Highly durable permeable fluoropolymer cell culture bag

Abstract

To provide a highly durable permeable fluoropolymer cell culture bag.SOLUTION: A cell culture bag includes a body formed of a composite film including a first fluoropolymer and a second fluoropolymer. The body defines a cell culture compartment configured to hold a cell culture. The first fluoropolymer has a first thickness and the second fluoropolymer at least partially impregnates the first thickness of the first fluoropolymer. The composite film has a second thickness from 0.01 mm to 0.059 mm and tensile strength in one direction of 10,000 psi to 92,000 psi. The composite film also has an O2 flux of 2,000 cm3 / m2 / atm / day to 20,000 cm3 / m2 / atm / day and total transmittance from 70% to 100%. A cell culture assembly includes a plurality of the cell culture bags connected in series. A cell culture container includes a cell culture compartment and a composite film connected to the cell culture compartment.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] FIELD OF THE DISCLOSURE The present disclosure relates generally to gas permeable materials or components, and more specifically to fluoropolymer cell culture bags having non-polar gas permeability. [Background technology]

[0002] Cell and gene therapy are increasingly viable methods for treating many conditions, including various cancers, neurological disorders, infectious diseases such as tuberculosis and cystic fibrosis, ulcerative colitis, peripheral arterial disease, aneurysms, heart disease, Alzheimer's and Parkinson's diseases, autism, ophthalmologic conditions, diabetes, and other medical conditions. Generally, with respect to cell and gene therapy, various cell types can be grown in vitro. In vitro cell culture is a complex process in which cells are grown under controlled conditions outside their natural environment but that are as close as possible to their natural in vivo conditions.

[0003] One method of culturing cells in vitro is by using cell culture bags, such as fluoropolymer cell culture bags, which reduce the risk of cell culture contamination because the cell culture bags are disposable and provide a closed system. However, some current fluoropolymer cell culture bags lack the durability to function reliably as cell culture bags. Specifically, current fluoropolymer cell culture bags have a tensile strength that can cause them to break during handling. The durability of these cell culture bags can be increased by increasing the thickness of the fluoropolymer film used to form the cell culture bag. However, increasing the film thickness can result in reduced gas permeation of gases, such as both oxygen (O2) and carbon dioxide (CO2), which are necessary for cultured cells to survive. Therefore, there is a need for cell culture bags that can withstand the stresses of handling while providing an environment suitable for cell metabolism and growth.

[0004] Another existing method for culturing cells in vitro is to use cell culture vessels, such as flasks, that contain fluoropolymer films that are permeable to gases such as O and CO. Current cell culture vessels also contain fluoropolymer composite films with tensile strength that can break during handling. Furthermore, current cell culture vessels can contain "dirty" films, e.g., composite films that contain undesirable amounts of extractables and / or leachables. Furthermore, conventional fluoropolymer composite films do not provide the transparency necessary to view the interior of the cell culture vessel. Therefore, there is a need for cell culture vessels that can withstand the strain of handling while allowing visibility within the vessel and providing an environment suitable for cell metabolism and growth. Summary of the Invention

[0005] This Summary is a high-level overview of various aspects of the invention and introduces some concepts that are described in more detail in the Detailed Description section below. This Summary is not intended to identify key or essential features of the claimed subject matter, nor is it intended to be used alone to determine the scope of the claimed subject matter. The subject matter should be understood by reference to the entire specification, some or all of the drawings, and appropriate portions of each claim.

[0006] Embodiments of the present disclosure relate to, but are not limited to, a cell culture bag comprising a body formed from a composite film including a first fluoropolymer and a second fluoropolymer, the body defining a cell culture compartment configured to hold a cell culture. The first fluoropolymer has a first thickness, and the second fluoropolymer at least partially penetrates the first thickness of the first fluoropolymer. The composite film has a second total thickness of 0.01 mm to 0.059 mm, where the second total thickness is the combined thickness of the first fluoropolymer and the second fluoropolymer. The composite film has a unidirectional tensile strength of 10,000 psi to 92,000 psi. The composite film has an O2 flux of 2,000 cm 3 / m 2 / atm / day~20,000cm 3 / m 2 / atm / day, with a total transmittance of 70% to 100%.

[0007] In some embodiments, the composite film is silicone-free.

[0008] In some embodiments, the second fluoropolymer penetrates between 0.00001 mm and 0.02 mm of the first thickness of the first fluoropolymer.

[0009] In some embodiments, the composite film has a total thickness of 0.02 mm to 0.059 mm.

[0010] In some embodiments, the composite film has a total organic carbon (TOC) in water of 0.00001 mg / cm 2 ~1mg / cm 2 is.

[0011] In some embodiments, the composite film has an O flux of 8,000 cm 3 / m 2 / atm / day~15,000cm 3 / m 2 / atm / day.

[0012] In some embodiments, the composite film has a tensile strength in at least one direction of from 20,000 psi to 92,000 psi.

[0013] In some embodiments, the peel strength of the composite film is 0.25 N / mm to 10 N / mm.

[0014] In some embodiments, the first fluoropolymer is expanded polytetrafluoroethylene and the second fluoropolymer is fluorinated ethylene propylene.

[0015] Embodiments of the present disclosure relate to, but are not limited to, cell culture bags including a tube extending from a first end to a second end and defining a cell culture compartment. The tube is formed from a first composite film including a first fluoropolymer and a second fluoropolymer. The first fluoropolymer has a first thickness, and the second fluoropolymer at least partially penetrates the first thickness of the first fluoropolymer. The first composite film has a second thickness of 0.01 mm to 0.059 mm. The first composite film has an O2 flux of 2,000 cm3 or greater. 3 / m 2 / atm / day~20,000cm 3 / m 2 / atm / day. The first composite film has a tensile strength of 10,000 psi to 92,000 psi. The first composite film has a total permeability of at least 70%. The cell culture bag also includes a first seam at a first end, a second composite film configured to fold over the first seam to form a first lap seam over the first seam, and a second lap seam extending longitudinally from the first end to a second end, the second lap seam being formed by overlapping edges of the first composite film.

