Becton Dickinson and Company Limited

Harry E Gray

Edward F Mussi

A roller bottle for trans-membrane co-culture of cells includes an exterior receptacle with a longitudinal axis having a first chamber surrounded by a sidewall. The exterior receptacle has a first neck at one end having an opening therethrough providing fluid access to said first chamber. The roller bottle further includes an interior container with a longitudinal axis and a second chamber, the interior container being located coaxially within the exterior receptacle. The interior container has a second neck at one end providing fluid access to the second chamber. The exterior receptacle is formed from a material that is substantially impermeable to gas and liquid and is sealed in a substantially fluid tight fashion forming the first chamber that has fluid access by the first neck. At least a portion of the interior container is formed from a microporous material. The roller bottle of the invention enables culturing one cell type in the presence of another cell type for the study of interactions between the cells while maintaining a physical separation between the cell types providing a scale up in cell population size from well type inserts. The microporous membrane allows free interchange of media soluble cellular products between the cell types, but maintains physical separation.

Roller bottle for trans-membrane co-culture of cells and method for its

A removable cell culture insert support device is provided which allows the initial development of cell cultures on the bottom membrane surface of an inverted cell culture insert. Following successful cell culture development, the support device of the invention is discarded and the conventional insert is reoriented and then suspended in a conventional multi-well plate with the initially developed cells on the bottom surface of the insert membrane. The upper surface of the membrane is then available for developing a second cell type for transmembrane interaction as required. The removable support includes a cup-shaped chamber with a surrounding skirt to accommodate the cell insert of interest, with the removable support having appropriate cooperating attaching surfaces. A double-sided adhesive may be used to temporarily join the two parts together.

Transmembrane cell culture device

A self-contained ready-to-use trans-membrane co-culture system of the present invention includes an insert having a tube with a longitudinal axis, a sidewall substantially parallel to the axis connecting a first open end and a second end. The second end has a microporous membrane bonded to it substantially perpendicular to the axis forming a well within the insert. The insert has a flange extending outwardly from the sidewall at the first end for suspending the insert as described hereinabove with a removable plug placed in the well of the insert to substantially prevent fluid flow through the microporous membrane. The insert with the plug is contained in a housing having a first end, a second end and a passageway therethrough. The housing has a longitudinal axis, a first diameter at the first end, a second diameter at the second end and an intermediate diameter between the first and second diameter. When the second end of the insert is coaxially placed in the first end of the housing, the second end of the insert has an interference fit with the intermediate diameter of the housing forming a chamber above the microporous membrane. The first and second ends of the housing are reclosably sealed with gas permeable lids that substantially prevent the passage of microorganisms into the sterililzed system housing as long as the lids are unopened. The present invention includes a method for trans-membrane co-culture of cells using the insert and the system of the present invention.

Device for trans-membrane co-culture of cells and a method for using same

A vessel and closure assembly for culturing mammalian cells wherein a gas permeable membrane is provided in the closure to allow rapid and uniform equilibration of gases between the atmosphere of the vessel and the atmosphere of an incubator. Removable thin impermeable film labels are provided for allowing passage of gases through the gas permeable membrane for uniform equilibration between the atmosphere in the closure and the atmosphere of the incubator as well as for occluding passage of gases through the gas permeable membrane when the vessel is removed from the controlled atmosphere of the incubator.

Vessel and closure assembly

A roller bottle is provided with provision for increased internal surface area for cell growth by the use of a foamed, textured or woven liner or sleeve inserted into the bottle prior to use. The sleeve may be used in a single use roller bottle or a multiple use roller bottle. With the latter, the sleeve is discarded, the bottle sterilized, and a new sleeve or liner is inserted. The interstices of the foamed, textured or woven surface has the effect of increasing the actual surface area available for culturing cells in each single roller bottle, thus making the roller bottle of the invention cost effective.

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Bloomingdale, NJ, United States of America

Harry E Gray