Microcavity cell culture vessel with inclined surface having diverter - Patent Application 20070122997

Abstract

The cell culture vessel includes a vessel body having a first end wall, a second end wall opposite the first end wall, and a pair of side walls extending between the first and second end walls. The first end wall, the second end wall, and the pair of side walls define the outer periphery of a cell culture chamber. An inner filling wall extends inwardly into the cell culture chamber from the first end wall and the pair of side walls. The inner filling wall has a sloped inner surface, the sloped inner surface having an outer edge at the first end wall that is angularly offset from an inner terminal edge of the sloped inner surface. A diverter extends along the sloped inner surface between the pair of side walls and at least partially transverse to the filling direction. The diverter provides a passageway between the diverter and the pair of side walls through which liquid flows by the diverter during a filling operation.

Description

Related Applications

[0001] This application claims the benefit of priority under 35 U.S.C. § 119 to U.S. Provisional Patent Application No. 63 / 411,210, filed September 29, 2022, the contents of which are relied upon and incorporated herein by reference in their entirety. [Technical Field]

[0002] The present specification relates generally to cell culture vessels used for growing cells, and more particularly to microcavity cell culture vessels having diverters for containing and controlling liquid flowing down the inclined sides of the cell culture vessel. [Background technology]

[0003] In general, three-dimensional (3D) cell cultures can simulate the environment of natural tissues and organs better than two-dimensional (2D) cell cultures grown in monolayers. Cells grown in 3D cell cultures can attach to other deposited cells within the three-dimensional environment to form spheroids or cell aggregates, thereby creating more natural cell-to-cell interactions than 2D cells grown in monolayers. This cell arrangement results in a flexible configuration similar to that of natural tissues. Providing an accurate representation of the tissue microenvironment is desirable. Because 3D cultures more closely resemble the environment in which developed drugs will ultimately be applied, developing therapies in 3D cultures instead of 2D cultures is desirable for increased precision in experimental research to develop therapeutics for disease.

[0004] However, the formed spheroids or cell aggregates are easily damaged, for example, during transport of the cell culture vessel or during medium changes. Transporting 3D cell culture vessels typically involves unintentional movement of the liquid contained within, creating turbulence within the vessel. Turbulence can also occur when adding or removing liquid medium from the vessel during the 3D cell culture process. Because 3D cell culture cells are not attached to any surface of the vessel, turbulence can cause spheroids or cell aggregates to slosh in or detach from their respective microcavities within the 3D culture vessel in which they were formed. When spheroids detach from their respective microcavities, they may attach to other spheroids, potentially resulting in loss of spheroid uniformity and size within the vessel. Therefore, it is necessary to stabilize the liquid movement within the 3D spheroid culture vessel. Summary of the Invention

[0005] According to one embodiment, a cell culture vessel includes a vessel body having a first end wall, a second end wall opposite the first end wall, and a pair of side walls extending between the first and second end walls. The first end wall, the second end wall, and the pair of side walls define the outer periphery of a cell culture chamber. An inner fill wall extends inwardly into the cell culture chamber from the first end wall and the pair of side walls. The inner fill wall has a sloped inner surface, the sloped inner surface having an outer edge at the first end wall that is angularly offset from an inner terminal edge of the sloped inner surface. A diverter extends along the sloped inner surface between the pair of side walls and at least partially transverse to the fill direction. The diverter provides a passageway between the diverter and the pair of side walls through which liquid flows by the diverter during a fill operation.

[0006] According to another embodiment, a method of forming a cell culture vessel is provided. The method includes forming the vessel body as a single monolithic piece, such that the vessel body has a first end wall, a second end wall opposite the first end wall, and a pair of side walls extending between the first and second end walls. The first end wall, the second end wall, and the pair of side walls define an outer periphery of a cell culture chamber. An inner filling wall extends inwardly into the cell culture chamber from the first end wall and the pair of side walls. The inner filling wall has a sloped inner surface, the sloped inner surface having an outer edge at the first end wall that is angularly offset from an inner terminal edge of the sloped inner surface. A diverter extends along the sloped inner surface between the pair of side walls and at least partially transverse to the filling direction. The diverter provides a passageway between the diverter and the pair of side walls through which liquid flows during a filling operation. A protective carrier is formed separately from the vessel body. The protective carrier includes a plate sized and configured to cover the cell culture surface of the vessel body.

[0007] Additional features and advantages of the cell culture vessels described herein are set forth in the detailed description which follows, and in part will be readily apparent to those skilled in the art from that description, or may be learned by practicing the embodiments described herein, including the following detailed description, claims, and accompanying drawings.

