WO2017066821A1

WO2017066821A1 - Apparatus and products for culturing of cells or tissues

Info

Publication number: WO2017066821A1
Application number: PCT/AU2016/000359
Authority: WO
Inventors: John Greenwood
Original assignee: Skin Tissue Engineering Pty Ltd
Priority date: 2015-10-19
Filing date: 2016-10-19
Publication date: 2017-04-27

Abstract

An incubator for culturing cells or tissue, the assembly that includes a culture vessel holder for holding a number of culture vessels for culturing cells or tissue, the culture vessels comprising a fluid inlet permitting fluid to enter the culture vessel and a fluid outlet permitting fluid to be removed from the culture vessel and a waste container for fluid connection to the fluid outlet of one or more of the culture vessels to receive fluid removed from one or more of the culture vessels into the waste container and the waste container is held within the culture vessel holder.

Description

APPARATUS AND PRODUCTS FOR CULTURING OF CELLS OR TISSUES FIELD

[001 ] The present disclosure relates to products for culturing cells or tissues. BACKGROUND

[002] The growth of cells and tissue in vitro is widely undertaken. Typically, cells and tissue are grown in an incubator under suitable temperature, atmospheric and culture conditions.

[003] Under some circumstances there is a need to grow cells or tissue in larger quantities, particularly for medical applications.

[004] For example, the treatment of burns is an area which would benefit from the ability to grow large amounts of autologous cultured skin. Burns patients having an extensive part of their body affected cannot rely on the use of skin grafts and as such the production of large amounts of cultured autologous skin provides one solution to treating the burns.

[005] The large scale growth of cells and tissue provides a number of challenges, including the handling of the cultures and the possibility of contamination arising. These issues are in part a result of the difficulties associated with management of fluids during culturing.

[006] This is particularly the case for autologous cultured skin, as only a small sample of donor skin is generally used to seed and then expand the cells. As such, there is often a need to use a number of culture vessels to grow and expand the cells to a sufficient level to meet the coverage and timing requirements for a burns patient. Any loss of the amount of cultured skin produced due to handling difficulties and/or contamination can have profound effects on the patient.

[007] As such, there is a need for improved methods of culturing cells or tissue. [008]

SUMMARY

[009] Certain embodiments of the present disclosure provide an assembly for culturing cells or tissue, the assembly comprising:

a culture vessel holder for holding a plurality of culture vessels for culturing cells or tissue, the culture vessels comprising a fluid inlet permitting fluid to enter the culture vessel and a fluid outlet permitting fluid to be removed from the culture vessel; and a waste container for fluid connection to the fluid outlet of one or more of the culture vessels to receive fluid removed from one or more of the culture vessels into the waste container;

wherein the waste container is held within the culture vessel holder.

[0010] Certain embodiments of the present disclosure provide an assembly for culturing cells or tissue, the assembly comprising:

a culture vessel holder for holding a plurality of culture vessels for culturing cells or tissue,

one or more culture vessels held in the culture vessel holder, the culture vessels comprising a fluid inlet permitting fluid to enter the culture vessel and a fluid outlet permitting fluid to be removed from the culture vessel; and

a waste container for fluid connection to the fluid outlet of one or more of the culture vessels to receive fluid removed from one or more of the culture vessels into the waste container;

wherein the waste container is held within the culture vessel holder.

[001 1] Certain embodiments of the present disclosure provide an incubator for culturing cells or tissue, the incubator comprising an assembly comprising:

a culture vessel holder for holding a plurality of culture vessels for culturing cells or tissue, the culture vessels comprising a fluid inlet permitting a fluid to enter the culture vessel and a fluid outlet pennitting the fluid to be removed from the culture vessel; and

a waste container for fluid connection to the fluid outlet of one or more of the culture vessels to receive the fluid removed from one or more of the culture vessels into the waste container;

wherein the waste container is held within the culture vessel holder.

[0012] Certain embodiments of the present disclosure provide an incubator for culturing cells or tissue, the incubator comprising an assembly comprising:

one or more culture vessels held in the culture vessel holder, the culture vessels comprising a fluid inlet permitting a fluid to enter the culture vessel and a fluid outlet permitting the fluid to be removed from the culture vessel; and

wherein the waste container is held within the culture vessel holder.

[0013] Certain embodiments of the present disclosure provide a culture vessel comprising a fluid inlet permitting fluid to enter the culture vessel and a fluid outlet permitting fluid to exit the culture vessel.

[0014] Certain embodiments of the present disclosure provide a culture vessel comprising a removable platform for growing cells or tissue on the platform or for growing cells or tissue on a scaffold supported by the removable platform.

[0015] Other embodiments are disclosed herein.

BRIEF DESCRIPTION OF THE FIGURES

[0016] Certain embodiments are illustrated by the following figures. It is to be understood that the following description is for the purpose of describing particular embodiments only and is not intended to be limiting with respect to the description.

[0017] Figure 1 shows an external elevated view of a culture vessel according to one embodiment. [0018] Figure 2 shows an external elevated view of a culture plate according to one embodiment, in which the slidable flap covering the fluid inlet has been removed.

[0019] Figure 3 shows an external elevated view of a culture plate according to one embodiment with the lid removed.

[0020] Figure 4 shows an exploded view of the separate components of a culture plate according to one embodiment.

[0021] Figure 5 shows an elevated perspective view of an assembly for culturing cells or tissue according to one embodiment.

[0022] Figure 6 shows an elevated perspective view of an empty culture plate holder with waste management components shown according to one embodiment.

[0023] Figure 7 shows an elevated perspective view of an assembly with the manifold boot and waste hoses removed and in which the culture plates have been tilted to drain fluid from the plates.

[0024] Figure 8 shows an elevated perspective view of a fluid delivery means according to one embodiment.

[0025] Figure 9 shows an elevated perspective view of a tower holding two assemblies for use in an incubator.

[0026] Figure 10 shows an elevated perspective view of two towers and a medium reservoir assembly for supplying culture medium to the culture plates according to one embodiment.

[0027] Figure 1 1 shows an elevated perspective view of a bioreactor assembly according to one embodiment. DETAILED DESCRIPTION

[0028] The present disclosure relates to an assembly for cultunng cells or tissue. The present disclosure also relates to an incubator for culturing cells or tissue using the assembly, culture vessels for culturing cells or tissue, and methods for culturing cells or tissue and methods for managing or controlling fluid exchange.

[0029] The present disclosure arises from investigations into the need for providing sufficient amounts of skin for the treatment of burns. For the treatment of burns patients with extensive burns, the ability to utilise skin grafts is limited.

[0030] To address this issue, a means for culturing autologous skin cells and composites from a small amount of donor skin has been developed, which provides a greatly increased amount of composite skin that can be used.

[0031 ] However, it has also been recognised that the need to greatly expand skin cells and culture large pieces of composite skin presents difficulties with respect to fluid management and/or contamination of the growing cells.

