The application relates to multi-layered cell culture vessels and, more particularly, to closed-volume multi-layered cell culture vessels which are accessed through a port of limited diameter.
Multi-layered cell culture vessels are known in the prior art, including that disclosed in U.S. Published Patent Application No. 2011/0020923, to the Assignee herein. U.S. Published Patent Application No. 2011/0020923 is incorporated by reference herein. Multi-layered cell culture vessels provide a higher surface to volume ratio for a cell culture for a given footprint as compared to non-layered, standard open volume cell culture flasks and similar structures. Improvements in the ability to handle multi-layered cell culture vessels, as well as, in the ability to prepare and remove cultured cells in and from, respectively, multi-layered cell culture vessels are desirable.
In one aspect, a multi-layered cell culture vessel is provided herein which includes a body enclosing a closed volume. The body includes at least one insert layer located within the closed volume to separate portions thereof, and at least one port allowing communication with the closed volume from exteriorly of the body. At least one grip protrudes from the body configured to be grippingly engaged by a user in transporting the multi-layered cell culture vessel. Advantageously, an external grip is provided herein which permits for better handling than with traditional multi-layered cell culture vessels.
Additional features are disclosed herein including the use of a side port which is disposed transversely to a longitudinal axis of the body, including being disposed to intersect a gap located adjacent to each of the insert layers. This arrangement allows for the preparation and removal of cell culture medium without obstruction from the insert layers. Additionally, the end of the body may be formed with tapered portions that channel flow towards an end port thereby allowing for a relatively well-defined stream to be poured from the device as needed.
These and other features of the invention will be better understood through a study of the following detailed description and accompanying drawings.
With reference to the Figures, a multi-layered cell culture vessel is shown and generally designated with the reference numeral 10. The vessel 10 includes a body 12 which encloses a closed volume 14. At least one insert layer 16 is located within the closed volume 14 to separate portions thereof. The insert layers 16 provide additional surface area within the closed volume 14 for cell culturing. As will be appreciated by those skilled in the art, the insert layers 16 may be of various configurations, and provided in various quantities, including being in a stacked configuration, as best shown in
At least one port 24 is provided with the body 12 to allow communication with the closed volume 14 from exteriorly of the body 12. The port 24 may be in the form of one or more side ports 24A and/or in the form of one or more end ports 24B.
To facilitate handling of the vessel 10, at least one grip 26 may be provided which protrudes from the body 12 and is configured to be grippingly engaged by a user in transporting the vessel 10. As will be appreciated by those skilled in the art, the grip 26 may be formed with various configurations. In a preferred arrangement, the grip 26 is arcuately formed with ends 28 being fixed to the body 12 and a space 30 being defined between the grip 26 and the body 12 configured to receive a hand of a user.
The body 12 preferably defines a generally rectangular footprint as viewed in a first direction with a longitudinal dimension L which is preferably greater than a transverse dimension W. Accordingly, the body 12 includes first and second sides 32, 34 which are generally parallel to the longitudinal dimension L, and first and second ends 36, 38 which are generally parallel to the transverse dimension W. Preferably, the grip 26 is located on the first side 32 with at least one of the end ports 24B being located on the second end 38. This allows for the grip 26 to be used to conveniently pour cell media from the vessel 10 via the end port(s) 24B.
At least one of the side ports 24A may protrude from the body 12. The side port 24A may be generally tubular and extend along a secondary longitudinal axis R which is disposed generally parallel to the transverse dimension W. As known in the art, a cap, or other sealing member, may be mounted to the side port 24A to cause selective sealing thereof. The cap may be vented or solid to permit or prevent gas transmission therethrough, as needed. In addition, tubing or other instrumentation may be passed through the cap. The side port 24A may protrude from the body 12 in a direction generally parallel to the direction of the grip 26. It is preferred that the grip 26 protrude from the body 12 no further than the side port 24A with a cap mounted thereto.
The use of the one or more side ports 24A and/or one or more of the end ports 24B permits for gas exchange through multiple directions within the closed volume 14. The ports 24A, 24B may be in varying combinations provided with vented or closed caps to control gas transmission and flow.
In addition, the insert layers 16 may be formed to not extend fully across the body 12. With reference to
Also, the one or more side ports 24A may be used to prepare a cell culture within the vessel 10 with media and/or reagents being introduced into the closed volume 14, either by pouring, pipette or tubing, through the one or more side ports 24A. The filling can be accomplished either by gravity filling or pump-feeding. It is preferred to fill the vessel 10 with any media or reagents with the body 12 resting on the second side 34 so that the one or more side ports 24A protrude upwardly. This presents an equilibration position of the vessel 10 where introduced media and reagents are caused to collect in the closed volume 14 as a volume adjacent to the second side 34. In this position, the introduced media and reagents equilibrate across the insert layers 16. Once filled, the grip 26 may be used to transport the vessel 10 to a desired location, such as an incubator. Once at the desired location, and possibly using the grip 26, the body 12 is re-oriented to rest on the bottom 22. With re-orientation, the media partitions amongst the insert layers 16 in even amounts. Content may be removed through one or more of the end ports 24B by pouring or using tubing with gravity or pump extraction. The vessel 10 may be stood on an inclined support to assist in draining.
In a preferred arrangement, the second end 38 may be formed to channel flow towards the one or more end ports 24B. The second end 38 may include first and second portions 42, 44 which are disposed transversely to the secondary longitudinal axis R. In addition, the second end 38 may include a third portion 46 which extends between the first and second portions 42, 44. The third portion 46 may be generally planar and disposed generally parallel to the secondary longitudinal axis R. Preferably, any utilized end ports 24B are secured to the second end 38 at the third portion 46. The first and second portions 42, 44 are disposed to converge towards the third portion 46 so as to provide a funneling effect to any end port 24B located on the third portion 46. In addition, it is preferred that any utilized side port 24A be secured to the first portion 42. With this arrangement, an enlarged space is created below the side port 24A with portions of the second end 38 being recessed. During media introduction, this advantageously minimizes the extent of cell adherence to portions of the body 12 adjacent to the side port 24A. In addition, this space may act as an air reservoir from which air may be distributed amongst the insert layers 16.
Preferably, the top 20 and the bottom 22 are formed generally planar, and more preferably parallel, to permit level stacking of a plurality of the vessels 10. Ribs or other features may be included on one or both of the top 20 and the bottom 22 to facilitate stacking alignment and stability. Also, a pouch or other containment member may be secured or otherwise supported by the top 20 during use with substance being introduced therefrom via tubing through either of both of the side ports 24A and the end ports 24B.
As will be appreciated by those skilled in the art, the various features discussed herein may be used singularly or in various combinations.
This application claims priority to U.S. Provisional Patent Application No. 61/719,156, filed on Oct. 26, 2012, the entire contents of which are incorporated by reference herein.
Number | Date | Country | |
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61719156 | Oct 2012 | US |