[0008] It is to be understood that both the foregoing general description and the following detailed description are intended to describe various embodiments and provide an overview or framework for understanding the nature and features of the claimed subject matter. The accompanying drawings are included to provide a further understanding of the various embodiments, and are incorporated into and constitute a part of this specification. The drawings illustrate various embodiments described herein and, together with the description, serve to explain the principles and operation of the claimed subject matter. [Brief explanation of the drawings]

[0009] [Figure 1] FIG. 1 is a perspective view of a cell culture vessel including a vessel body having a diverter and a protective carrier according to one or more embodiments shown and described herein. [Figure 2] 2 is a side cross-sectional view of the container body of FIG. 1 in an isolated state according to one or more embodiments shown and described herein. [Figure 3] 3 is a cross-sectional end view of the container body taken along line 3-3 of FIG. 2 according to one or more embodiments shown and described herein. [Figure 4] 2 is a top perspective view of the protective carrier of FIG. 1 in an isolated state according to one or more embodiments shown and described herein. [Figure 5] 5 is a bottom perspective view of the protective carrier of FIG. 4 according to one or more embodiments shown and described herein. [Figure 6] 2 is a side perspective view of a portion of a cell culture substrate used in the cell culture vessel of FIG. 1 according to one or more embodiments shown and described herein. [Figure 7] 2 illustrates the cell culture vessel of FIG. 1 in use, according to one or more embodiments shown and described herein. DETAILED DESCRIPTION OF THE INVENTION

[0010] Reference will now be made in detail to various embodiments of cell culture vessels having diverters disposed therein, examples of which are illustrated in the accompanying drawings. Wherever possible, the same reference numbers will be used throughout the drawings to refer to the same or like parts. Directional terms used herein, such as up, down, right, left, front, back, upper, bottom, distal, and proximal, are used only with reference to the figures depicted and are not intended to imply absolute directions.

[0011] Ranges can be expressed herein as from "about" one particular value, and / or to "about" another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.

[0012] Unless otherwise expressly stated, it is not intended that any method described herein be construed as requiring its steps to be performed in a particular order, or that any particular direction be required by any apparatus. Thus, if a method claim does not actually recite the order in which its steps must be followed, or any apparatus claim does not actually recite an order or direction for individual components, or if the claim or specification does not otherwise specifically state that the steps are limited to a particular order or recite a particular order or direction for the apparatus components, then no particular order or direction is intended to be inferred in any respect. This applies to all possible implicit grounds of interpretation, including logical considerations regarding the arrangement of steps, operational flow, component order, or component direction; the apparent meaning derived from grammatical construction or punctuation; and the number or type of embodiments described herein.

[0013] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a" element includes aspects having two or more such elements unless the context clearly dictates otherwise.

[0014] Liquid in a cell culture vessel with a large open volume moves freely and somewhat uncontrolled across the open area. When this cell culture vessel is moved or transported, the movement can cause turbulence in the liquid, resulting in "sloshing" and potential spillage. Unlike cells cultured in two-dimensional (2D) culture, which adhere to the vessel's culture surface, cells such as spheroids in 3D culture may not be attached to the culture surface and may be prone to sloshing, potentially causing the cells to detach from the microcavities in which they are cultured. Such turbulence and detachment of spheroids from the microcavities can result in spheroid loss. Detached spheroids may eventually settle in the microcavities with other spheroids, resulting in undesirable uneven spheroid size and loss of spheroid uniformity.

[0015] Furthermore, filling the microcavities with medium without trapping air and exchanging the medium after spheroid formation requires some skill. Trapped air can prevent cells from settling into the microcavities. Furthermore, because cells grow in 3D within cell culture vessels, the vessels may contain more cells than would normally be contained within a 2D cell growth surface area. This increased cell number may require more frequent medium changes or require the maintenance of a larger volume of fluid to meet the metabolic demands of the cells.

[0016] Embodiments described herein relate to a cell culture container including a filling structure that can be used to control the filling of the cell culture container with liquid medium. The filling structure can also control the movement of medium after the cell culture container is filled. The cell culture container includes a container body including a first end wall, a second end wall opposite the first end wall, and a pair of side walls extending between the first and second end walls. The first end wall, the second end wall, and the pair of side walls form the outer periphery of a cell culture chamber when a floor and a lid are connected to the container body. The container body includes a neck extending outward from the first end wall in a filling direction to an opening. The neck includes a sloped filling surface, and the sloped filling surface has an outer edge at the opening that is angularly offset from an inner edge of the sloped filling surface at the first end wall. An inner filling wall extends inward from the first end wall and the pair of side walls into the cell culture chamber. The inner filling wall has a sloped inner surface, and the sloped inner surface has an outer edge at the first end wall that is angularly offset from an inner terminal edge of the sloped inner surface. The diverter extends along an inclined inner surface at least partially transverse to the filling direction between the pair of side walls, and has opposing ends that terminate inwardly of the pair of side walls to define a passageway through which liquid flows by the diverter during the filling operation.

