer wall of the insert and the inner wall of the container. The distance between the inner wall of the container and the outer wall of the insert creates enough space to allow the insert to move freely within the container.
Embodiments of apparatus and methods for separating components of a fluid can be combined with devices and methods to access and source, e.g., aspirate, tissue, such as the aspiration needle assemblies described in International Application No. PCT/US2010/036696. Once tissue is sourced, e.g., loaded into a separation system, the system can be centrifuged. Upon centrifugation, the target cells naturally sediment into the through hole or lumen of the floating insert. These cells are then isolated by means of a cannula. The closed end of the cannula can close the hole in the insert. The target cells residing in the hole of lumen of the floating insert may be sealed from fluid below while fluid above the insert is removed through a cannula. The combination of the cell concentration and separation apparatus described herein with an aspiration apparatus allows a clinician the ability to access subcutaneous tissue in a less traumatic manner and then concentrate nucleated cells from that tissue aspirate. The apparatus can be combined, e.g., coupled or connected, by means of tubing and fluid ports, including luer connections, to create a total solution from aspiration to concentration.
The foregoing will be apparent from the following more particular description of example embodiments of the invention, as illustrated in the accompanying drawings in which like reference characters refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating embodiments of the present invention.
A description of example embodiments of the invention follows.
A prior version of a system for concentrating and separating cells (also referred to herein as a cell concentrator) has been described in International Application No. PCT/US2010/036696, published on Dec. 2, 2010 as WO2010/138895, incorporated herein by reference in its entirety. Here, an improved system is described that has been fabricated and tested. Some key features of the improved system include:
a) a locking base;
b) a double needle extraction system with jet flushing;
c) an extraction needle lock that forces needle to seat in float; and
d) a syringe lock-out tab to ensure proper order of extraction.
To separate buffy coat from red blood cells and plasma, the float 16 should have a density that matches that of the buffy coat, whose density is roughly 1.06 g/cc. A polystyrene material was selected to manufacture the insert because the material's density is close to that of buffy coat. However, testing indicated that a polystyrene float was lighter than desired. Small slugs 56 (
The materials of the system are polycarbonate for most of the parts with the vial being PET (Polyethylene teraphthalate plastic) and the float polystyrene, as described above. Other suitable materials for the float may be polyethylene or polypropylene materials which tend to be less sticky to cells than polystyrenes. However, the densities of these materials are lower than that of buffy coat, so a larger metal material may need to be incorporated into the float to achieve the desired density. Another option is to coat the surface of the float and the inside of the vial with a substance that prevents cells form sticking, for example, a coating from the company Hydromer.
Post centrifugation, the serum (plasma) is above the float 16, the red blood cells are below the float and the target cells (e.g., buffy coat) are inside the float funnel. This separation of fluid components is analogous to what is shown in
As illustrated in
As shown in
To remove the target cells, the extraction cap 18 is placed over the vial 14, as illustrated in
As shown in
Embodiments shown in
A system for separating components of different densities from a fluid containing cells and for concentrating cells according to another example embodiment of the invention is described below and illustrated in
As illustrated in
As illustrated in
A step by step overview of the operation of the system will be described. The first step is to use the double plunger syringe 114 (alternatively, a syringe with a removable handle) to fill the syringe with the blood or marrow specimen to be concentrated.