[0032] Certain embodiments of the present disclosure provide an assembly for culturing cells or tissue, which provides a modular solution to the growth of cells or tissue and provides benefits to management of fluids for culturing cells or tissue, and/or reduces contamination.

[0033] Certain embodiments of the present disclosure provide an assembly for culturing cells or tissue, the assembly comprising:

wherein the waste container is held within the culture vessel holder.

[0034] The term "fluid" refers to a substantially liquid medium, and in some examples is a culture medium, a wash medium, or a solution. Other types of fluids are contemplated.

[0035] In certain embodiments, the culture vessels comprise a culture plate. Other types of culture vessels are contemplated, such as a culture dish or a culture flask.

[0036] In certain embodiments, the assembly comprises a manifold for fluid connection of the fluid outlet of one or more of the culture vessels to the waste container. In certain embodiments, the manifold is connected to a fluid outlet on each of the culture vessels.

[0037] In certain embodiments, the manifold is removably connected to a fluid outlet.

[0038] In certain embodiments, the manifold is removably connected to the waste container and/or a hose connected thereto.

[0039] In certain embodiments, the manifold is connected to a single hose that is connected to the waste container.

[0040] In certain embodiments, the assembly comprises one or more hoses.

[0041] In certain embodiments, the assembly comprises one or more hoses for fluid connection of a culture vessel to the waste container.

[0042] In certain embodiments, the culture vessel holder holds 2 or more culture vessels. In certain embodiments, the culture vessel holder holds 5 or more culture vessels. Other numbers of culture vessels are contemplated.

[0043] In certain embodiments, the waste container comprises a bag, such as bladder bag. In certain embodiments, the waste container comprises a rigid container.

[0044] In certain embodiments, the waste container is removable from the culture vessel holder. In certain embodiments, the waste container is a replaceable component.

[0045] In certain embodiments, the waste container is located below the level of the culture vessel. This arrangement permits gravity feed of fluid removed from a culture vessel into the waste container. [0046] In certain embodiments, the waste container is located at the base of the culture vessel holder. In certain embodiments, the culture vessel holder comprises a tray at the base of the holder and the waste container sits in the tray.

[0047] In certain embodiments, the fluid inlet and the fluid outlet are located substantially opposed to one another. This provides a benefit to cross-flow refreshment of fluid in the culture vessel.

[0048] In certain embodiments, the culture vessel holder comprises a means for supporting the culture vessels. In certain embodiments, the means for supporting culture vessels comprises one or more vertical panels with ledges.

[0049] In certain embodiments, the holder comprises one or more guides to assist with guiding the culture vessels into the culture vessel holder.

[0050] In certain embodiments, the vertical panels comprise laterally displaced front tabs to assist with guiding the culture vessels into the culture vessel holder and/or prevent the culture vessels from being withdrawn.

[0051] In certain embodiments, the culture holder is configured to permit tilting of the culture vessels in the holder so as to remove fluid from the culture vessels via the fluid outlet upon tilting of the culture vessels.

[0052] In certain embodiments, the culture vessel holder comprises a means for supporting the culture vessels and the means for supporting is pivotally connected to the holder to permit tilting of the culture vessels. In certain embodiments, the means for supporting culture vessels comprises one or more vertical panels and the panels are pivotally connected to the culture vessel holder.

[0053] In certain embodiments, the tilting of the culture vessels is biased towards the horizontal position. This assists with holding the culture vessels in the horizontal position and with return of the culture vessels to the horizontal position once tilting has occurred.

[0054] In certain embodiments, the assembly comprises a resilient component and/or a spring to bias the tilting of the culture vessels towards the horizontal position. [0055] Certain embodiments of the present disclosure provide an assembly for culturing cells or tissue, the assembly comprising:

wherein the waste container is held within the culture vessel holder.

[0056] Certain embodiments of the present disclosure provide use of an assembly as described herein to culture cells or tissue.

[0057] Certain embodiments of the present disclosure provide a method of growing cells or tissue by using an assembly as described herein.

[0058] Certain embodiments of the present disclosure provide a method of controlling fluid exchange in a culture vessel by using an assembly as described herein.

[0059] Certain embodiments of the present disclosure provide a method of controlling fluid exchange in a culture vessel, the method including using an assembly as described herein to control fluid exchange in the culture vessel.

[0060] Certain embodiments of the present disclosure provide an incubator comprising an assembly as described herein.

[0061 ] Certain embodiments of the present disclosure provide an incubator for culturing cells or tissue, the incubator comprising an assembly comprising:

a culture vessel holder for holding a plurality of culture vessels for culturing cells or tissue, the culture vessels comprising a fluid inlet permitting a fluid to enter the culture vessel and a fluid outlet permitting the fluid to be removed from the culture vessel; and

wherein the waste container is held within the culture vessel holder.

[0062] Culture vessel holders are as described herein. In certain embodiments the culture vessel holder holds one or more culture vessels. In certain embodiments, the culture vessels comprise culture plates or culture flasks.

[0063] In certain embodiments, the culture vessel holder is configured to hold a culture vessel having a top and/or a bottom surface area of greater than 400 cm².

[0064] In certain embodiments, the culture vessel holder holds 5 or more culture vessels. Other numbers of culture vessels are contemplated.

[0065] Assemblies are as described herein. In certain embodiments, the incubator comprises a plurality of assemblies, for example 2 or more assemblies, 3 or more assemblies or 4 or more assemblies.

[0066] In certain embodiments, an assembly comprises a manifold for fluid connection of the fluid outlet of one or more of the culture vessels to the waste container. In certain embodiments, the manifold is removably connected to the fluid outlet. In certain embodiments, the manifold is removably connected to the waste container and/or a hose connected thereto.

[0067] Waste containers are as described herein. In certain embodiments, the waste container is removable from the culture vessel holder.

[0068] Fluid inlets and fluid outlets of the culture vessels are as described herein. In certain embodiments, the fluid inlet and the fluid outlet are located substantially opposed to one another. In certain embodiments, the fluid inlet and the fluid outlet are located adjacent to one another. [0069] In certain embodiments, the culture vessel holder is configured to permit tilting of the culture vessels in the holder so as to remove fluid from the culture vessels via the fluid outlet upon tilting of the culture vessels.

[0070] In certain embodiments, the culture vessel holder comprises a means for supporting the culture vessels and the means for supporting is pivotally connected to the holder to permit tilting of the culture vessels.

[0071] In certain embodiments the tilting of the culture vessels is biased towards the horizontal position. In certain embodiments, the assembly comprises a resilient component and/or a spring to bias the tilting of the culture vessels towards the horizontal position. In certain embodiments, the culture vessel holder comprises a resilient component and/or a spring to bias the tilting of the culture vessels towards the horizontal position.

[0072] In certain embodiments, a mechanical means is used to tilt the culture vessels.

[0073] In certain embodiments, the incubator, or an assembly containing an incubator, comprises a mechanical means to tilt the culture vessel.