[0016] In some embodiments, the cell culture bag further includes a second seam at a second end and a third composite film configured to fold over the second seam to form a third lap seam.

[0017] In some embodiments, the second fluoropolymer penetrates between 0.00001 mm and 0.005 mm of the first thickness of the first fluoropolymer.

[0018] In some embodiments, the first composite film has a thickness of 0.02 mm to 0.059 mm.

[0019] In some embodiments, the first composite film has a total organic carbon (TOC) of 1 mg / cm2 is less than.

[0020] In some embodiments, the first composite film has an O flux of 8,000 cm 3 / m 2 / atm / day~15,000cm 3 / m 2 / atm / day.

[0021] In some embodiments, the first composite film has a tensile strength in at least one direction of 20,000 psi to 92,000 psi.

[0022] In some embodiments, the first fluoropolymer is expanded polytetrafluoroethylene and the second fluoropolymer is fluorinated ethylene propylene.

[0023] In some embodiments, the first fluoropolymer is densified expanded polytetrafluoroethylene and the second fluoropolymer is fluorinated ethylene-propylene.

[0024] In some embodiments, the cell culture bag further comprises at least one access port in fluid communication with the cell culture compartment.

[0025] In some embodiments, the cell culture comprises at least one of connective tissue cells, skeletal cells, cardiac cells, epithelial cells, neural cells, endocrine cells, immune cells, lymphocytes, melanocytes, tumor cells, or a combination thereof.

[0026] In some embodiments, the first composite film and the second composite film are the same.

[0027] In some embodiments, the third composite film is the same as at least one of the first composite film or the second composite film.

[0028] In some embodiments, the peel strength of the first composite film is 0.25 N / mm to 10 N / mm.

[0029] Embodiments of the present disclosure also relate to, but are not limited to, cell culture assemblies including a plurality of cell culture bags connected in series. Each of the cell culture bags includes a body formed from a composite film including a first fluoropolymer and a second fluoropolymer, the body defining a cell culture compartment configured to hold a cell culture. The first fluoropolymer has a first thickness, and the second fluoropolymer at least partially penetrates the first thickness of the first fluoropolymer. The composite film has a second thickness of 0.01 mm to 0.059 mm. The composite film has a unidirectional tensile strength of 10,000 psi to 92,000 psi. The composite film has an O2 flux of 2,000 cm 3 / m 2 / atm / day~20,000cm 3 / m 2 / atm / day, and the total transmittance of the composite film is 70% to 100%.

[0030] In some embodiments, a first cell culture bag of the plurality of cell culture bags has a first O2 flux and a second cell culture bag of the plurality of cell culture bags has a second O2 flux.

[0031] In some embodiments, the first O2 flux and the second O2 flux are different.

[0032] In some embodiments, a first cell culture bag of the plurality of cell culture bags has a first volume and a second cell culture bag of the plurality of cell culture bags has a second volume.

[0033] In some embodiments, the first volume is different from the second volume.

[0034] In some embodiments, a first cell culture bag of the plurality of cell culture bags is configured for at least one of cell transfection or cell activation.

[0035] In some embodiments, a second cell culture bag of the plurality of cell culture bags is configured for cell expansion.

[0036] Embodiments of the present disclosure also relate to a cell culture vessel including a cell culture compartment configured to receive a cell culture. The cell culture vessel also includes a composite film connected to the cell culture compartment, the composite film including a first fluoropolymer and a second fluoropolymer. The first fluoropolymer has a first thickness, and the second fluoropolymer at least partially penetrates the first thickness of the first fluoropolymer. The composite film has a second thickness of 0.01 mm to 0.059 mm. The composite film has a unidirectional tensile strength of 10,000 psi to 92,000 psi. The composite film has an O2 flux of 2,000 cm 3 / m 2 / atm / day~20,000cm 3 / m 2 / atm / day, and the total transmittance of the composite film is 70% to 100%. [Brief explanation of the drawings]

[0037] The accompanying drawings are included to provide a further understanding of the disclosure, and are incorporated into and constitute a part of this specification, illustrate embodiments and, together with the description, serve to explain the principles of the disclosure.

[0038] [Figure 1] FIG. 1 is a top view of a cell culture bag according to some embodiments herein.

[0039] [Figure 2] FIG. 2 is a perspective view of a composite film tube of a cell culture bag according to some embodiments herein.

[0040] [Figure 3] FIG. 3 is a perspective view of four cell culture bags connected in series, according to some embodiments herein.

[0041] [Figure 4] FIG. 4 is a front view of a cell culture vessel according to some embodiments herein. DETAILED DESCRIPTION OF THE INVENTION

[0042] Those skilled in the art will readily appreciate that the various aspects of the present disclosure may be implemented by any number of methods and apparatus configured to perform the intended functions. It should also be noted that the accompanying drawings referred to herein are not necessarily drawn to scale and may be exaggerated to illustrate various aspects of the present disclosure, and in that regard the drawings should not be construed as limiting.

[0043] Described herein are cell culture bags and vessels comprising composite films for in vitro culturing of cells. Also described herein are methods for forming the composite films used to form the cell culture bags and vessels. In some embodiments, the composite films have an O flux that promotes cell metabolism and growth within the cell culture vessel. Furthermore, in some embodiments, the composite films also have sufficient tensile strength to reduce or even prevent tearing or leakage of the cell culture bag or vessel during handling. In some embodiments, the composite films are densified. The cell culture bags and vessels described herein can be useful for treating cells from patients with pathological diseases or conditions in which cell loss or insufficient growth of cultured cells in cell culture can affect the success of the treatment.

[0044] 1 and 2 each illustrate a cell culture bag 100 according to some embodiments of the present disclosure. The cell culture bag 100 is formed from a composite film tube 104 defining a cell culture compartment 114 therein, a first end 106, and a second end 108. The first end 106 is a handle end, while the second end 108 is a port end. In one embodiment, the second end 108 has at least one sealable port 112 and port adapter 110 in fluid communication with the cell culture compartment 114 for inserting, removing, sampling, and delivering a cell culture 116 therein.