[0017] 1 and 2, one embodiment of a cell culture vessel 10 (FIG. 1) is a microcavity flask that includes a vessel body 12 (FIG. 2). The vessel body 12 includes a first end wall 14 and a second end wall 16 opposite the first end wall 14. Side walls 18 and 20 extend between the first end wall 14 and the second end wall 16. In some embodiments, the first and second end walls 14 and 16, or portions thereof, are substantially parallel, and a pair of side walls 18 and 20, or portions thereof, are substantially parallel to form a somewhat rectangular perimeter 22 of a cell culture chamber 24. While a rectangular perimeter 22 is illustrated, any suitable perimeter shape can be used.

[0018] Vessel body 12 includes a neck 26 that extends outward from first end wall 14 in the fill direction to an opening 28. Opening 28 provides an inlet point for liquid medium to enter cell culture vessel 10 and flow into cell culture chamber 24 during a fill operation. Opening 28 also provides an outlet point for liquid medium to exit cell culture chamber 24 during a drain operation. Neck 26 can optionally be configured to mate with a cap 27, for example, using a threaded, snap-on, or any suitable releasable connection.

[0019] Neck 26 includes a sloped fill surface 30 forming a portion of neck 26, with an outer edge 32 at opening 28 that is angularly offset from an inner edge 34 of sloped fill surface 30 at first end wall 14. Inner fill wall 36 extends inwardly into cell culture chamber 24 from first end wall 14 and pair of side walls 18 and 20. Inner fill wall 36 has a sloped inner surface 38 at first end wall 14 that has an outer edge 40 that is angularly offset from an inner terminal edge 42 of sloped inner surface 38. In some embodiments, sloped inner surface 38 is an extension of sloped fill surface 30 of neck 26, such that when cell culture vessel 10 is in the horizontal incubation position, both sloped inner surface 38 and sloped fill surface 30 intersect and extend at the same angle θ (e.g., 10 degrees or more, 15 degrees or more, 20 degrees or more, 10-20 degrees, 15-20 degrees) relative to the horizontal plane, as shown in FIGS.

[0020] Cell culture vessel 10 further includes a lid 44 that covers a viewing surface 46 of vessel body 12. Lid 44 includes a lip 47 that engages with edges 48 of first and second end walls 14, 16 and side walls 18, 20 to provide an interlocking connection. Lid 44 may be formed of a transparent material, such as a clear or translucent plastic, to permit visual inspection of the contents within cell culture chamber 24.

[0021] Cell culture vessel 10 may include a cell culture substrate 50 disposed along opposing cell culture surfaces 52 of vessel body 12. Cell culture substrate 50 may include a surface 110 with a plurality of microcavities 56 sized and shaped to accommodate at least one cell or spheroid therein. Cell culture surface 110 is thus a cell culture area configured to promote the growth and development of cells within cell culture chamber 24.

[0022] Referring to FIG. 2 , the vessel body 12 forms a frame 58 on the cell culture surface 52 of the vessel body 12 that is sized and configured to accommodate the cell culture substrate 50 ( FIG. 1 ). In particular, the frame 58 may be formed at one edge 61 thereof between a ledge 62 extending outward from the inner filler wall 36, away from the cell culture chamber 24, and an inner terminal edge 42 of the inner filler wall 36 that is offset below the inclined inner surface 38. An opposite edge 63 of the frame 58 may be formed between another ledge 64 extending outward from the second end wall 16, away from the cell culture chamber 24, and an edge 66 of the second end wall 16. Additionally, the sides of the frame 58 may be formed along the side walls 18 and 20 that similarly include ledges 67 and 69 (see FIG. 3 ). The cell culture substrate 50 can be retained within the frame 58 during the incubation process. As can be seen, edge 68 of cell culture substrate 50 may be offset below inner terminal edge 42 of inner filler wall 36 .

[0023] 3 , a diverter 70 extends along the inclined inner surface 38. The diverter 70 extends at least partially transverse to the fill direction between the pair of side walls 18 and 20. The diverter 70 is formed as a partial height wall, with a height H1 of the diverter being less than a height H2 of the cell culture chamber 24 (e.g., 50% or less, 60% or less, 70% or less, 80% or less, 90% or less). The diverter 70 has opposing ends 72 and 74 that terminate inside the pair of side walls 18 and 20, thereby forming passageways 76 and 78 through which liquid medium flows by the diverter 70 during fill and drain operations.

[0024] The diverter 70 extends widthwise along the inclined inner surface 38. In some embodiments, the diverter 70 is integrally formed with the inner fill wall 36 as a single, monolithic piece. In other embodiments, the diverter 70 may be formed separately and connected to the inclined inner surface 38. The diverter 70 may contact and be continuous with the inclined inner surface 38 along its entire length to direct the liquid medium through the passages 76 and 78. While the diverter 70 is illustrated as having a rectangular cross-sectional shape, it may be formed having other shapes, such as curved, triangular, etc. The diverter 70 may also extend in other directions across the width of the inclined inner surface 38. For example, the diverter 70 may extend at an oblique angle to the fill direction.