[0074] In certain embodiments, the incubator comprises a fluid input means to deliver fluid to the culture vessels through the fluid inlet. In certain embodiments, the fluid input means comprises a spigot. In certain embodiments the fluid input means comprises a tube. In certain embodiments, the fluid input means is used to deliver fluid to a plurality of culture vessels. In certain embodiments, the fluid input means delivers fluid to individual culture vessels.

[0075] In certain embodiments, the fluid input means is mechanically controlled. In certain embodiments, the fluid inlet comprises a means for sealing the inlet. In certain embodiments, the fluid input means engages with the means for sealing the inlet to deliver fluid to the culture vessel.

[0076] In certain embodiments, the means for sealing the fluid inlet comprises a moveable flap. In certain embodiments, the fluid input means moves the flap to an open position to deliver fluid and the flap returns to a sealed position upon withdrawal of the fluid input means. [0077] In certain embodiments, the incubator comprises one or more fluid reservoirs. In certain embodiments, the one or more fluid reservoirs comprise a culture medium reservoir and/or wash medium.

[0078] In certain embodiments, the incubator comprises means to mechanically control delivery and/or removal of the fluid to the culture vessels. In certain embodiments, the incubator comprises computer controlled means to automatically deliver and/or remove fluid. In certain embodiments, the incubator (or an ancillary device) provides computer controlled management of fluid exchange for the culture vessels.

[0079] Certain embodiments of the present disclosure provide a culture vessel.

[0080] In certain embodiments, the culture vessel is used in an assembly as described herein.

[0081 ] In certain embodiments, the culture vessel is used in an incubator or a bioreactor as described herein. The term "bioreactor" as used herein refers to a system for culturing cells and tissue, and typically includes an incubator for providing suitable temperature and/or atmospheric conditions and one or more other components, such as components involved in fluid management.

[0082] Certain embodiments of the present disclosure provide a culture vessel comprising a fluid inlet permitting fluid to enter the culture vessel and a fluid outlet permitting fluid to exit the culture vessel.

[0083] In certain embodiments, the fluid inlet comprises a means for sealing the fluid inlet. In certain embodiments, the means for sealing the fluid inlet comprises a moveable flap. In certain embodiments, wherein movement of the moveable flap from a sealed position to an open position permits delivery of fluid through the fluid inlet. In certain embodiments, the moveable flap comprises a resilient means to return the moveable flap from the open position to the sealed position. In certain embodiments, the moveable flap is a slidable or a rotatable flap.

[0084] In certain embodiments, the culture vessel comprises a culture plate. In certain embodiments, the culture vessel comprises a culture flask. Other types of culture vessels are contemplated. [0085] In certain embodiments, the culture vessel comprises a culture plate with a base and a removable lid.

[0086] In certain embodiments, the lid of the culture plate comprises a fluid inlet. In certain embodiments, the base of the culture plate comprises a fl uid outlet.

[0087] In certain embodiments, the culture vessel comprises a means to reduce fluid velocity of fluid entering the culture vessel through the fluid inlet. In certain embodiments, the means to reduce fluid velocity comprises one or more surfaces. In certain embodiments, the means to reduce fluid velocity comprises one or more surfaces above the level of cell or tissue culturing. In certain embodiments, the means to reduce fluid velocity comprises one or more baffles. In certain embodiments, the means to reduce fluid velocity comprise one or more barriers.

[0088] In certain embodiments, the culture vessel comprises one or more internal barriers to hold a desired level of fluid in the culture vessel. In certain embodiments, the one or more internal barriers prevent fluid from flowing out the fluid outlet. In certain embodiments, one of the internal barriers comprises one or more recesses below the level of the top of the other internal barriers.

[0089] In certain embodiments, the culture vessel comprises a removable platfonn for growing cells or tissue on the platform or for growing cells or tissue on a scaffold supported by the removable platform. In this regard, it will be appreciated that the culture vessels as described herein are suitable for growing cells or tissue. In an embodiment where the culture vessels further comprises a removable platform, cells or tissue may be grown on the removable platfonn directly, or grown on a scaffold which sits on the platfonn.

[0090] In certain embodiments, the removable lid comprises one or more projections to hold the removable platfonn under fluid (eg culture medium) in use.

[0091] Certain embodiments of the present disclosure provide a culture vessel comprising a removable platfonn for growing cells or tissue on the platform or for growing cells or tissue on a scaffold supported by the removable platform.

[0092] In certain embodiments, the culture vessel comprises a culture plate with a base and a removable lid.

[0093] In certain embodiments, the removable lid comprises one or more projections to hold the removable platfonn under the fluid (eg a culture medium) in use.

[0094] In certain embodiments, the culture vessel comprises a fluid inlet permitting fluid to enter the culture vessel and a fluid outlet permitting fluid to exit the culture vessel.

[0095] In certain embodiments, the culture vessel comprises a culture plate with a base and a removable lid and the lid comprises the fluid inlet,

[0096] In certain embodiments, the culture vessel comprises a culture plate with a base and a removable lid and the base comprises the fluid outlet.

[0097] In certain embodiments, wherein the culture vessel comprises a means to reduce fluid velocity entering the culture vessel through the fluid inlet.

[0098] In certain embodiments, the means to reduce fluid velocity comprises one or more surfaces above the level of cell or tissue culturing and/or one or more baffles. In certain embodiments, the means to reduce fluid velocity comprise one or more barriers.

[0099] In certain embodiments, the culture vessel comprises one or more internal barriers to hold a desired level of fluid in the culture vessel. In certain embodiments, the one or more barriers prevent fluid from flowing out a fluid outlet.

[00100] In certain embodiments, a barrier comprises one or more recesses below the level of the top of the other internal barriers.

[00101] In certain embodiments, the fluid inlet comprises a means for sealing the fluid inlet. In certain embodiments, the culture vessel comprises a moveable flap to seal the fluid inlet. In certain embodiments, movement of the moveable flap from a closed position to an open position permits delivery of fluid through the fluid inlet.

[00102] In certain embodiments, the moveable flap comprises a resilient means to return the moveable flap from the open position to a closed position. In certain embodiments, the moveable flap is a slidable flap.

[00103] Certain embodiments of the present disclosure provide a method of culturmg cells or tissue, the method comprising incubating the cells or tissue in an incubator as described herein.

[00104] Certain embodiments of the present disclosure provide a method of culturing cells or tissue, the method comprising culturing the cells or tissue in a culture vessel as described herein.

[00105] Certain embodiments of the present disclosure provide a bioreactor comprising an assembly for culturing cells or tissue as described herein.

[00106] In certain embodiments, the bioreactor comprises an incubator and/or a fluid delivery means for delivering fluid to the assembly. Incubators and fluid delivery means are as described herein.

[00107] In certain embodiments, the bioreactor comprises a component as described herein.

[00108] Certain embodiments of the present disclosure provide a bioreactor for culturing cells or tissue, the bioreactor comprising an assembly for culturing cells or tissue, an incubator and a fluid delivery means for delivering fluid to the assembly.

[00109] Certain embodiments of the present disclosure provide a method of culturing cells or tissue, the method comprising culturing the cells or tissue in a bioreactor as described herein.