[0045] The composite film tube 104 is formed from at least one composite film 102. A "composite film" is defined herein as a film formed from at least two different materials or films. For example, in some embodiments, the composite film 102 can be formed by layering at least two materials or films and applying heat and / or pressure such that the layers of the resulting composite film 102 are inextricably bonded to one another.

[0046] In one embodiment, the composite film 102 is densified generally according to the teachings provided in US Pat. No. 7,521,010 to Kennedy et al., which is incorporated herein by reference.

[0047] In some embodiments, composite film 102 comprises a non-polar gas-permeable fluoropolymer composite film. In some embodiments, cell culture bag 100 comprises a gas-permeable fluoropolymer composite film because the fluoropolymer composite film is biologically, chemically, and immunologically compatible with cell cultures.

[0048] In some embodiments, the composite film 102 of the cell culture bag 100 can include, but is not limited to, the following materials: fluorinated ethylene-propylene (FEP), tetrafluoroethylene (TFE), modified polytetrafluoroethylene, perfluoroalkoxy (PFA), polyvinyl fluoride (PVF), polychlorotrifluoroethylene (PCTFE), polyethylene tetrafluoroethylene (ETFE), chlorotrifluoroethylene vinylidene fluoride (FPM / FKM), polyethylene chlorotrifluoroethylene (ECTFE), perfluoroelastomers (FFPM / FFLM), perfluoropolyethers (PFPE), tetrafluoroethylene and perfluoromethyl vinyl ether copolymer (MFA), chlorotrifluoroethylene vinylidene fluoride copolymer (FTFE / VDF), and any combination thereof.

[0049] For ease of discussion, reference will be made herein to expanded polytetrafluoroethylene (ePTFE), but it should be understood that expanded modified polytetrafluoroethylene (PTFE), expanded blends of PTFE, expanded copolymers of PTFE, and PTFE homopolymers are all considered within the scope of the present invention. Patents have been issued relating to expandable blends of PTFE, expandable modified PTFE, and expanded copolymers of PTFE, such as U.S. Patent No. 5,708,044 to Branca, U.S. Patent No. 6,541,589 to Baillie, U.S. Patent No. 7,531,611 to Sabol et al., U.S. Patent No. 8,647,144 to Ford, and U.S. Patent No. 9,139,669 to Xu et al.

[0050] In some embodiments, the material for the composite film 102 comprises expanded polytetrafluoroethylene (ePTFE).

[0051] In some embodiments, the material for the composite film 102 comprises densified expanded polytetrafluoroethylene (ePTFE).

[0052] In some embodiments, the films and / or materials for the composite film 102 are silicone-free.

[0053] In some embodiments, the composite film 102 includes a first material and a second material. In some embodiments, each of the first material and the second material is a fluoropolymer. In some embodiments, the first material is ePTFE and the second material is FEP. In some embodiments, the composite film 102 is made generally according to the teachings provided in U.S. Patent No. 7,521,010 to Kennedy et al., which is incorporated herein by reference.

[0054] In some embodiments, the first material is at least partially penetrated by the second material to form a highly durable mechanical bond. Specifically, in some embodiments, the second material extends at least partially through the thickness of the first material. In some embodiments, the second material penetrates into the first material by 0.00001 mm to 0.02 mm. In other embodiments, the second material penetrates into the first material by 0.00005 mm to 0.02 mm. In other embodiments, the second material penetrates into the first material by 0.0001 mm to 0.02 mm. In other embodiments, the second material penetrates into the first material by 0.0005 mm to 0.02 mm. In other embodiments, the second material penetrates into the first material by 0.001 mm to 0.02 mm. In other embodiments, the second material penetrates into the first material by 0.005 mm to 0.02 mm. In another embodiment, the second material penetrates into the first material by 0.01 mm to 0.02 mm.

[0055] In some embodiments, the second material penetrates into the first material by 0.00001 mm to 0.01 mm. In other embodiments, the second material penetrates into the first material by 0.00001 mm to 0.005 mm. In other embodiments, the second material penetrates into the first material by 0.00001 mm to 0.001 mm. In other embodiments, the second material penetrates into the first material by 0.00001 mm to 0.0005 mm. In other embodiments, the second material penetrates into the first material by 0.00001 mm to 0.0001 mm. In other embodiments, the second material penetrates into the first material by 0.00001 mm to 0.0001 mm. In other embodiments, the second material penetrates into the first material by 0.00001 mm to 0.00005 mm.

[0056] In some embodiments, the second material penetrates into the first material by 0.0001 mm to 0.005 mm. In other embodiments, the second material penetrates into the first material by 0.0005 mm to 0.001 mm. In other embodiments, the second material penetrates into the first material by 0.00005 mm to 0.005 mm. In other embodiments, the second material penetrates into the first material by 0.001 mm to 0.005 mm. In other embodiments, the second material penetrates into the first material by 0.005 mm to 0.01 mm.

[0057] In some embodiments, the composite film 102 comprises a first material infiltrated with a second material on both a first side and a second side of the first material.

[0058] In some embodiments, the composite film 102 comprises a first material infiltrated on one side (e.g., the first side) of the first material with a second material and an opposite side (e.g., the second side) of the first material with a third material.

[0059] A characteristic of the composite film 102 for growing cell cultures is the total organic carbon (TOC) in the water. As defined herein, TOC is the amount of carbon bound in organic compounds and is often used as a non-specific indicator of water quality or the cleanliness of pharmaceutical equipment, among other things. TOC is used as a process control attribute in the biotechnology industry to monitor the performance of unit operations employing purification and distribution systems.