The second step is to connect to the syringe 114 containing the specimen (e.g., fluid tissue), after the second plunger has been removed, a small micron vented luer cap 178 and then place the syringe inside the syringe tube holder 174. The holder 174 has a solid bottom and also has an O-ring 176 attached to the solid bottom (
Inside of the syringe 114 is a funnel shaped insert 116 (also referred to herein as a funnel) with a hole in the center. The density of the funnel is such that after density separation, target cells from blood or marrow will reside inside of the funnel. Consequently, after density separation of blood or marrow, plasma will reside at the top of the syringe 114 nearest the luer tip (container port) 130, the target cells will reside inside of the funnel 116, and red blood cells will reside beneath the funnel nearest the plunger 113. Two example funnels 116a, 116b are shown in
With respect to the materials used to make the funnel 116 and the shape of the funnel, it should be noted that various materials and shapes can work. When selecting a material, one consideration is whether the funnel 116 is to be molded or machined. In
The third step is to take the syringe 114 and tube holder 174 from the centrifuge, place the syringe inside the clamping mechanism 112 and engage the clamp as illustrated in
The fourth step is to remove the vented cap 178 from the collection syringe 114 and connect to the collection syringe, via the upper luer connection 130, the 30 ml plasma extraction syringe 120 connected to cannula 170 via manifold 160, as shown in
After connection the plasma extraction syringe 120 and cannula 170 to the collection syringe 114, the user pushes the extraction cannula 170 into the collection syringe. The cannula will advance until it hits the funnel 116 that has been frozen in place by the clamping mechanism 112. As illustrated in
Once plasma is removed from container 114, the same procedure as described above for syringe 120 and cannula 170 is repeated with the 5 ml concentrate extraction syringe 122 and cannula 172.
Thus, in the examples illustrated in
Additional features of the above embodiment, as illustrated in
a) The collection syringe (container) 114 is centrifuged ‘luer tip up’ and an O-ring or gasket is used to keep the syringe liquid-tight during centrifugation. If a syringe is centrifuged with the luer tip facing up, no cap or a vented cap for the luer tip should be used; otherwise the syringe distorts and leaks.
b) The use of luer connectors, injection ports, vented caps and a manifold 160 as a means to both 1) vent the collection syringe (container) and 2) insert a cannula into the syringe through the luer tip to extract fluid.
The collection syringe 114 is vented and fluid removed by employing the following novel features (see, e.g.,
a) The connection between the extraction cannulas 120, 122 and the extraction syringes 170, 172 is air tight.
b) The seal around the upper injection port 162 and the cannula (170, 172) that has pierced it is also air tight.
c) The column of the manifold 160 is air tight with the exception of the air vent 164 at right angle to the syringes.
d) The connection 166 to the collection syringe 114 is air tight.
The collection syringe 114 is vented during the retrieval of the one or more target fractions. A target fraction is removed via a cannula using the negative pressure of an extraction syringe. The air vent 164 that used to accomplish this is not part of either syringe but is connected to both syringes, e.g., via the manifold 160. The luer tip 130 of the collection syringe 114 is used as the extraction port.
An alternative extraction process using a syringe PRP (Platelet Rich Plasma) system will be described. In the embodiment described above, e.g., in reference to
An alternative to removing the plasma with a cannula after clamping the funnel 116 in place is to attatch a syringe 120 to the upper luer 130 of collection syringe 114 using a standard fem/fem luer connection and remove the plasma directly, without a cannula, by using the vacuum pressure of the two connected syringes (
The target cells will not re-mix because the walls of the funnel 116 prevent fluid turbulance or interference from the inner wall of the syringe barrel. In addition, the center hole of the funnel 116 is sized such that at 1G force, surface tension prevents fluid passage from below the funnel into the funnel. The PRP extraction syringe 122 is connected to a cannula 172 that fits through an upper injection port 162, connected to a manifold 116 that has a side air vent 164 and a lower luer connection 166. The cannula 172 can be shorter for this extraction process as the float 116 is at the top of the collection syringe 114. Thus, the collection syringe 114 is not vented during loading of the specimen but is vented during extraction the target fraction inside the funnel.
After removal of the plasma and after the PRP extraction syringe 122 and cannula 172 are connected to the collection syringe 114, the user pushes the extraction cannula 172 into the collection syringe. Since the insert 116 ends at the top of the syringe in this embodiment, the cannula 172 can be a set length, such that the cannula will advance the proper distance until it is approximately at the bottom of the funnel 116. As described above in reference to
Returning to
Method 1: The First Method Involves the Following Procedures:
a) After centrifugation, pinch the funnel in place by applying a clamp on the outside of the tube (e.g., container having a fixed bottom). The pressure on the tube causes the tube to flex. This then causes the inside wall of the tube to pinch against the outside wall of the funnel.
b) Once the funnel is secured in place, a double needle apparatus (inner cannula within outer cannula) is inserted through a center port. The blunt tip of the needle mates with the center hole of the funnel blocking off fluid below the funnel from fluid above the funnel.
c) The upper access hole (side port) of the double needle extracts all fluid that resides above the top of the funnel
d) The lower access hole (side port) of the double needle extracts all fluid that resides inside the funnel.