[00110] The present disclosure is further described by the following description of specific embodiments. It is to be understood that the following description is for the purpose of describing particular embodiments only and is not intended to be limiting with respect to the above description.

DETAILED DESCRIPTION OF SPECIFIC EMBODIMENTS

[001 1 1 ] Figure 1 shows an external elevated view of a culture vessel according to one embodiment. [001 12] The culture vessel may be used to culture cells or tissue, for example to culture skin cells in an incubator or a bioreactor using suitable media and culture conditions, according to one embodiment.

[001 13] In the embodiment shown, the culture vessel is a culture plate 1 10. The culture plate 1 10 comprises a removable lid 1 1 1 and a base 1 13. The lid 1 1 1 and base 1 13 may be made from a suitable material, for example a plastic such as polycarbonate plastic,

[001 14] The culture plate 1 10 comprises one or more fluid inlets (1 16 and 1 17), which permit fluid to enter the culture plate 1 10, and one or more fluid outlets (1 18 and 119) which permit fluid to exit or be removed from the culture plate 1 10.

[00115] In the embodiment shown, the culture plate comprises two fluid inlets 116 and 1 17 (not visible). Fluid inlet 116 is located in the base 1 13 and is integral with the base 1 13. Fluid inlet 1 16 permits delivery of fluid to the plate. Fluid inlet 1 16 can be connected to a hose, tube or pipe for entry of fluid into the plate 1 10. For example, the fluid inlet 1 16 can be mated with a tube connected to a peristaltic pump tube to deliver fluid to the culture plate 110.

[001 16] Fluid inlet 1 17, in the form of an inlet port, is located in the lid 1 1 1 and is covered by a sprung moveable flap 120. The moveable flap acts to cover and seal the port 1 17 when the flap 120 is in the closed/sealed position. Movement of the moveable flap 120 to an open position pennits delivery of fluid through the fluid inlet 1 17. The moveable flap may be made from a suitable materials, such as a plastic,

[001 17] The moveable flap 120 is slidable and covers the fluid inlet in the closed position shown, thereby sealing the fluid inlet 1 17, and pivots laterally about 121 to expose the fluid inlet 1 17 underneath and thereby permit deli very of fluid into the plate 1 10.

[001 18] The slidable flap 120 is typically made from a resilient plastic and the flap 120 comprises a resilient an 122. The lid 1 1 1 comprises an elongate trough 123 which receives the end 124 of the resilient arm 122 and pennits the end 124 to slide along the trough 123 when the ann is compressed. The lid 1 10 also comprises a barrier 125 against which the end 124 abuts. The slidable flap 120 may be pushed laterally towards the centre of the plate 1 10 so as to expose the fluid inlet 1 17. As a consequence, the resilient ann 122 is compressed and once the lateral force that has moved the slidable flap 120 to the open position is removed, the resilient ann 122 forces the slidable flap 120 back to the closed or sealed position shown and thereby seals the fluid inlet 1 17.

[001 19] The lid 1 1 1 is secured to the base 1 13 by one or more webs that slide below the top flange of base 1 13 and by two latches on the lid (not visible) opposed to fluid inlets 1 16 and 117. The lid also comprises tabs 1 14 and 1 15 to hold the lid clear of a bench when it is put down, thus reducing the chances of contamination.

[00120] The culture plate comprises a base 113 which comprises one or more fluid outlets. In the embodiment shown, the culture plate 1 10 comprises two fluid outlets 1 18 and 1 19. One fluid outlet 1 19 is located opposite the fluid inlets 1 16 and 1 17, and the other fluid outlet 1 18 is located on the same side of the plate 1 10 as fluid inlets 1 16 and 1 17. The outlets 1 18 and 1 19 permit removal of waste fluid from the culture plate 1 10, which in the embodiment shown is achieved by tilting of the plate towards either of the fluid outlets 1 18 or 1 19. The fluid outlets 1 18 and 1 19 can be connected to a hose, tube or pipe for removal of fluid from the plate 1 10. For example, the fluid outlet 1 18 and 1 19 can be mated with a tube or with a custom manifold.

[00121 ] In one embodiment, the culture plate 1 10 may be used for culturing autologous skin or autologous composite skin, used for example to treat burns or wounds and/or in reconstructive or plastic surgery. For these purposes the culture plates would generally have a size of at least 20 cm x 20 cm, and typically of a size of 25 cm x 25 cm.

[00122] In the embodiment shown, the culture plate 1 10 also comprises a removable platform 136 for growing cells or tissue on the platform 136 or for growing cells or tissue on a scaffold supported by the removable platform 136. The use of removable platfonn assists with removal of cells or tissue from the plate 1 10, for example where skin cells are grown on a scaffold to form a cultured composite skin and the culture composite skin alone needs to be conveyed to a surgeon using the cultured composite skin to treat a wound or burn. The removable platform may be made of suitable material, such as the same material from which the plate is made (eg a polycarbonate), or another material. [00123] Figure 2 shows an external elevated view of a culture plate 210 for use in an incubator or bioreactor according to one embodiment, in which the slidable flap covering the fluid inlet 217 shown in Figure 1 has been removed.

[00124] The lid 21 1 comprises an aperture 226, adjacent to the fluid inlet 217. The aperture 226 receives a downward projection on the end of the slidable flap which allows the slidable flap to pivot about the aperture 226. When the slidable flap moves from the closed/sealed position to the open position, by pivoting about 226, the fluid inlet 217 is exposed and permits fluid to be delivered via inlet 217.

[00125] The lid 21 1 also comprises an elongate trough 223, which receives the end of the resilient arm on the slidable flap, and a barrier 225 against which the end of the resilient arm abuts.

[00126] Figure 3 shows an external elevated view of a culture plate 310 according to one embodiment with the lid removed.

[00127] The culture plate comprises a fluid inlet 316 which can be mated with a suitable hose, tube, pipe or connector.

[00128] The plate 310 also comprises two fluid outlets, 318 and 319. One fluid outlet 319 is located opposite the fluid inlet 316, and the other fluid outlet 318 is located on the same side of the plate 310 as fluid inlet 316. The fluid outlets 318 and 319 can be connected to a connector such as a suitable hose, tube, pipe or manifold for removal of the fluid from the plate 310. Fluid outlets 318 and 319 also have an O-ring 326 to assist with sealing of the outlets to a suitable connector and/or caps when not in use.

[00129] In the embodiment shown, the plate 310 comprises two internal barriers 327 and 328, which in conjunction with the side walls 329 of the plate 310 act to hold a desired level of fluid in the plate 310. Internal barrier 327 comprises an inner wall running across the base 313 of the plate 310, which acts to prevent fluid flowing out the fluid outlet 319. Internal barrier 328 comprises a wall with a sloping surface running across the base 313.