[0060] In some embodiments, the composite film 102 has a density of 0.0005 mg / cm 2 In other embodiments, the composite film has a TOC of less than 0.00001 mg / cm 2 ~1mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.00005 mg / cm 2 ~1mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.0001 mg / cm 2 ~1mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.005 mg / cm 2 ~1mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.01 mg / cm 2 ~1mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.1 mg / cm 2 ~1mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.5 mg / cm 2 ~1mg / cm 2 It has a TOC of

[0061] In some embodiments, the composite film 102 has a density of 0.00001 mg / cm 2 ~0.5mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.00001 mg / cm 2 ~0.1mg / cm 2In another embodiment, the composite film 102 has a TOC of 0.00001 mg / cm 2 ~0.05mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.00001 mg / cm 2 ~0.01mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.00001 mg / cm 2 ~0.005mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.00001 mg / cm 2 ~0.001mg / cm 2 In another embodiment, the composite film 102 has a TOC of 0.00001 mg / cm 2 ~0.0005mg / cm 2 It has a TOC of

[0062] In some embodiments, the composite film 102 has a density of 0.00005 mg / cm 2 ~0.005mg / cm 2 In some embodiments, the composite film 102 has a TOC of 0.0008 mg / cm 2 ~0.02mg / cm 2 In some embodiments, the composite film 102 has a TOC of 0.006 mg / cm 2 ~0.4mg / cm 2 In some embodiments, the composite film 102 has a TOC of 0.04 mg / cm 2 ~0.8mg / cm 2 In some embodiments, the composite film 102 has a TOC of 0.5 mg / cm 2 ~0.75mg / cm 2 In some embodiments, the composite film 102 has a TOC of 0.00005 to 0.0001 mg / cm 2 It has a TOC of

[0063] In some embodiments, the composite film 102 has a tensile strength of 10,000 psi (69 MPA) or greater in at least one direction. In another embodiment, the tensile strength of the composite film 102 in at least one direction is 12,000 psi or greater. In another embodiment, the tensile strength of the composite film 102 in at least one direction is 15,000 psi or greater. In another embodiment, the tensile strength of the composite film 102 in at least one direction is 17,500 psi or greater. In another embodiment, the tensile strength of the composite film 102 in at least one direction is 20,000 psi or greater. In another embodiment, the tensile strength of the composite film 102 in at least one direction is 35,000 psi or greater. In another embodiment, the tensile strength of the composite film 102 in at least one direction is 50,000 psi or greater. In another embodiment, the tensile strength of the composite film 102 in at least one direction is 60,000 psi or greater. In another embodiment, the tensile strength of the composite film 102 is 75,000 psi or greater in at least one direction. In another embodiment, the tensile strength of the composite film 102 is 80,000 psi or greater in at least one direction. In another embodiment, the tensile strength of the composite film 102 is 92,000 psi or less in at least one direction.

[0064] In some embodiments, the composite film 102 has a tensile strength in at least one direction of 10,000 psi to 92,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 10,000 psi to 80,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 10,000 psi to 60,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 10,000 psi to 50,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 10,000 psi to 25,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 10,000 psi to 20,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 10,000 psi to 15,000 psi. In another embodiment, the composite film 102 has a tensile strength in at least one direction of between 10,000 psi and 12,000 psi.

[0065] In some embodiments, the composite film 102 has a tensile strength in at least one direction of 12,000 psi to 92,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 15,000 psi to 92,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 20,000 psi to 92,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 25,000 psi to 92,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 40,000 psi to 92,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 50,000 psi to 92,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 75,000 psi to 92,000 psi.

[0066] In some embodiments, the composite film 102 has a tensile strength in at least one direction of 12,000 psi to 75,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 25,000 psi to 60,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 15,000 psi to 45,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 50,000 psi to 80,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 45,000 psi to 65,000 psi. In other embodiments, the composite film 102 has a tensile strength in at least one direction of 60,000 psi to 92,000 psi.

[0067] As defined herein, thickness is the dimension extending in the z-direction from a first surface of the composite film 102 to a second surface of the composite film 102. In some embodiments, the composite film 102 has a thickness of 0.01 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.01 mm to 0.09 mm. In other embodiments, the composite film 102 has a thickness of 0.01 mm to 0.085 mm. In other embodiments, the composite film 102 has a thickness of 0.01 mm to 0.075 mm. In other embodiments, the composite film 102 has a thickness of 0.01 mm to 0.065 mm. In other embodiments, the composite film 102 has a thickness of 0.01 mm to 0.059 mm. In other embodiments, the composite film 102 has a thickness of 0.01 mm to 0.05 mm. In other embodiments, the composite film 102 has a thickness of 0.01 mm to 0.04 mm. In another embodiment, the composite film 102 has a thickness of 0.01 mm to 0.03 mm. In another embodiment, the composite film 102 has a thickness of 0.01 mm to 0.025 mm. In another embodiment, the composite film 102 has a thickness of 0.01 mm to 0.015 mm.

[0068] In some embodiments, the composite film 102 has a thickness of 0.015 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.025 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.03 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.045 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.05 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.059 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.065 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.075 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.09 mm to 0.1 mm. In other embodiments, the composite film 102 has a thickness of 0.095 mm to 0.1 mm.

[0069] In some embodiments, the composite film 102 has a thickness of 0.02 mm to 0.045 mm. In other embodiments, the composite film 102 has a thickness of 0.025 mm to 0.035 mm. In other embodiments, the composite film 102 has a thickness of 0.03 mm to 0.04 mm. In other embodiments, the composite film 102 has a thickness of 0.02 mm to 0.025 mm. In other embodiments, the composite film 102 has a thickness of 0.075 mm to 0.095 mm. In other embodiments, the composite film 102 has a thickness of 0.065 mm to 0.08 mm. In other embodiments, the composite film 102 has a thickness of 0.059 mm to 0.098 mm.

[0070] In some embodiments, the composite film 102 has a transparency that allows the cell culture 116 within the cell culture compartment 114 to be viewed through the composite film 102. As used herein, transparency is quantified as the percent total transmittance of the composite film. Specifically, a higher percent total transmittance indicates higher transparency, while a lower percent total transmittance indicates lower transparency. In some embodiments, the composite film 102 has a percent total transmittance of 70% to 100%. In other embodiments, the composite film 102 has a percent total transmittance of 75% to 100%. In other embodiments, the composite film 102 has a percent total transmittance of 80% to 100%. In other embodiments, the composite film 102 has a percent total transmittance of 85% to 100%. In other embodiments, the composite film 102 has a percent total transmittance of 90% to 100%. In other embodiments, the composite film 102 has a percent total transmittance of 95% to 100%.