Method 2: The Second Method Involves the Following Procedure:
a) The fluid is loaded into a syringe (e.g., a container having a movable bottom or plunger) containing the funnel; the syringe can have no plunger handle or can have a removable plunger handle
b) The syringe has a center luer connection and a side luer connection, both of which are closed.
c) In one example, the center luer connection on the underside, inside the barrel of the syringe, has connected to it a blunt needle with side ports
d) After centrifugation, the target cells reside inside the funnel
e) Upper fluid (plasma) is removed via a plasma-syringe through the side luer connection. Because the system is a syringe, it is under vacuum pressure; consequently, as fluid is removed, the funnel and plunger move up.
f) Once the majority of the fluid has been removed, the blunt end of the needle meets the center hole of the rising float (funnel) and effectively seals all fluid above the float from fluid below the float. The port retrieving the plasma also mates with the float simultaneously so that no further fluid can be removed from the side port once the blunt needle impales the rising float.
g) The plasma syringe is removed and a vented cap added which now makes the system not under vacuum pressure.
h) Another syringe is connected to the center port and the target cells are removed.
Thus, of the two methods for removing the target cells from the funnel described above, the first involves freezing (e.g., clamping) the float in place and moving the needle assembly into the funnel to get the cells, while the second method involves using the vacuum pressure from the syringe to move the funnel up so that it impales itself into the fixed blunt needle assembly under the cap of the syringe. Please refer to WO 2010/138895 A2, e.g., FIGS. 13-33 and associated text, for a more complete description of the two methods described above.
Described below in reference to
After centrifugation, the bulk of the upper funnel 210 is floating above the plasma 2000, as illustrated in
Beneath the upper funnel 210 is a second funnel 216 that has a density of about 1.06, which is a higher density than the upper funnel and which allows the lower funnel 216 to float at the intermediate zone between red cells 2004 and plasma 2000. After centrifugation, most of the platelets and white cells from blood or marrow reside within the lower funnel, as illustrated in
After centrifugation, the user can attach a syringe to the upper port 228 and begin to retrieve plasma first. The upper funnel 210 sinks as fluid in the chamber of tube 214 is removed but initially remains floating on the top of the fluid. After the desired amount of plasma has been removed, the user can switch syringes and begin to remove the remaining fluid above the lower funnel 216 and the contents of what is inside the lower funnel.
As additional fluid is removed, the upper funnel 210 continues to sink until it hits the lower funnel 216. The angle of the upper funnel 210 is steeper than the angle of the lower funnel 216 so that the upper funnel fits into the lower funnel. The upper funnel 210 can have bottom stabilization fins 225, so that the point of the upper funnel stops a certain distance below the bottom of the lower funnel 216. This can also be accomplished by making the upper funnel 210 a certain height so that the upper stabilization fins 215 contact the upper surface of the lower funnel 216. Additionally, a clamp 212 can be added after centrifugation that pinches the sidewall of the collection tube, such that the sidewall deforms and pinches against an outer surface of the lower funnel. In this way, the lower funnel 216 can be clamped in place and does not move as the upper funnel 210 mates with it during extraction of fluid. This allows the user to only withdraw the contents of the lower funnel 216 but not any fluid which is contained beneath the lower funnel.
As illustrated in
The teachings of all patents, published applications and references cited herein are incorporated by reference in their entirety.