[00130] The height of the two internal barriers 327 and 328 is lower than the height of side walls 338, and accordingly the volume of fluid held in the plate 310 is dependent upon the height of the internal barriers 327 and 328. In the embodiment shown, the inner wall 327 comprises two or more recesses/spillways 330, which are below the level of the top of the other internal barriers and function to assist with releasing fluid over the wall as the fluid level rises and the meniscus eventually breaks over the recesses/spillways 330. This ensures a maximum level of fluid in the plate 310 which is below the top of the side walls 329 and also ensures that any excess fluid first breaks over the inner wall 327 first and the fluid can then exit from fluid outlet 319.

[00131 ] When plate 310 is holding a fluid, such as a culture medium, removal of the fluid from the plate 310 may be achieved by tilting the plate 310 towards fluid outlet 318 or tilting the plate 310 towards fluid outlet 319.

[00132] Tilting of the plate 310 towards fluid outlet 319 causes the fluid to flow over the internal wall 327. The fluid then will flow into the space/drain 331 between the internal wall 327 and the back wall 332 of the plate. The lowest point of fluid outlet 319 is coincident with the bottom of the plate 333, which results in fluid being removed from the plate 310 when titled.

[00133] Tilting of the plate 310 towards fluid outlet 318 causes the fluid to flow over the sloping surface of the internal wall 328 into the space/drain 334 between the inner wall 327 and the front wall 335. The lowest point of fluid outlet 318 is coincident with the bottom of the plate 333, which results in fluid being removed from the plate 310 when titled.

[00134] Waste elimination from the plate 310 can therefore be achieved in three ways: (1 ) tilting/tipping the plate downward at the end furthest from the fluid inlet 316. In the embodiment shown, tilting down 7 degrees results in fluid being eliminated. (2) tilting/tipping the plate downward at the same end as the fluid inlet 316. In the embodiment shown, tilting by down 7 degrees also results in fluid being eliminated. (3) constant replenishment of the fluid causing overflow of the fluid into one or both drains of drain 331 and 334.

[00135] The use of the fluid outlet 319 opposite the fluid inlet 316 or the fluid inlet in the lid facilitates cross-flow refreshment of fluid in the plate 310. [00136] In the embodiment shown, the plate 310 between internal walls 327 and 328 and the side walls 329 has a nominal fluid (eg culture medium) volume of approximately 500 millilitres. This volume is controlled by the height of the recesses/spillways 330 into the two drains 331 and 334. The nominal height of the overflow is 7.5mm above the bottom of the floor 333.

[00137] In the embodiment shown, the culture plate also comprises a removable platform 336 for growing cells or tissue on the platform 336 or for growing cells or tissue on a scaffold supported by the removable platform. The platform 336 comprises a cell or tissue growth area of 250 mm x 250 mm surrounded by a fluid/culture medium flow channel 337 in the bottom 333 of the plate 310, which is typically of a size of 1.5 mm x 1.5 mm, to ensure full immersion of a cells or tissue.

[00138] In the embodiment shown, fluid may enter the plate via fluid inlet 316 or the fluid inlet located in the lid (not shown). In the embodiment shown, the base 313 of the plate 310 also comprises a means 338 to reduce fluid velocity of fluid entering the culture plate through either of the fluid inlets. The means 338 also assists to disperse the flow of fluid entering the plate. In the embodiment shown, the means 338 comprises a plurality of baffles/barriers 339 which impede fluid flow and necessitate flow of fluid around the baffles/barriers 339 and the surface upon which the baffles/barriers are located, which acts to reduce fluid velocity as the fluid meets the surface. In addition, the means 338 is adjacent the sloping surface of the front inner wall 328, which assists with a more gradual introduction of fluid onto the growth area 336.

[00139] Refreshment of fluid, such a culture medium, in the plate 310 can be achieved either by way of tilting and draining the majority of spent fluid and then re-filling the culture medium through fluid inlet 316 or the fluid inlet in the lid, or by dilution of the fluid in the culture plate by periodic or constant refreshment of new fluid through the fluid inlets.

[00140] Figure 4 shows an exploded view of the separate components of a culture plate 410 according to one embodiment.

[00141 ] The culture plate 410 comprises a removable lid 411 and a base 413. The lid 41 1 comprises a fluid inlet 417. A movable flap 420 is used to seal the fluid inlet 417, and comprises a resilient arm 422. The moveable flap 420 comprises a downward extending projection 440 which engages with the aperture 426 on the lid to permit pivoting of the flap 420 in the lid 41 1.

[00142] The fluid inlet 416 has a cap 441. Fluid outlets 418 and 419 have caps 441 and O- rings 426.

[00143] In the embodiment shown, the culture plate 410 also comprises a removable growth platform 436 for culturing cells or tissue on the platform 436 if desired, or for supporting the growth of cells or tissue on a scaffold supported by the removable platform 436.

[00144] In one embodiment, cells are incorporated into a scaffold, such as a biodegradable polyurethane scaffold, and cultured on the platform 436 to support the scaffold. Removal of the scaffold with incorporated cells may then be achieved by removing the platform 436 from the culture plate 410. In this case, removal of the platform 436 is assisted by the presence of tab 442, which permits an instrument to be slid under the tab 442 and lift the platform 436.

[00145] The removable growth platform 436 also comprises side extending tabs 443 which ensure cells or tissue growing on the plate are fully surrounded by fluid (eg culture medium).

[00146] The lid 41 1 is also fitted with hold down tabs or projections 444 that contact tabs 443 to ensure that the growth platform is fully immersed in fluid, such as culture medium. Thus, the tabs or projections act to hold removable platform 436 under the fluid/culture medium in use.

[00147] The growth platform 436 also comprises a removal upwardly projecting element 446, such as a spacer, which engages to contact with a projection 446 on the lid 41 1 , to further ensure the submersion of the growth platform 468.

[00148] For example, when a 1 mm deep growth platform 436 supports a 1 mm thick scaffold of polyurethane foam, there is a maximum depth of 5.5mm of culture medium over the cells or tissue.

[00149] Referring to Figure 5, there is shown in one embodiment an elevated perspective view of an assembly 550 for culturing cells or tissue. [00150] The assembly 550 represents a modular unit for culturing cells or tissue and controlling fluid exchange in a culture vessel, such as a culture plate.

[00151 ] In one embodiment, an assembly may be used to culture autologous cells or tissue for introduction into a single patient. Typically, the assembly is used to culture cells or tissue in culture plates of a size of at least 400 cm² and usually about 25 cm x 25 cm in size.

[00152] The assembly 550 comprises a culture plate holder 551 which holds a plurality of culture plates 510 as described above. In the embodiment shown, the culture plate holder holds 5 plates, although the assembly can be readily adapted to holding fewer or more plates.

[00153] The culture plates comprise fluid inlet 516 and one not visible, which permit fluid to enter a culture plate 510. Fluid inlet 517 is sealed by a moveable flap 520.

[00154] The culture plates also comprise fluid outlets 518 and 519, which permit to fluid to be removed from the culture plate 510. In the embodiment shown, fluid inlets 516 and 517 are located opposed to fluid outlet 519.