[0071] In some embodiments, the composite film 102 has a total transmittance of 70% to 95%. In other embodiments, the composite film 102 has a total transmittance of 70% to 90%. In other embodiments, the composite film 102 has a total transmittance of 70% to 85%. In other embodiments, the composite film 102 has a total transmittance of 70% to 80%. In other embodiments, the composite film 102 has a total transmittance of 70% to 85%.

[0072] In some embodiments, the composite film 102 has a total transmittance of 75% to 85%. In other embodiments, the composite film 102 has a total transmittance of 90% to 95%. In other embodiments, the composite film 102 has a total transmittance of 80% to 95%. In other embodiments, the composite film has a total transmittance of 75% to 95%. In other embodiments, the composite film has a total transmittance of 80% to 90%. In other embodiments, the composite film has a total transmittance of 75% to 90%.

[0073] In some embodiments, reducing the thickness of the composite film 102 results in a cell culture bag 100 with non-polar gas permeability, allowing oxygen to flow to the cells to promote cell survival and proliferation. As defined herein, non-polar gas permeability is defined as an O flux of at least 2,000 cm 3 / m 2 / atm / day. For purposes of this provision, water vapor is not a non-polar gas.

[0074] O2 flux is defined herein as the amount of oxygen deliverable to the cell culture 116 through the composite film 102. In some embodiments, the composite film 102 has an O2 flux of 2,000 cm3 or greater. 3 / m 2 / atm / day~20,000cm 3 / m 2 / atm / day. In some embodiments, the composite film 102 has an O flux of 5,000 cm 3 / m 2 / atm / day~20,000cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 10,000 cm 3 / m 2 / atm / day~20,000cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 15,000 cm 3 / m 2 / atm / day~20,000cm 3 / m 2 / atm / day.

[0075] In some embodiments, the composite film 102 has an O flux of 2,000 cm 3 / m 2 / atm / day~15,000cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 2,000 cm 3 / m2 / atm / day~10,000cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 2,000 cm 3 / m 2 / atm / day~7,500cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 2,000 cm 3 / m 2 / atm / day~5,000cm 3 / m 2 In another embodiment, the composite film 102 has an O flux of 2,000 cm 3 / m 2 / atm / day~2,500cm 3 / m 2 / atm / day.

[0076] In some embodiments, the composite film 102 has an O flux of 3,000 cm 3 / m 2 / atm / day~8,000cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 6,000 cm 3 / m 2 / atm / day~16,000cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 4,500 cm 3 / m 2 / atm / day~5,000cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 15,000 cm 3 / m 2 / atm / day~17,500cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 12,000 cm 3 / m 2 / atm / day~18,000cm 3 / m 2 / atm / day. In another embodiment, the composite film 102 has an O flux of 17,500 cm 3 / m 2 / atm / day~19,000cm 3 / m 2 / atm / day.

[0077] As described herein, penetration of the second material into the first material creates a durable bond between the first and second materials of the composite film 102. In some embodiments, the durable bond between the first and second materials is sufficient to provide the composite film 102 with a peel strength of 0.25 N / mm to 10 N / mm. As defined herein, the peel strength of the composite film 102 is a measure of the average force required to separate (i.e., separate) the composite film 102 from the second film (or second material) to which it is bonded. As defined herein, peel strength can be the strength required to separate the ePTFE of the composite film 102 from the FEP of the second film, or the strength required to separate the FEP of the composite film 102 from the FEP of the second film. In some embodiments, the peel strength required to separate the ePTFE of the composite film 102 from the FEP of the second film is 0.25 N / mm to 10 N / mm. In another embodiment, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 0.5 N / mm to 10 N / mm. In another embodiment, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 1.0 N / mm to 10 N / mm. In another embodiment, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 2.5 N / mm to 10 N / mm. In another embodiment, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 5 N / mm to 10 N / mm. In another embodiment, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 7.5 N / mm to 10 N / mm.

[0078] In some embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 0.25 N / mm to 7.5 N / mm. In other embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 0.25 N / mm to 5 N / mm. In other embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 0.25 N / mm to 2.5 N / mm. In other embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 0.25 N / mm to 1.0 N / mm. In other embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 0.25 N / mm to 0.5 N / mm.

[0079] In some embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 0.5 N / mm to 7.5 N / mm. In other embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 1.0 N / mm to 5 N / mm. In other embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 2.5 N / mm to 7.5 N / mm. In other embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 1.0 N / mm to 2.5 N / mm. In other embodiments, the peel strength required to separate the ePTFE of composite film 102 from the FEP of the second film is 5 N / mm to 7.5 N / mm.

[0080] In some embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 0.25 N / mm to 10 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 0.5 N / mm to 10 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 1.0 N / mm to 10 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 2.5 N / mm to 10 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 5 N / mm to 10 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 7.5 N / mm to 10 N / mm.

[0081] In some embodiments, the peel strength of the FEP required to separate the FEP of composite film 102 from the FEP of the second film is 0.25 N / mm to 7.5 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 0.25 N / mm to 5 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 0.25 N / mm to 2.5 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 0.25 N / mm to 1.0 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 0.25 N / mm to 0.5 N / mm.

[0082] In some embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 0.5 N / mm to 7.5 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 1.0 N / mm to 5 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 2.5 N / mm to 7.5 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 1.0 N / mm to 2.5 N / mm. In other embodiments, the peel strength required to separate the FEP of composite film 102 from the FEP of the second film is 5 N / mm to 7.5 N / mm.

[0083] 1 and 2, the cell culture bag 100 is made by wrapping the composite film 102 around a heating element to form a tube. In some embodiments, a first end of the composite film 102 is overlapped with a second end of the composite film 102, and then the composite film 102 is bonded to itself using heat and pressure to create a composite film tube 104 having a longitudinal lap seam 118. The composite film tube 104 is then flattened in a direction perpendicular to the longitudinal axis L of the composite film tube 104.