While this invention has been particularly shown and described with references to example embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims. For example, the double plunger collection syringe described herein may be used to separate fluid components other than those described herein and may be used in applications other than those described herein. Further, the example manifold disclosed herein, including the luer connection ports and micron vent, may be used to transfer fluids in a sterile manner in other applications, and may be used with syringes, cannulas, and containers other than those described herein. Further, inserts other than those illustrated and described herein may be used in combination with containers, cannulas and syringes to separate components of a fluid. For example, inserts need not have a density as described herein.
This application is a continuation of U.S. application Ser. No. 16/835,053, filed on Mar. 30, 2020, which is a divisional of U.S. application Ser. No. 14/764,115, filed Jul. 28, 2015, now U.S. Pat. No. 10,603,665, which is the U.S. National Stage of International Application No. PCT/US2014/013636, filed on Jan. 29, 2014, published in English, which claims the benefit of U.S. Provisional Application No. 61/757,993, filed on Jan. 29, 2013 and U.S. Provisional Application No. 61/897,587, filed on Oct. 30, 2013. The entire teachings of the above applications are incorporated herein by reference.
| Number | Name | Date | Kind |
|---|---|---|---|
| 3814248 | Lawhead | Jun 1974 | A |
| 3957654 | Ayers | May 1976 | A |
| 4001122 | Griffin | Apr 1977 | A |
| 4152270 | Cornell | May 1979 | A |
| 4818418 | Saunders | Apr 1989 | A |
| 4844818 | Smith | Jul 1989 | A |
| 4917801 | Luderer et al. | Apr 1990 | A |
| 4939087 | Van Wie et al. | Jul 1990 | A |
| 5030341 | McEwen et al. | Jul 1991 | A |
| 5053134 | Luderer et al. | Oct 1991 | A |
| 5236604 | Fiehler | Aug 1993 | A |
| 5269927 | Fiehler | Dec 1993 | A |
| 5271852 | Luoma | Dec 1993 | A |
| 5308506 | McEwen et al. | May 1994 | A |
| 5474687 | Van Vlasselaer | Dec 1995 | A |
| 5489386 | Saunders | Feb 1996 | A |
| 5577513 | Van Vlasselaer | Nov 1996 | A |
| 5601711 | Sklar | Feb 1997 | A |
| 5641622 | Lake et al. | Jun 1997 | A |
| 5739033 | Soon-Shiong | Apr 1998 | A |
| 5840502 | Van Vlasselaer | Nov 1998 | A |
| 6051146 | Green et al. | Apr 2000 | A |
| 6123655 | Fell | Sep 2000 | A |
| 6221315 | Giesler et al. | Apr 2001 | B1 |
| 6410334 | Schmolz | Jun 2002 | B1 |
| 6516953 | DiCesare et al. | Feb 2003 | B1 |
| 6596179 | Giesler et al. | Jul 2003 | B2 |
| 6733433 | Fell | May 2004 | B1 |
| 7077273 | Ellsworth et al. | Jul 2006 | B2 |
| 7179391 | Leach et al. | Feb 2007 | B2 |
| 7223346 | Dorian et al. | May 2007 | B2 |
| 7316932 | Woodside | Jan 2008 | B2 |
| 7374678 | Leach et al. | May 2008 | B2 |
| 7445125 | Ellsworth et al. | Nov 2008 | B2 |
| 7547272 | Ellsworth et al. | Jun 2009 | B2 |
| 7514075 | Hedrick et al. | Jul 2009 | B2 |
| 7585670 | Hedrick et al. | Sep 2009 | B2 |
| 7595043 | Hedrick et al. | Sep 2009 | B2 |
| 7598089 | Collins | Oct 2009 | B2 |
| 7780860 | Higgens et al. | Aug 2010 | B2 |
| 7832566 | Leach et al. | Nov 2010 | B2 |
| 7845499 | Higgens et al. | Dec 2010 | B2 |
| 7914689 | Higgens et al. | Mar 2011 | B2 |
| 7954646 | Leach et al. | Jun 2011 | B2 |