[00155] In the embodiment shown, the assembly comprises a manifold 552 and a manifold boot 553. The manifold 552 is connected to each of the fluid outlets 519 to receive fluid from each of the culture plates 510. The manifold boot 553 is connected to a waste hose 554 which is connected via a waste container 555 (in the form of a waste bag) via a waste bag boot 556. The waste bag 555 is located in a tray 557 at the bottom and held within the culture plate holder 551.

[00156] In the embodiment shown, the manifold 552 is removably connected to the fluid outlets 519 and is removably connected to the waste container 555 via the waste hose 554. The manifold 552, manifold boot 553, waste hose 554, waste boot 556 and waste container 555 may all be separately replaceable components. It will be appreciated that while the use of a manifold is described in this embodiment, the fluid outlets 518 and 519 can alternatively be connected to tubes, hoses, pipes etc.

[00157] In the embodiment shown the assembly 550 is configured to permit tilting of the culture plates 510 in the holder 551 so as to remove the fluid from the culture plates via the fluid outlets 519 upon tilting of the culture plates 510. Removal of fluid from the culture plates 510 is achieved by tilting of the culture plates in the holder towards fluid outlets 519, each of which is connected to the manifold 552 and resulting in the waste fluid being stored in the waste container 555. The waste container 555 may be removed from the assembly 550 and either waste fluid removed or a new waste container may be installed.

[00158] In the embodiment shown, the waste container 555 is located below the level of the culture plates 510, being located at the base of the holder 551 in the tray 557. The waste container is removable from the holder 551 and can be a replaceable component if desired.

[00159] Figure 6 shows an elevated perspective view of an empty culture plate holder 1651 with waste management components shown (manifold 652, manifold boot 653, waste hose 654, waste boot 656 and waste container 655) according to one embodiment.

[00160] The culture plate holder 651 comprises a shoe 658, which is typically made from metal and produced as a single component having two sides 659 and a tray 657. The waste container 655 is held within the holder 658 in tray 657 at the bottom of the shoe 658.

[00161 ] In the embodiment shown, the culture plate holder 651 is configured to permit tilting of the culture plates (not shown) in the holder 651 so as to remove fluid from the culture plates via the fluid outlet, upon tilting of the culture vessels.

[00162] In the embodiment shown, the culture plate holder 651 comprises two upright or vertical panels 660 which are used for supporting the culture plates. The upright panels 660 are pivotally connected to the sides 659 of the culture plate holder 651 , to permit tilting of the culture plates in the holder 651. The panels 660 comprise a cylindrical projection 661 which fit into a U-shaped void at the top of the sides 659, thereby permitting the panels 660 to tilt.

[00163] The panels 660 are further connected by a cross brace 695, which contains a slot at each end to fit over the top of the sides 659 and thereby hold the cylindrical projection 661 in the sides 659. A connective plate 694, typically made of metal, is located at the base of the panels 660 and is a friction fit into slots at the base of the panels 660 to hold them at their correct spacing even when no culture vessels are held in the folder 651.

[00164] The upright panels 660 also each comprise a ledge 662 for supporting one side of a culture plate. [00165] The culture plate holder also comprises one or more guides 663 to assist with guiding the culture vessels into the culture vessel holder and which also act as flexible latches to clip the culture plates in place in the panels 660. The guides protrude forward so that an operator can use their finger tips to open the guides and slide the culture plate out.In the embodiments shown, the vertical panels 660 comprise front tabs 663, which are laterally displaced at their end and have a latch 664, to clip to a web on the base of the culture plate.

[00166] In the embodiment shown, the tilting mechanism is biased towards the horizontal position. This assists with holding the culture plates in the horizontal position and also assists with return of the culture plates to the horizontal position once tilting has occurred.

[00167] In the embodiment shown, the culture holder comprises two resilient components 665, in the form of resilient members, which are each connected to a side 659 and engage with the bottom of the panel 660 to urge the panel upward when tiled downwards. A spring 666 replaces and/or augments resilient member 665 to adjust the return force exerted on panels 660. As the vertical panel 660 is tilted downward, the spring compresses and provides further elastic tension.

[00168] Figure 7 shows elevated perspective view of an assembly 750 according to one embodiment, without showing the fluid connections to the waste container 755, in which the culture plates have been tilted to drain fluid from the plates 710.

[00169] In the embodiment, the vertical panels 760 have been tilted towards fluid outlet 719 at the reai" of the plates. The culture plate holder 751 remains in a fixed position, but the vertical panels 760 and plates 710 are now tilted relative to the holder 751.

[00170] Figure 8 shows a fluid delivery means 870 according to one embodiment. The fluid delivery means is connected to a fluid delivery tube 871 , connected to a fluid reservoir, such as a bag of culture medium. In the embodiment shown, the fluid delivery means comprises a spigot 872, which utilises a diaphragm 873 to control fluid entry to a culture vessel. Pumping of fluid into the spigot 872 raises the diaphragm 873 and thereby opens the spigot to allow the delivery of fluid.

[00171] The spigot 872 comprises a mounting means 874 which can be received by an adaptor mount 875, which is mechanically controlled so as to position and move the spigot to deliver fluid to a culture plate.

[00172] In the embodiment shown, the adaptor mount 875 is connected to a motor tower that moves the spigot 872 up and down, and positions the spigot 872 so it can deliver fluid to each culture plate one at a time. When the controller has moved the spigot 872 into a position for delivery of fluid to a specific plate, the controller moves the spigot forward so that the front 876 of the spigot contacts the slidable flap on a culture plate and moves the flap from the closed position to the open position to allow deli very of the fluid from the spigot 872 to the culture plate. At this point, fluid is pumped through the tube 871, raising the diaphragm 873 and fluid enters the plate through the fluid inlet.

[00173] Figure 9 shows an elevated perspective view of a tower 980 holding two assemblies 950 for use in an incubator. Each assembly is supported by two shelf rails 981 , which allow slidable entry of the assemblies into the tower 980. The assemblies 950 each hold a plurality of culture plates 910, the fluid outlets 919 of which are connected to a waste container 955 to manage fluid removed from the plates 910 by tilting.

[00174] The tower 980 sits in a base board 982, which has height adjustable feet 983.

[00175] The tower 980 comprises a motor tower 984 for controlling the height of the spigot 970, which engages with a slidable flap 920 on each plate. An additional motor 985 controls forward and backward motion of the spigot 970.

[00176] Tilting of the culture plates 910 is achieved by positioning the spigot 970 between culture plates 910 and then controlling the height of the spigot 970 with motor 984 so that the top of the spigot 970 lifts the plate above it a predetermined distance, and tilt the plates of each assembly 950.

[00177] Figure 10 shows an elevated perspective view of two towers 1080 and a media reservoir assembly 1085 for supplying fluid (eg a culture medium) to the culture plates 1010 according to one embodiment. Each tower 1080 comprises a separate controlled motor system to deliver fluid and tilt the plates in each assembly 1050, using motor towers 1084.

[00178] The medium reservoir assembly 1085 comprises one or more bags 1086 filled with fluid and having fluid delivery tubes 1087 connected to the fluid delivery means for the plates.