[0025] The diverter 70 may extend only partially along the width W of the inclined inner surface 38 between the ends 72 and 74 to provide passages 76 and 78. For example, the diverter 70 can extend 99% or less of the width W, e.g., 95% or less, e.g., 90% or less, e.g., 85% or less, e.g., 80% or less, e.g., 75% or less, e.g., 70% or less, e.g., 65% or less, e.g., 60% or less, e.g., 55% or less, e.g., 50% or less, e.g., 45% or less, e.g., 40% or less, e.g., 35% or less, e.g., 30% or less. In some embodiments, the diverter 70 extends at least about 50% to about 90% of the width W, e.g., about 80% of the width W.

[0026] Referring to FIG. 4, protective carrier 80 is a rigid plate that can protect the gas-permeable microcavities of cell culture substrate 50 during transport and can also serve as a protective carrier for cell culture vessel 10 when used for cell culture. Protective carrier 80 can be comprised of a rigid plate 82 that is slightly larger than the footprint of cell culture vessel 10. An outer lip 84 can extend around the periphery of plate 82 and can be sized and configured to extend around inner fill wall 36 and ledges 62, 64, 67, and 69 on second end wall 16 and side walls 18 and 20. Protective carrier 80 has standoff ribs 86 at the corners. Standoff ribs 86 extend along both plate 82 and outer lip 84, providing clearance and an air gap between protective carrier 80 and cell culture substrate 50. Referring to FIG. 5, stacking grooves or ribs 88 can be provided along the underside of plate 82. Stacking grooves or ribs 88 are provided to mate with corresponding stacking ribs or grooves 90 (FIG. 1) provided on lid 44 .

[0027] Referring now to FIG. 6 , gas permeability is a property that contributes to a 3D cell culture environment. Ensuring gas permeability within the microcavities 56 of a cell culture vessel can reduce the frequency with which cell culture growth medium needs to be replaced and promote cell growth. Microcavity vessels are unique in their geometry and configuration in that they are formed from a gas-permeable substrate 50 containing micrometer-scale wells 56, also referred to as microcavities. Such microcavity vessels enjoy gas permeability due to the thickness of the microcavity substrate 50, which can be formed from very thin polystyrene material, with thicknesses ranging from about 28 μm to about 72 μm. While gas permeability can be advantageous for culturing cell aggregates, the thinness of the microcavity substrate material makes the microcavity cell culture vessel 10 susceptible to damage during shipping and use.

[0028] Each microcavity 56 may include an internal cavity with a rounded bottom 102 that is non-adhesive to cells. Thus, the microcavity containers described herein are cell culture devices that facilitate 3D cell culture by allowing cells seeded in the microcavities to self-assemble or attach to each other to form spheroids within each microcavity. The microcavities may be shallow, allowing cell culture medium to cover the spheroids, organoids, or 3D cell aggregates within the entire cavity at once, facilitating manual handling.

[0029] In one embodiment, the top surface 104 of the microcavity 56 may be recessed to a position close to the bottom of the sidewalls 18 and 20 (FIG. 3). Each individual microcavity may hold a small amount of culture medium. Each individual microcavity may have any suitable dimensions. For example, the diameter or width of each individual microcavity may be in the range of about 500 μm to about 5 mm. The depth of each individual microcavity may be in the range of about 500 μm to about 6 mm. In some embodiments, the depth of each individual microcavity may be in the range of about 500 μm to about 650 μm. In some embodiments, the depth of each individual microcavity may be about 1.6 mm. Excess culture medium may be added to the microcavity container so that the spheroids, organoids, or 3D cell aggregates do not have to rely solely on the small amount of culture medium in each individual microcavity.

[0030] In some embodiments, the microcavity substrate 50 may have a cross-sectional shape that is undulating or approximates a sinusoid. In such embodiments, the bottoms of the microcavity wells 56 are rounded (e.g., hemispherically round), the sidewalls increase in diameter from the bottom to the top of the well, and the boundaries or barriers between wells are rounded. Thus, the tops of the microcavity wells do not terminate at right angles. In some embodiments, the width of the wells is greater than the width of the barriers between adjacent wells. Such embodiments allow for a greater number of wells within a given area of ​​culture surface.

[0031] In some embodiments, the plurality of microcavities are arranged in a hexagonal close-packed pattern. In some embodiments, each microcavity has a rounded bottom. In some embodiments, each microcavity is configured so that cells cultured within the microcavity container form three-dimensional (3D) cell aggregates. In some embodiments, the inner surface of the microcavity substrate is non-adhesive to cells. In some embodiments, the inner surface of the microcavity substrate comprises a cell-non-adhesive surface coating comprising a perfluoropolymer, an olefin, a lipid, agarose, a non-ionic hydrogel, a polyether, a polyol, a polymer that inhibits cell adhesion, or a combination thereof. In some embodiments, the cell-non-adhesive surface coating comprises an ultra-low attachment (ULA) surface coating.