[0084] Next, the second piece of composite film 130 is folded back onto itself. The first end 106 of the composite film tube 104 is placed between the two layers of the folded composite film 130. Heat and pressure are used to bond the folded composite film 130 to the composite film tube 104, creating a lap seam 132 that closes the first end 106 of the composite film tube 104. The folded composite film 130 is bonded to itself on the outside of the composite film tube 104 at a fin seam 142. Next, the third piece of composite film 140 is folded back onto itself. The second end 108 of the composite film tube 104 is placed between the two layers of the folded composite film 140. A non-adhesive element (not shown) is placed within the cell culture bag 100 extending into the folded composite film 140 to create an unbonded area. Heat and pressure are used to bond the folded composite film 140 to the composite film tube 104, creating a lap seam 144 that closes the second end of the composite film tube 104. The folded composite film 140 is bonded to itself on the outside of the composite film tube 104 at a fin seam 146. A slit is cut in the fin seam 146 of the cell culture bag 100. The non-adhesive elements are removed. A port adapter 110 is placed in the unbonded area. The port adapter 110 can be used in combination with various ports (not shown), depending on the desired end use. For example, in one embodiment, a non-collapsible port assembly can be used. Heat and pressure can be used to bond the port adapter 110 to the folded composite film 140, creating a fin seam (not shown), and forming the cell culture bag 100.

[0085] In yet another embodiment, the cell culture bag 100 is manufactured generally in accordance with the teachings provided in PCT Patent Application No. WO2019 / 209268 to Alford et al., which is incorporated herein by reference.

[0086] In other embodiments, other configurations of cell culture bag 100 can be used. For example, in some embodiments, cell culture bag 100 can be constructed generally in accordance with the teachings of U.S. Patent Application Publication No. 2016 / 0030283 A1 to Snyder et al.

[0087] In an alternative embodiment, two composite films 102 can be used, one composite film 102 on each side of the composite film tube 104 .

[0088] The volume inside the cell culture bag 100 can vary depending on the application. For example, in one embodiment, the cell culture bag 100 has a volume of 1 mL to 200 L. In another embodiment, the cell culture bag 100 has a volume of 100 mL to 200 L. In another embodiment, the cell culture bag 100 has a volume of 500 mL to 200 L. In another embodiment, the cell culture bag 100 has a volume of 1 L to 200 L. In another embodiment, the cell culture bag 100 has a volume of 5 L to 200 L. In another embodiment, the cell culture bag 100 has a volume of 10 L to 200 L. In another embodiment, the cell culture bag 100 has a volume of 50 L to 200 L. In another embodiment, the cell culture bag 100 has a volume of 100 L to 200 L.

[0089] In some embodiments, the cell culture bag 100 has a volume of 1 mL to 100 L. In other embodiments, the cell culture bag 100 has a volume of 1 mL to 50 L. In other embodiments, the cell culture bag 100 has a volume of 1 mL to 10 L. In other embodiments, the cell culture bag 100 has a volume of 1 mL to 5 L. In other embodiments, the cell culture bag 100 has a volume of 1 mL to 2 L. In other embodiments, the cell culture bag 100 has a volume of 1 mL to 1 L. In other embodiments, the cell culture bag 100 has a volume of 1 mL to 500 mL. In other embodiments, the cell culture bag 100 has a volume of 1 mL to 250 mL. In other embodiments, the cell culture bag 100 has a volume of 1 mL to 100 mL. In other embodiments, the cell culture bag 100 has a volume of 1 mL to 20 mL.

[0090] In some embodiments, the cell culture bag 100 has a volume of 1 mL to 150 L. In other embodiments, the cell culture bag 100 has a volume of 100 mL to 50 L. In other embodiments, the cell culture bag 100 has a volume of 500 mL to 1 L. In other embodiments, the cell culture bag 100 has a volume of 10 mL to 10 L. In other embodiments, the cell culture bag 100 has a volume of 5 L to 20 L. In some embodiments, the cell culture bag 100 has a volume of 1 L to 50 L.

[0091] In some embodiments, cell culture bag 100 is used for cell and gene therapy associated with many conditions including various cancers, neurological disorders, infectious diseases such as tuberculosis and cystic fibrosis, ulcerative colitis, peripheral arterial disease, aneurysms, heart disease, Alzheimer's and Parkinson's disease, autism, eye conditions, diabetes, and other medical conditions. In some embodiments, cell culture bag 100 is used for in vitro culture of cells during activation, transfection, expansion, and proliferation stages.

[0092] In some embodiments, at least two cell culture bags 100 are fluidly connected together in series, as shown in FIG. 3, to enable sterile transfer of cell cultures 116 during the cell culture process. In some embodiments, three cell culture bags are connected in series. In other embodiments, four cell culture bags are connected in series, as shown in FIG. 3. In other embodiments, five or more cell culture bags 100 can be connected in series. In some embodiments, the cell culture bags 100 in series have different characteristics based on the requirements of the cell cultures 116. For example, the size and O flux of each cell culture bag 100 can be adjusted based on the stage of the cell culture process. In some embodiments, two cell culture bags 100 with different sizes and O fluxes can be connected together. In other embodiments, three cell culture bags 100 with different sizes and O fluxes can be connected together. As one example, during stages where cell culture growth is required, the O flux of the composite film 102 can be increased compared to the O flux of the other cell culture bags 100. In other embodiments, the cell culture bag 100 can be connected to a freeze container bag, such as a Gore® STA-PURE™ Flexible Freeze Container, for sterile transport of the cell culture 116 for long-term storage anywhere from 2 months to 20 years or more.

[0093] In some embodiments, the interior or exterior surface of cell culture bag 100 is hydrophobic or includes a hydrophobic coating. In other embodiments, the interior or exterior surface of cell culture bag 100 is hydrophilic or includes a hydrophilic coating.

[0094] A variety of cell types can be grown in the cell culture bag 100, including, but not limited to, T cells, TCR cells, connective tissue cells, skeletal cells, cardiac cells, epithelial cells, neural cells, endocrine cells, immune cells, lymphocytes, and melanocytes. Similarly, a variety of growth media are available in which cell types can be grown, depending on the particular growth requirements and conditions of the cells. In some embodiments, suitable growth media include, for example, nutrient or lysogeny broth supplemented with hormones and / or growth factors.