[00179] Figure 1 1 shows an elevated perspective view of assembly (a bioreactor) 1 190 according to one embodiment, the assembly 1 190 comprising an incubator 1 191 for providing suitable culture conditions, two towers 1 180 each comprising two assemblies 1 150 for providing fluid management of culture plates 11 10, motor towers 1184, and medium reservoir assembly 1 185.

[00180] In addition to the assembly 1 190, there is a media fridge 1 192 and a pump and motor drive housing 1 193, which additionally has inside it both the power supply for the pumps and motors, and the electronic circuitry to deliver power to the pumps and motors. Also not shown is a computer which hosts the custom control software that provides control instructions to the pump and motor drive housing.

[00181 ] Although the present disclosure has been described with reference to particular embodiments, it will be appreciated that the disclosure may be embodied in many other forms. It will also be appreciated that the disclosure described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the disclosure includes all such variations and modifications. The disclosure also includes all of the steps, features, compositions and compounds referred to, or indicated in this specification, individually or collectively, and any and all combinations of any two or more of the steps or features.

[00182] Also, it is to be noted that, as used herein, the singular forms "a", "an" and "the" include plural aspects unless the context already dictates otherwise.

[00183] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers.

[00184] Reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that this prior art forms part of the common general knowledge in any country.

[00185] The subject headings used herein are included only for the ease of reference of the reader and should not be used to limit the subject matter found throughout the disclosure or the claims. The subject headings should not be used in construing the scope of the claims or the claim limitations.

[00186] The description provided herein is in relation to several embodiments which may share common characteristics and features. It is to be understood that one or more features of one embodiment may be combinable with one or more features of the other embodiments. In addition, a single feature or combination of features of the embodiments may constitute additional embodiments.

[00187] All methods described herein can be performed in any suitable order unless indicated otherwise herein or clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illuminate the example embodiments and does not pose a limitation on the scope of the claimed invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential.

[00188] Future patent applications may be filed on the basis of the present application, for example by claiming priority from the present application, by claiming a divisional status and/or by claiming a continuation status. It is to be understood that the following claims are provided by way of example only, and are not intended to limit the scope of what may be claimed in any such future application. Nor should the claims be considered to limit the understanding of (or exclude other understandings of) the present disclosure. Features may be added to or omitted from the example claims at a later date.

[00189] Although the present disclosure has been described with reference to particular examples, it will be appreciated by those skilled in the art that the disclosure may be embodied in many other forms.

Claims

1. An assembly for culturing cells or tissue, the assembly comprising:

wherein the waste container is held within the culture vessel holder.

2. The assembly according to claim 1, wherein the assembly comprises a manifold for fluid connection of the fluid outlet of one or more of the culture vessels to the waste container.

3. The assembly according to claim 2, wherein the manifold is removably connected to the fluid outlet.

4. The assembly according to claims 2 or 3, wherein the manifold is removably connected to the waste container and/or a hose connected thereto.

5. The assembly according to any one of claims 1 to 4, wherein the culture vessel holder is configured to hold a culture vessel having a top and/or a bottom surface area of greater than 400 cm².

6. The assembly according to any one of claims 1 to 5, wherein the culture vessel holder holds 5 or more culture vessels.

7. The assembly according to any one of claims 1 to 6, wherein the waste container is located below the level of the culture vessels.

8. The assembly according to claim 7, wherein the waste container is located at the base of the culture vessel holder.

9. The assembly according to any one of claims 1 to 8, wherein the waste container is removable from the culture vessel holder.

10. The assembly according to any of claims 1 to 9, wherein the waste container is a replaceable component.

1 1. The assembly according to any one of claims 1 to 10, wherein the fluid inlet and the fluid outlet are located substantially opposed to one another.

12. The assembly according to any one of claims 1 to 1 1, wherein the holder is configured to pennit tilting of the culture vessels in the holder so as to remove the fluid from the culture vessels via the fluid outlet upon tilting of the culture vessels.

13. The assembly according to claim 12, wherein the culture vessel holder comprises a means for supporting the culture vessels and the means for supporting is pivotally connected to the holder to permit tilting of the culture vessels.

14. The assembly according to claims 12 or 13, wherein the tilting of the culture vessels is biased towards the horizontal position.

15. The assembly according to claim 14, wherein the assembly comprises a spring to bias the tilting of the culture vessels towards the horizontal position.

16. The assembly according to any one of claims 13 to 15, wherein the means for supporting culture vessels comprises one or more vertical panels with ledges.

17. The assembly according to claim 16, wherein the panels comprise laterally displaced front tabs to assist with guiding the culture vessels into the culture vessel holder and/or prevent the culture vessels from being withdrawn.

18. The assembly according to any one of claims 1 to 17, wherein the culture vessel holder holds one or more culture vessels.

19. The assembly according to any one of claims 1 to 18, wherein the culture vessels comprise a culture plate or a culture flask.

20. An assembly for culturing cells or tissue, the assembly comprising:

wherein the waste container is held within the culture vessel holder.

21. A method of controlling fluid exchange in a culture vessel, the method including using an assembly according to any one of claim 1 to 20 to control fluid exchange in the culture vessel.

22. An incubator comprising an assembly according to any one of claims 1 to 20.

23. An incubator for culturing cells or tissue, the incubator comprising an assembly comprising:

wherein the waste container is held within the culture vessel holder.

24. The incubator according to claim 23, wherein the culture vessel holder holds one or more culture vessels.

25. The incubator according to claims 23 or 24, wherein the incubator comprises a plurality of assemblies.

26. The incubator according to any one of claims 23 to 25, wherein the assembly comprises a manifold for fluid connection of the fluid outlet of one or more of the culture vessels to the waste container.

27. The incubator according to claim 26, wherein the manifold is removably connected to the fluid outlet.

28. The incubator according to claims 26 or 27, wherein the manifold is removably connected to the waste container and/or a hose connected thereto.

29. The incubator according to any one of claims 23 to 28, wherein the culture vessel holder is configured to hold a culture vessel having a top and/or a bottom surface area of greater than 400 cm².

30. The incubator according to any one of claims 23 to 29, wherein the culture vessel holder holds 5 or more culture vessels.

31. The incubator according to any one of claims 23 to 30, wherein the waste container is removable from the culture vessel holder.

32. The incubator according to any one of claims 23 to 31 , wherein the fluid inlet and the fluid outlet are located substantially opposed to one another.

33. The incubator according to any one of claims 23 to 32, wherein the holder is configured to permit tilting of the culture vessels in the holder so as to remove fluid from the culture vessels via the fluid outlet upon tilting of the culture vessels.

34. The incubator according to claim 33, wherein the culture vessel holder comprises a means for supporting the culture vessels and the means for supporting is pivotally connected to the holder to permit tilting of the culture vessels.

35. The incubator according to claims 33 or 34, wherein the tilting of the culture vessels is biased towards the horizontal position.