[0032] The microcavity substrate, microcavity container, and diverter can be formed from the same or similar materials. In some embodiments, the microcavity substrate can be molded or formed separately from the rest of the microcavity container and then joined by thermal bonding, ultrasonic welding, or any other method of plastic joining. The materials of construction of the microcavity container, microcavity substrate, and / or baffle can include "plastic" polymers, copolymers, or polymer blends. Non-limiting examples include silicone rubber, polystyrene, polypropylene, polyethylene, polyethylene terephthalate, polymethylpentene, polycarbonate, polymethyl methacrylate, styrene-ethylene-butadiene-styrene, other such polymers, or combinations thereof. In some embodiments, the microcavity substrate is formed from polydimethylsiloxane (PDMS), polymethylpentene, (poly)4-methylpentene (PMP), polyethylene (PE), polystyrene (PS), polypropylene, polyethylene terephthalate, polycarbonate, polymethyl methacrylate, styrene-ethylene-butadiene-styrene, silicone rubber or copolymer, ethylene vinyl acetate, polysulfone, polytetrafluoroethylene, poly(styrene-butadiene-styrene), or combinations thereof. Any suitable construction method can be used to form the microcavity substrate, microcavity container, and diverter, including, but not limited to, injection molding, thermoforming, 3D printing, or any other method suitable for forming plastic parts.

[0033] In some embodiments, the protective carrier 80 is formed from a polymer, metal, or glass. In some embodiments, the polymer includes polystyrene, polypropylene, polyethylene, polyethylene terephthalate, polymethylpentene, polycarbonate, polymethyl methacrylate, styrene-ethylene-butadiene-styrene, other such polymers, or combinations thereof. In some embodiments, the metal includes aluminum, stainless steel, zinc, or combinations thereof. In some embodiments, the glass includes borosilicate glass. In some embodiments, the protective carrier is formed from a renewable material. In some embodiments, the protective carrier is formed from a biodegradable material. In some embodiments, the protective carrier is opaque. In some embodiments, the protective carrier is translucent.

[0034] 1-3 , in use, a plurality of cells can be deposited within the cell culture vessel 10 such that the cell culture chamber 24 is operable to accommodate the cells within a plurality of microcavities 56 in the cell culture surface 110 of the cell culture substrate 50. Initially, it may be desirable to pre-wet the microcavity surfaces. Liquid medium can be delivered through the neck 26, along the sloped fill surface 30, and then onto the sloped inner surface 38 of the inner fill wall 36. The liquid medium can then impinge on the diverter 70, which diverts the liquid medium along its length toward the passages 76 and 78 so that the flow slows as it approaches the cell culture surface 110. The liquid medium can enter the microcavities 56 unassisted. If the liquid does not enter the microcavities, trapped air may appear as opaque areas on the cell culture surface 110. Microcavities 56 with trapped air may prevent cells in suspension from settling in those microcavities 56 after the filling process. With multiple cells contained along the cell culture substrate 50 of the cell culture surface 110, cell development is promoted by exposing the cell culture surface 110 to various nutrients and growth fluids during a liquid culture medium filling operation.

[0035] The microcavity can be deep enough to allow for gentle medium exchange, but not so deep that it is difficult to retrieve the spheroids when desired. During medium exchange, the cell culture vessel 10 can remain in the incubation position to limit spheroid loss. The inclined inner surface 38 and diverter 70 can facilitate the gradual outflow of medium from the cell culture vessel 10.

[0036] During the medium exchange step, lifting the second end wall 16 by 3–4 degrees can force the liquid medium toward the diverter 70 and the inclined inner surface 38, allowing for complete liquid removal and slowing the flow of new liquid into the cell culture vessel 10, reducing sloshing. To prevent liquid from sloshing and wetting the cap 27, it may be desirable to lift the second end wall 16 no more than 10 degrees when aspirating and dispensing liquid. Referring to FIG. 7 , a protective carrier 80 or similar article of height (e.g., 6–7 mm) can be used to lift the second end wall 16 during the medium exchange step. Notably, the ledges 67 and 69 may include a stacking recess 114 that can receive the outer lip 84 of the protective carrier 80 and function as a stop mechanism to position the cell culture vessel at the appropriate angle. To remove the spent medium, a pipette tip can be pressed against the diverter 70 and the medium can be aspirated. To exchange the medium, the pipette tip can be placed against the diverter 70 and fresh medium can be slowly added to the cell culture vessel 10. After the medium exchange is complete, the protective carrier 80 can be removed and the cell culture vessel 10 can be placed in a flat incubation position.