[0095] In some embodiments, a coating or treatment is applied to the interior surface of the cell culture bag compartment 114 of the cell culture bag 100 to achieve a desired purpose. For example, the cell culture bag 100 can include a coating or treatment on the interior surface of the cell culture compartment 114 to promote cell growth and metabolism. In other embodiments, the coating or treatment can promote cell activation, transfection, expansion, and / or proliferation. However, any functional coating or treatment applied to the cell culture bag compartment 114 does not impede the O flux of the composite film 102. For example, in some embodiments, the coating or treatment is applied to the cell culture bag 100 in a pattern or on a portion of the interior surface of the cell culture compartment 114. In other embodiments, the composite film 102 is treated so that when the surface of the composite film 102 is treated, the resulting composite film 102 has a porosity of at least 2000 cm. 3 / m 2 / atm / day. In other embodiments, the composite film 102 is formed with an O flux above a threshold level, such that the cell culture bag 100 has an O flux of 100 / atm / day. In other embodiments, the composite film 102 is formed with an O flux, thickness, and tensile strength that allows the cell culture bag 100 to be functionalized after manufacture but before use. In some embodiments, the composite film 102 is formed with a surface area and / or surface energy that allows the cell culture bag 100 to be functionalized after manufacture but before use.

[0096] FIG. 4 illustrates a cell culture vessel 200 according to some embodiments of the present disclosure. The cell culture vessel 200 includes a cell culture compartment 214 configured to receive a cell culture 216. The cell culture vessel 200 also includes a composite film 202 disposed at or connected to the bottom of the cell culture vessel 200 and in contact with the cell culture 216 contained therein. The composite film 202 can be substantially the same as or identical to the composite film 102 described herein, including all of the properties of the composite film 102. In some embodiments, the composite film 202 is a fluoropolymer composite film that provides durability (e.g., tensile strength and puncture resistance) as well as the O flux and surface area necessary to support cell growth. Additionally, in some embodiments, the fluoropolymer composite film 202 reduces the amount of leachables and extractables.

[0097] The volume within the cell culture compartment 214 of the cell culture vessel 200 can vary depending on the desired application. For example, in one embodiment, the cell culture vessel 200 has a volume of 1 mL to 200 L. In another embodiment, the cell culture vessel 200 has a volume of 100 mL to 200 L. In another embodiment, the cell culture vessel 200 has a volume of 500 mL to 200 L. In another embodiment, the cell culture vessel 200 has a volume of 1 L to 200 L. In another embodiment, the cell culture vessel 200 has a volume of 10 L to 200 L. In another embodiment, the cell culture vessel 200 has a volume of 50 L to 200 L. In another embodiment, the cell culture vessel 200 has a volume of 100 L to 200 L.

[0098] In some embodiments, the cell culture vessel 200 has a volume of 1 mL to 100 L. In other embodiments, the cell culture vessel 200 has a volume of 1 mL to 50 L. In other embodiments, the cell culture vessel 200 has a volume of 1 mL to 10 L. In other embodiments, the cell culture vessel 200 has a volume of 1 mL to 1 L. In other embodiments, the cell culture vessel 200 has a volume of 1 mL to 750 mL. In other embodiments, the cell culture vessel 200 has a volume of 1 mL to 500 mL. In other embodiments, the cell culture vessel 200 has a volume of 1 mL to 100 mL. In other embodiments, the cell culture vessel 200 has a volume of 1 mL to 20 mL.

[0099] In some embodiments, the cell culture vessel 200 has a volume of 1 mL to 150 L. In other embodiments, the cell culture vessel 200 has a volume of 100 mL to 20 L. In other embodiments, the cell culture vessel 200 has a volume of 500 mL to 1 L. In other embodiments, the cell culture vessel 200 has a volume of 1 L to 2 L. In other embodiments, the cell culture vessel 200 has a volume of 2 L to 100 L. In other embodiments, the cell culture vessel 200 has a volume of 250 mL to 750 mL.

[0100] In some embodiments, changing the volume of cell culture vessel 200 results in a corresponding change in the surface area of ​​composite film 202 such that the surface area to volume ratio (SA / V) remains constant or nearly constant as the vessel volume changes to ensure adequate gas exchange for the cells contained therein.

[0101] In some embodiments, cell culture vessel 200 is used for cell and gene therapy associated with many conditions, including, but not limited to, various cancers, neurological disorders, infectious diseases such as tuberculosis and cystic fibrosis, ulcerative colitis, peripheral arterial disease, aneurysms, heart disease, Alzheimer's and Parkinson's disease, autism, ophthalmologic conditions, diabetes, and other medical conditions. In some embodiments, cell culture vessel 200 is used for in vitro culture of cells during activation, transfection, expansion, and proliferation stages. [Example]

[0102] Test Method O2 flux The O2 flux of the composite films described herein was measured according to ASTM D1434-82(2015).

[0103] tensile strength The tensile strength of the composite films described herein was measured according to ASTM D412-16 Die F.

[0104] Total organic carbon in water The TOC of the composite films described herein was measured using an apparatus that utilizes a high-temperature wet oxidation reaction of UV-assisted chemical oxidation in accordance with United States Pharmacopeia (USP) 643 (Ultra-Clean Technology Handbook: Volume 1: Ultra-Pure Water, Ohmi, Tadahiro; CRC Press, 1993, pp. 497-517). Purified water was added at 70°C for 24 hours, e.g., 3 cm per mL of water. 2 The composite film was contacted with a surface area ratio of 1:1. The water was removed from contact with the polymer and tested in a TOC analyzer. A suitable instrument strip is a Sievers M9 TOC analyzer from SUEZ WATER TECHNOLOGIES & SOLUTIONS.

[0105] Peel strength The peel strength of the composite films described herein was measured according to ASTM F88-15.

[0106] Total transmittance The average total transmittance through the composite films described herein was measured in accordance with ASTM D1003-13 for wavelengths between 380 and 740 nm.

[0107] Example 1 A 27.14 μm thick composite film containing 70 wt % ePTFE and 30 wt % FEP was obtained and was prepared generally according to U.S. Patent No. 7,521,010 to Kennedy.