36. The incubator according to claim 35, wherein the assembly comprises a resilient component and/or a spring to bias the tilting of the culture vessels towards the horizontal position.

37. The incubator according to any one of claims 23 to 36, wherein the incubator comprises a mechanical means to tilt the culture vessel.

38. The incubator according to any one of claims 23 to 37, wherein the incubator comprises a fluid input means to deliver fluid to the culture vessels through the fluid inlet.

39. The incubator according to any one of claims 23 to 38, wherein the fluid input means comprises a spigot.

40. The incubator according to claims 38 or 39, wherein the fluid input means is used to deliver fluid to a plurality of culture vessels.

41. The incubator according to any one of claims 38 to 40, wherein the fluid input means delivers fluid to individual culture vessels.

42. The incubator according to any one of claims 38 to 41 , wherein the fluid input means is mechanically controlled.

43. The incubator according to any one of claims 23 to 42, wherein the fluid inlet comprises a means for sealing the inlet.

44. The incubator according to any one of claims 38 to 43, wherein the fluid input means engages with the means for sealing the inlet to deliver fluid to the culture vessel.

45. The incubator according to claims 43 or 44, wherein the means for sealing the fluid inlet comprises a moveable flap.

46. The incubator according to claim 45, wherein the fluid input means moves the flap to an open position to deliver fluid and the flap returns to a sealed position upon withdrawal of the fluid input means.

47. The incubator according to any one of claims 23 to 46, wherein the incubator comprises one or more fluid reservoirs.

48. The incubator according to claim 47, wherein the one or more fluid reservoirs comprises a culture medium reservoir.

49. The incubator according to any one of claims 23 to 48, wherein the incubator comprises means to mechanically control delivery and/or removal of the fluid to the culture vessels.

50. The incubator according to any one of claims 23 to 49, wherein the incubator comprises computer controlled means to automatically deliver and/or remove fluid.

51. The incubator according to any one of claims 23 to 50, wherein the incubator provides computer controlled management of fluid exchange for the culture vessels.

52. The incubator according to any one of claims 23 to 51 , wherein the culture vessels comprise culture plates or culture flasks.

53. A culture vessel comprising a fluid inlet permitting fluid to enter the culture vessel and a fluid outlet pennitting fluid to exit the culture vessel.

54. The culture vessel according to claim 53, wherein the culture vessel comprises a culture plate with a base and a removable lid.

55. The culture vessel according to claim 54, wherein the lid comprises the fluid inlet.

56. The culture vessel according to claims 54 or 55, wherein the base comprises the fluid outlet.

57. The culture vessel according to any one of claims 53 to 56, wherein the culture vessel comprises a means to reduce fluid velocity of fluid entering the culture vessel through the fluid inlet.

58. The culture vessel according to claim 57, wherein the means to reduce fluid velocity comprises one or more surfaces above the level of cell or tissue culturing and/or one or more baffles.

59. The culture vessel according to claims 57 or 58, wherein the means to reduce fluid velocity comprise one or more barriers.

60. The culture vessel according to any one of claims 53 to 59, wherein the culture vessel comprises one or more internal barriers to hold a desired level of fluid in the culture vessel.

61. The culture vessel according to claim 60, wherein the one or more internal barriers prevent fluid from flowing out the fluid outlet.

62. The culture vessel according to claim 61 , wherein one of the internal barriers comprises one or more recesses below the level of the top of the other internal barriers.

63. The culture vessel according to any one of claims 53 to 62, wherein the fluid inlet comprises a means for sealing the fluid inlet.

64. The culture vessel according to claim 63, wherein the means for sealing the fluid inlet comprises a moveable flap.

65. The culture vessel according to claim 64, wherein movement of the moveable flap from a sealed position to an open position permits delivery of fluid through the fluid inlet.

66. The culture vessel according to claim 65, wherein the moveable flap comprises a resilient means to return the moveable flap from the open position to the sealed position.

67. The culture vessel according to any one of claims 64 to 66, wherein the moveable flap is a slidable flap.

68. The culture vessel according to any one of claims 53 to 67, wherein the culture vessel comprises a removable platform for growing cells or tissue on the platform or for growing cells or tissue on a scaffold supported by the removable platform.

69. The culture vessel according to claim 68, wherein the culture vessel comprises a culture plate with a base and a removable lid and the removable lid comprises one or more projections to hold the removable platform under fluid in use.

70. A culture vessel comprising a removable platform for growing cells or tissue on the platform or for growing cells or tissue on a scaffold supported by the removable platform.

71. The culture vessel according to claim 70, wherein the culture vessel comprises a culture plate with a base and a removable lid.

72. The culture vessel according to claim 71, wherein the removable lid comprises one or more projections to hold the removable platform under fluid in use.

73. The culture vessel according to any one of claims 70 to 72, wherein the culture vessel comprises a fluid inlet permitting fluid to enter the culture vessel and a fluid outlet permitting fluid to exit the culture vessel.

74. The culture vessel according to claim 73, wherein the culture vessel comprises a culture plate with a base and a removable lid and the lid comprises the fluid inlet.

75. The culture vessel according to claims 73 or 74, wherein the culture vessel comprises a culture plate with a base and a removable lid and the base comprises the fluid outlet.

76. The culture vessel according to any one of claims 73 to 75, wherein the culture vessel comprises a means to reduce fluid velocity entering the culture vessel through the fluid inlet.

77. The culture vessel according to claim 76, wherein the means to reduce fluid velocity comprises one or more surfaces above the level of cell or tissue culturing and/or one or more baffles.

78. The culture vessel according to claims 76 or 77, wherein the means to reduce fluid velocity comprise one or more barriers.

79. The culture vessel according to any one of claims 70 to 78, wherein the culture vessel comprises one or more internal barriers to hold a desired level of fluid in the culture vessel.

80. The culture vessel according to claim 79, wherein the one or more barriers prevent fluid from flowing out a fluid outlet,

81. The culture vessel according to claim 80, wherein a barrier comprises one or more recesses below the level of the top of the other internal barriers.

82. The culture vessel according to any one of claims 74 to 81 , wherein the fluid inlet comprises a means for sealing the fluid inlet.

83. The culture vessel according to claim 82, wherein the culture vessel comprises a moveable flap to seal the fluid inlet.

84. The culture vessel according to claim 83, wherein movement of the moveable flap from a closed position to an open position permits delivery of fluid through the fluid inlet.

85. The culture vessel according to claim 84, wherein the moveable flap comprises a resilient means to return the moveable flap from the open position to a closed position.

86. The culture vessel according to any one of claims 83 to 85, wherein the moveable flap is a slidable flap.

87. A method of culturing cells or tissue, the method comprising incubating the cells or tissue in an incubator according to any one of claims 22 to 53.

88. A method of culturing cells or tissue, the method comprising culturing cells or tissue in a culture vessel according to any one of claims 53 to 86.

89. A bioreactor for culturing cells or tissue, the bioreactor comprising an assembly according to any one of claims 1 to 20, an incubator and a fluid delivery means for delivering fluid to the assembly.