[0037] The above-described cell culture vessels having a diverter disposed within each cell culture chamber of the microcavity cell culture vessel, and systems for microcavity cell culture having a diverter disposed within each cell culture chamber of the microcavity cell culture vessel, reduce the amount of liquid culture medium moving within the cell culture chamber. The diverter can also improve filling by controlling liquid flow and reducing air trapped within the microcavity. The diverter can be formed (e.g., molded) with the vessel body so that the diverter and vessel body are formed as a single monolithic piece of material. The diverter can also provide a location for a pipette tip to rest against and use to remove and dispense liquid medium from and into the cell culture chamber, improving filling efficiency and reproducibility during filling and dispensing operations.

[0038] Embodiments can be described with reference to the following numbered items, with preferred features being set out in the dependent items: Item 1: A cell culture vessel comprising: a vessel body; a first end wall; a second end wall opposite the first end wall; a pair of side walls extending between the first end wall and the second end wall, wherein the first end wall, the second end wall, and the pair of side walls define an outer periphery of a cell culture chamber; an inner filling wall extending inwardly into the cell culture chamber from the first end wall and the pair of side walls, the inner filling wall having a sloped inner surface, the sloped inner surface having an outer edge at the first end wall that is angularly offset from an inner terminal edge of the sloped inner surface; and a diverter extending along the sloped inner surface between the pair of side walls in a direction at least partially transverse to the filling direction, the diverter providing a passageway between the diverter and the pair of side walls through which liquid flows by the diverter during a filling operation.

[0039] Item 2: The cell culture vessel according to item 1, wherein the entire length of the diverter is continuous with the inclined inner surface of the inner filling wall.

[0040] Item 3: The cell culture vessel according to item 2, wherein the diverter is formed as an integral, monolithic part of the inner filler wall.

[0041] Item 4: A cell culture vessel according to any one of items 1 to 3, wherein the diverter has opposing ends that terminate inside the pair of side walls to provide a passageway.

[0042] Item 5: The cell culture vessel according to any one of items 1 to 4, further comprising a lid dimensioned and configured to cover the observation surface of the vessel body.

[0043] Item 6: The cell culture vessel of any one of items 1 to 5, further comprising a protective carrier including a plate sized and configured to cover the cell culture surface of the vessel body.

[0044] Item 7: The cell culture vessel of item 6, wherein the protective carrier comprises a lip extending around the periphery of the plate sized and configured to extend along the first end wall, the end side wall, and the pair of side walls.

[0045] Item 8: The cell culture vessel of item 6 or 7, wherein the protective carrier comprises standoff ribs positioned between the plate and the vessel body and arranged and configured to provide a space between them.

[0046] Item 9: The cell culture vessel of any one of items 1 to 8, wherein the diverter extends at least about 50% to about 90% or less of the width of the inclined inner surface between both ends of the diverter.

[0047] Item 10: The cell culture vessel of any one of items 1 to 9, wherein the vessel body further comprises a neck extending outward from the first end wall in the filling direction to the opening, the neck including a sloped filling surface, the sloped filling surface having an outer edge at the opening that is angularly offset from an inner edge of the sloped filling surface at the first end wall.

[0048] Item 11: The cell culture vessel according to Item 10, wherein the inclined filling surface and the inclined inner surface intersect.

[0049] Item 12: A method for forming a cell culture vessel, the method comprising: forming the vessel body as a single monolithic piece so that the vessel body comprises: a first end wall; a second end wall opposite the first end wall; a pair of side walls extending between the first end wall and the second end wall, wherein the first end wall, the second end wall, and the pair of side walls define an outer periphery of a cell culture chamber; an inner filling wall extending inwardly into the cell culture chamber from the first end wall and the pair of side walls, the inner filling wall having a sloped inner surface, the sloped inner surface having an outer edge at the first end wall that is angularly offset from an inner terminal edge of the sloped inner surface; and a diverter extending along the sloped inner surface between the pair of side walls in a direction at least partially transverse to the filling direction, the diverter providing a passageway between the diverter and the pair of side walls through which liquid flows by the diverter during a filling operation; and separately forming a protective carrier from the vessel body, the protective carrier including a plate sized and configured to cover the cell culture surface of the vessel body.

[0050] Item 13: The method of item 12, wherein the entire length of the diverter is continuous with the sloped inner surface of the inner fill wall.

[0051] Item 14: The method of items 12 or 13, wherein the diverter has opposing ends that terminate inwardly of the pair of side walls to provide a passageway.

[0052] Item 15: The method of any one of items 12 to 14, further comprising forming a lid configured to cover the observation surface of the container body.

[0053] Item 16: The method of any one of items 12 to 15, wherein the protective carrier comprises a lip extending around the periphery of the plate sized and configured to extend along the first end wall, the end side wall, and the pair of side walls.

[0054] Item 17: The method of any one of items 12 to 16, wherein the protective carrier comprises standoff ribs positioned between the plate and the container body and arranged and configured to provide a space therebetween.