| 7992725 | Leach et al. | Aug 2011 | B2 |
| 8048321 | Leach et al. | Nov 2011 | B2 |
| 8048678 | Duffy, Jr. et al. | Nov 2011 | B2 |
| 8062534 | Higgens et al. | Nov 2011 | B2 |
| RE43547 | Ellsworth et al. | Jul 2012 | E |
| 8241592 | Duffy et al. | Aug 2012 | B2 |
| 9272083 | Duffy | Mar 2016 | B2 |
| 9421319 | Hwang | Aug 2016 | B2 |
| 10005081 | Duffy et al. | Jun 2018 | B2 |
| 10603665 | Levine et al. | Mar 2020 | B2 |
| 20020006360 | Neal et al. | Jan 2002 | A1 |
| 20030205538 | Dorian et al. | Nov 2003 | A1 |
| 20050109716 | Leach et al. | May 2005 | A1 |
| 20060178644 | Reynolds | Aug 2006 | A1 |
| 20060273049 | Leach et al. | Dec 2006 | A1 |
| 20060273050 | Higgens et al. | Dec 2006 | A1 |
| 20060278588 | Woodell-May | Dec 2006 | A1 |
| 20070088252 | Pestotnik | Apr 2007 | A1 |
| 20070131612 | Duffy et al. | Jun 2007 | A1 |
| 20070265558 | Kleinbloeseum et al. | Nov 2007 | A1 |
| 20080171951 | Fell | Jul 2008 | A1 |
| 20090186065 | Tillman et al. | Jul 2009 | A1 |
| 20090283524 | Ellsworth et al. | Nov 2009 | A1 |
| 20100140182 | Chapman et al. | Jun 2010 | A1 |
| 20100256595 | Leach et al. | Oct 2010 | A1 |
| 20110056893 | Leach et al. | Mar 2011 | A1 |
| 20110086426 | Freund | Apr 2011 | A1 |
| 20110168193 | Leach et al. | Jul 2011 | A1 |
| 20120015796 | Leach et al. | Jan 2012 | A1 |
| 20120082652 | Sungun et al. | Apr 2012 | A1 |
| 20120129676 | Duffy et al. | May 2012 | A1 |
| 20130079212 | Ellsworth et al. | Mar 2013 | A1 |
| 20140148325 | Jo et al. | May 2014 | A1 |
| 20140227732 | Saqi et al. | Aug 2014 | A1 |
| 20150064687 | Nemirovsky | Mar 2015 | A1 |
| 20160008808 | Levine et al. | Jan 2016 | A1 |
| 20160158716 | Vos et al. | Jun 2016 | A1 |
| 20160263571 | Duffy et al. | Sep 2016 | A1 |
| 20200324285 | Levine et al. | Oct 2020 | A1 |
| 20200360921 | McGillicuddy | Nov 2020 | A1 |
| Number | Date | Country |
|---|---|---|
| 103657756 | Mar 2014 | CN |
| 0493838 | Jul 1992 | EP |
| 1289618 | Apr 2001 | EP |
| 2407245 | Jan 2012 | EP |
| 200183068 | Nov 2001 | WO |
| 2003099412 | Dec 2003 | WO |
| 2007050986 | Mar 2007 | WO |
| 2010058806 | May 2010 | WO |
| 2010138895 | Dec 2010 | WO |
| WO-2010138895 | Dec 2010 | WO |
| WO-2014036945 | Mar 2014 | WO |
| 2014120797 | Aug 2014 | WO |
| 2019108937 | Jun 2019 | WO |
| Entry |
|---|
| Notice of Allowance, U.S. Appl. No. 15/041,588, entitled, “Apparatus and Methods for Aspirating and Separating Components of Difference Densities from a Physiological Fluid Containing Cells,” dated Mar. 1, 2018. |
| Non-Final Office Action, U.S. Appl. No. 15/041,588, entitled, “Apparatus and Methods for Aspirating and Separating Components of Difference Densities from a Pysiological Fluid Containing Cells,” dated Mar. 1, 2018, date of mailing: Sep. 26, 2017. |
| “GPS III Platelet Separation System”, Biomet Biologies, (8 pages), Jul. 15, 2007. |
| “RES-Q60 BMC Bone marrow concentrate”, retrieved on May 4, 2010 from Thermogenesis Website URL: http://www.thermogenesis.com/CMSFiles/Pdf/Literature/resqbmc.pdf. |
| Final Office Action, U.S. Appl. No. 13/322,616, entitled “Apparatus And Methods For Aspirating And Separating Components Of Different Densities From A Physiological Fluid Containing Cells,” dated Jul. 30, 2015. |