[0055] Item 18: The method of any one of items 12 to 17, wherein the diverter extends at least about 50% to no more than about 90% of the width of the inclined inner surface between opposite ends of the diverter.

[0056] Item 19: The method of any one of items 12 to 18, wherein the container body further comprises a neck extending outward from the first end wall in the filling direction to the opening, the neck including a sloped filling surface, the sloped filling surface having an outer edge at the opening that is angularly offset from an inner edge of the sloped filling surface at the first end wall.

[0057] Item 20: The method of item 19, wherein the inclined filling surface and the inclined inner surface intersect.

[0058] It will be apparent to those skilled in the art that various modifications and variations can be made to the embodiments described herein without departing from the spirit and scope of the claimed subject matter. Thus, it is intended that the present specification cover modifications and variations of the various embodiments described herein, provided such modifications and variations come within the scope of the appended claims and their equivalents.

[0059] Preferred embodiments of the present invention will be described below in detail.

[0060] Embodiment 1 A cell culture vessel, comprising: A container body is provided, the container body comprising: a first end wall; a second end wall opposite the first end wall; a pair of side walls extending between the first end wall and the second end wall, the first end wall, the second end wall and the pair of side walls defining an outer periphery of a cell culture chamber; an inner filler wall extending inwardly into the cell culture chamber from the first end wall and the pair of side walls, the inner filler wall having a beveled inner surface, the beveled inner surface having an outer edge at the first end wall that is angularly offset from an inner terminal edge of the beveled inner surface; a diverter extending along the inclined inner surface at least partially transverse to the filling direction between the pair of side walls, the diverter providing a passageway between the diverter and the pair of side walls through which liquid flows during a filling operation; A cell culture vessel comprising:

[0061] Embodiment 2 2. The cell culture vessel of embodiment 1, wherein the entire length of the diverter is continuous with the inclined inner surface of the inner filling wall.

[0062] Embodiment 3 3. The cell culture vessel of embodiment 2, wherein the diverter is formed as an integral, monolithic part of the inner filler wall.

[0063] Embodiment 4 2. The cell culture vessel of embodiment 1, wherein the diverter has opposing ends that terminate inwardly of the pair of side walls to provide the passageway.

[0064] Embodiment 5 2. The cell culture vessel of embodiment 1, further comprising a lid sized and configured to cover the observation surface of the vessel body.

[0065] Embodiment 6 2. The cell culture vessel of embodiment 1, further comprising a protective carrier comprising a plate sized and configured to cover the cell culture surface of the vessel body.

[0066] Embodiment 7 7. The cell culture vessel of embodiment 6, wherein the protective carrier comprises a lip extending around the periphery of the plate sized and configured to extend along the first end wall, the end sidewall, and the pair of sidewalls.

[0067] Embodiment 8 7. The cell culture vessel of embodiment 6, wherein the protective carrier comprises standoff ribs positioned between the plate and the vessel body and arranged and configured to provide a space therebetween.

[0068] Embodiment 9 2. The cell culture vessel of embodiment 1, wherein the diverter extends at least about 50% to about 90% or less of the width of the inclined inner surface between opposite ends of the diverter.

[0069] Embodiment 10 2. The cell culture vessel of embodiment 1, wherein the vessel body further comprises a neck extending outward from the first end wall in a filling direction to an opening, the neck including a sloped filling surface having an outer edge at the opening that is angularly offset from an inner edge of the sloped filling surface at the first end wall.

[0070] Embodiment 11 11. The cell culture vessel of embodiment 10, wherein the inclined filling surface and the inclined inner surface intersect.

[0071] Embodiment 12 1. A method of forming a cell culture vessel, comprising: The container body is a first end wall; a second end wall opposite the first end wall; a pair of side walls extending between the first end wall and the second end wall, the first end wall, the second end wall and the pair of side walls defining an outer periphery of a cell culture chamber; an inner filler wall extending inwardly into the cell culture chamber from the first end wall and the pair of side walls, the inner filler wall having a beveled inner surface, the beveled inner surface having an outer edge at the first end wall that is angularly offset from an inner terminal edge of the beveled inner surface; a diverter extending along the inclined inner surface at least partially transverse to the filling direction between the pair of side walls, the diverter providing a passageway between the diverter and the pair of side walls through which liquid flows during a filling operation; forming the container body as a single monolithic piece, so as to comprise: forming a protective carrier separately from the vessel body, the protective carrier including a plate sized and configured to cover the cell culture surface of the vessel body; A method comprising:

[0072] Embodiment 13 13. The method of claim 12, wherein the entire length of the diverter is continuous with the inclined inner surface of the inner fill wall.

[0073] Embodiment 14 13. The method of claim 12, wherein the diverter has opposing ends that terminate inwardly of the pair of side walls to provide the passageway.

[0074] Embodiment 15 13. The method of embodiment 12, further comprising forming a lid configured to cover the viewing surface of the container body.

[0075] Embodiment 16 13. The method of claim 12, wherein the protective carrier comprises a lip extending around the periphery of the plate sized and configured to extend along the first end wall, the end side wall, and the pair of side walls.

[0076] Embodiment 17 13. The method of embodiment 12, wherein the protective carrier comprises standoff ribs positioned between the plate and the container body and arranged and configured to provide a space therebetween.

[0077] Embodiment 18 13. The method of claim 12, wherein the diverter extends at least about 50% to no more than about 90% of the width of the angled inner surface between opposite ends of the diverter.

[0078] Embodiment 19 13. The method of claim 12, wherein the container body further comprises a neck extending outward from the first end wall in a filling direction to an opening, the neck including a sloped filling surface having an outer edge at the opening that is angularly offset from an inner edge of the sloped filling surface at the first end wall.

[0079] Embodiment 20 20. The method of claim 19, wherein the inclined filling surface and the inclined inner surface intersect.

Claims

1. A cell culture vessel, comprising: A container body is provided, the container body comprising: a first end wall; a second end wall opposite the first end wall; a pair of side walls extending between the first end wall and the second end wall, the first end wall, the second end wall and the pair of side walls defining an outer periphery of a cell culture chamber; an inner filler wall extending inwardly into the cell culture chamber from the first end wall and the pair of side walls, the inner filler wall having a beveled inner surface, the beveled inner surface having an outer edge at the first end wall that is angularly offset from an inner terminal edge of the beveled inner surface; a diverter extending along the inclined inner surface at least partially transverse to the filling direction between the pair of side walls, the diverter providing a passageway between the diverter and the pair of side walls through which liquid flows during a filling operation; A cell culture vessel comprising:

2. The cell culture vessel of claim 1 , wherein the entire length of the diverter is continuous with the inclined inner surface of the inner filling wall.

3. The cell culture vessel of claim 2 , wherein the diverter is formed as an integral, monolithic part of the inner fill wall.

4. The cell culture vessel of claim 1 , wherein the diverter has opposing ends that terminate inwardly of the pair of side walls to provide the passageway.

5. The cell culture vessel of claim 1 , further comprising a lid sized and configured to cover the observation surface of the vessel body.

6. The cell culture vessel of claim 1 , further comprising a protective carrier comprising a plate sized and configured to cover the cell culture surface of the vessel body.

7. 7. The cell culture vessel of claim 6, wherein the protective carrier comprises a lip extending around the periphery of the plate sized and configured to extend along the first end wall, the end sidewall, and the pair of sidewalls.

8. The cell culture vessel of claim 6 , wherein the protective carrier comprises standoff ribs positioned between the plate and the vessel body and arranged and configured to provide a space therebetween.

9. 10. The cell culture vessel of claim 1, wherein the diverter extends from at least about 50% to no more than about 90% of the width of the inclined inner surface between opposite ends of the diverter.

10. 2. The cell culture vessel of claim 1, wherein the vessel body further comprises a neck extending outward from the first end wall in a filling direction to an opening, the neck including a sloped filling surface having an outer edge at the opening that is angularly offset from an inner edge of the sloped filling surface at the first end wall.

11. The cell culture vessel according to claim 10 , wherein the inclined filling surface and the inclined inner surface intersect.

12. 1. A method of forming a cell culture vessel, comprising: The container body is a first end wall; a second end wall opposite the first end wall; a pair of side walls extending between the first end wall and the second end wall, the first end wall, the second end wall and the pair of side walls defining an outer periphery of a cell culture chamber; an inner filler wall extending inwardly into the cell culture chamber from the first end wall and the pair of side walls, the inner filler wall having a beveled inner surface, the beveled inner surface having an outer edge at the first end wall that is angularly offset from an inner terminal edge of the beveled inner surface; a diverter extending along the entire length of the inclined inner surface of the inner filling wall at least partially transverse to the filling direction between the pair of side walls, the diverter providing a passageway between the diverter and the pair of side walls through which liquid flows during a filling operation; forming the container body as a single monolithic piece, so as to comprise: forming a protective carrier separately from the vessel body, the protective carrier including a plate sized and configured to cover the cell culture surface of the vessel body; A method comprising:

13. 13. The method of claim 12, wherein the protective carrier comprises a lip extending around the periphery of the plate sized and configured to extend along the first end wall, the end sidewall, and the pair of sidewalls, and further comprises standoff ribs positioned between the plate and the container body and arranged and configured to provide a space therebetween.

14. The method of claim 12, wherein the diverter extends at least about 50% to no more than about 90% of the width of the angled inner surface between opposite ends of the diverter.

15. 13. The method of claim 12, wherein the container body further comprises a neck extending outward from the first end wall in a filling direction to an opening, the neck including a sloped filling surface having an outer edge at the opening that is angularly offset from an inner edge of the sloped filling surface at the first end wall, the sloped filling surface intersecting the sloped inner surface.

Images