| International Preliminary Report on Patentability from International Application No. PCT/US2010/036696, entitled “Apparatus And Methods For Aspirating And Separating Components Of Different Densities From A Physiological Fluid Containing Cells”, dated Dec. 8, 2011. |
| International Preliminary Report on Patentability from International Application No. PCT/US2014/013636, entitled “Cell Concentration Devices And Methods”, dated Aug. 4, 2015. |
| International Search Report and Written Opinion of the International Searching Authority from International Application No. PCT/US2014/013636, entitled “Cell Concentration Devices and Methods,” dated Jun. 4, 2014. |
| Joupperi, et al., “Isolation of Bone Marrow-Derived Stem Cells Using Density-Gradient Separation”, Exp. Hematol, 35(2): 335-341 (Feb. 2007). |
| Zhang, et al., “Isolating and Culturing Rat Marrow Mesenchymal Stem Cells and Studying their Phenotypical and Functional Properties”, Sichuan Da Xue Xue Bao Yi Xue Ban. 34(4): 738-741 (Oct. 2003). Abstract Only. |
| Office Action, U.S. Appl. No. 13/322,616, entitled “Apparatus And Methods For Aspirating And Separating Components Of Different Densities From A Physiological Fluid Containing Cells,” dated Apr. 3, 2015. |
| Notice of Allowance, U.S. Appl. No. 13/322,616, entitled, “Apparatus and Methods for Aspirating and Separating Components of Different Densities from a Physiological Fluid Containing Cells,” dated Oct. 27, 2015. |
| International Search Report and the Written Opinion of the International Searching Authority from International Application No. PCT/US2010/036696, entitled “Apparatus And Methods For Aspirating And Separating Components Of Different Densities From A Physiological Fluid Containing Cells,” dated Aug. 18, 2011. |
| “RES-Q60 BMC Point-of-Care Automated Cell Capturing System”, retrieved Apr. 2010 from TotipotentSC Corporate Website URL: http://totipotentsc.com/products/RES-Q60M_V2.pdf. |
| NonFinal Office Action, U.S. Appl. No. 14/764,115 entitled: “Cell Concentration Devices And Methods,” dated Mar. 9, 2018. |
| Duregger et al., “Influence of storage conditions on the release of growth factors in platelet-rich blood derivatives,” Current Directions in Biomedical Engineering, vol. 2, No. 1, pp. 311-314, (2016). |
| International Search Report and the Written Opinion of the International Searching Authority from International Application No. PCT/US2018/063317, entitled “Apparatus And Method For Centrifuging A Biologic,” dated Feb. 4, 2019. |
| International Preliminary Report on Patentability for International Application No. PCT/2018/063317, entitled “Apparatus and Method for Centrifuging a Biologic,” dated Jun. 2, 2020. |
| Non-Final Office Action for U.S. Appl. No. 16/767,863, entitled “Apparatus and Method for Centrifuging a Biologic,” dated Mar. 17, 2022. |
| Non-Final Office Action for U.S. Appl. No. 16/835,053, entitled “Cell Concentration Devices and Methods That Include an Insert Defining a Lumen and a Cannula Assembly,” dated Nov. 2, 2022. |
| Number | Date | Country | |
|---|---|---|---|
| 20220226812 A1 | Jul 2022 | US |
| Number | Date | Country | |
|---|---|---|---|
| 61757993 | Jan 2013 | US | |
| 61897587 | Oct 2013 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 14764115 | US | |
| Child | 16835053 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 16835053 | Mar 2020 | US |
| Child | 17715222 | US |