[0108] The composite film had an FEP penetration thickness of 0.45 μm and an unpenetrated ePTFE thickness of 19.89 μm.

[0109] The composite film was tested for O2 flux using the test method set forth in ASTM D1434. The composite had an O2 flux of 11,500 cm 3 / m 2 It was determined that the daily average daily average was 1 / atm / day.

[0110] The composite film was also tested for tensile strength in at least one direction using the test method set forth in ASTM 412 Die F. The composite film was determined to have a minimum tensile strength of 10,520 psi in the machine and transverse directions.

[0111] The composite film exhibited a peel strength of 0.42 N / mm at the FEP / ePTFE interface and 1.12 N / mm at the FEP / FEP interface.

[0112] The total transmittance of the composite film was determined to be 91.0%.

[0113] Example 2 A 19.78 μm thick composite film containing 78 wt % ePTFE and 22 wt % FEP was obtained. The composite film was manufactured generally according to U.S. Patent No. 7,521,010 to Kennedy.

[0114] The composite film had an FEP penetration thickness of 0.35 μm and an unpenetrated ePTFE thickness of 14.86 μm.

[0115] The composite film was tested for O2 flux using the test method set forth in ASTM D1434. The composite film had an O2 flux of 7,786 cm 3 / m 2 It was determined that the daily average daily average was 1 / atm / day.

[0116] The composite film was also tested for tensile strength in at least one direction using the test method set forth in ASTM 412 Die F. The composite film was determined to have a minimum tensile strength of 24,100 psi in both the machine and transverse directions.

[0117] The composite had a peel strength of 0.43 N / mm at the FEP / ePTFE interface and 1.60 N / mm at the FEP / FEP interface.

[0118] The total transmittance of the composite film was determined to be 93.4%.

[0119] The results of Example 1 are shown in Table 1. [Table 1]

[0120] In comparison, conventional composite films were formed by adhering FEP to skived ePTFE. When tested, these composite films are expected to have FEP penetration, peel strength, tensile strength (machine and cross directions), and O flux outside the ranges provided in the examples and descriptions herein.

[0121] Additionally, conventional composite films formed by bonding FEP to ePTFE have been produced, and when tested, these conventional composite films are expected to have total transmittance values ​​outside the ranges provided in the examples and descriptions herein.

Claims

1. 1. A cell culture bag comprising a body formed from a composite film comprising a first fluoropolymer and a second fluoropolymer, the body defining a cell culture compartment configured to hold a cell culture; the first fluoropolymer has a first thickness; the second fluoropolymer at least partially penetrates a first thickness of the first fluoropolymer; the composite film has a second thickness of 0.01 mm to 0.059 mm; the composite film has a unidirectional tensile strength of 10,000 psi to 92,000 psi; The composite film is ２ Flux is 2,000 cm ３ / m ２ / atm / day ~ 20,000cm ３ / m ２ / atm / day, and The composite film has a total transmittance of 70% to 100%.

2. The cell culture bag of claim 1 , wherein the composite film is silicone-free.

3. 3. The cell culture bag according to claim 1, wherein the second fluoropolymer penetrates 0.00001 mm to 0.02 mm of the first thickness of the first fluoropolymer.

4. The cell culture bag according to any one of claims 1 to 3, wherein the composite film has a total thickness of 0.02 mm to 0.059 mm.

5. The composite film has a total organic carbon (TOC) content of 0.00001 mg / cm in water. ２ ~1 mg / cm ２ The cell culture bag according to any one of claims 1 to 4,

6. The composite film is ２ Flux is 8,000 cm ３ / m ２ / atm / day ~ 15,000cm ３ / m ２ The cell culture bag according to any one of claims 1 to 5, wherein the cell culture medium is a permeable medium.

7. The cell culture bag according to any one of claims 1 to 6, wherein the composite film has a tensile strength in at least one direction of 20,000 psi to 92,000 psi.

8. The cell culture bag according to any one of claims 1 to 7, wherein the peel strength of the composite film is 0.25 N / mm to 10 N / mm.

9. The cell culture bag according to any one of claims 1 to 8, wherein the first fluoropolymer is expanded polytetrafluoroethylene and the second fluoropolymer is fluorinated ethylene propylene.

10. The cell culture bag according to any one of claims 1 to 9, wherein the first fluoropolymer is densified expanded polytetrafluoroethylene and the second fluoropolymer is fluorinated ethylene propylene.

11. a tube extending from a first end to a second end and defining a cell culture compartment; a first joint at the first end; a second composite film configured to fold over the first joint to form a first lap seam over the first joint; a second lap seam extending longitudinally from the first end to the second end; and A cell culture bag comprising: the tube is formed from a first composite film comprising a first fluoropolymer and a second fluoropolymer; the first fluoropolymer has a first thickness; the second fluoropolymer at least partially penetrates a first thickness of the first fluoropolymer; the first composite film has a second thickness of 0.01 mm to 0.059 mm; The first composite film is O ２ Flux is 2,000 cm ３ / m ２ / atm / day ~ 20,000cm ３ / m ２ / atm / day, the first composite film has a tensile strength of 10,000 psi to 92,000 psi; the first composite film has a total transmittance of at least 70%; and A cell culture bag, wherein a second lap seam is formed by overlapping edges of the first composite film.

12. a second joint at the second end, and The cell culture bag of claim 11 , further comprising a third composite film configured to fold over the second joint to form a third lap seam.

13. 13. The cell culture bag according to claim 11 or 12, wherein the second fluoropolymer penetrates 0.00001 mm to 0.005 mm of the first thickness of the first fluoropolymer.

14. The cell culture bag according to any one of claims 11 to 13, wherein the first composite film has a thickness of 0.02 mm to 0.059 mm.

15. The first composite film has a total organic carbon (TOC) of 1 mg / cm ２ The cell culture bag according to any one of claims 11 to 14, wherein the cell culture bag has a viscosity of less than 1000 MPa.

16. The first composite film is ２ Flux is 8,000 cm ３ / m ２ / atm / day ~ 15,000cm ３ / m ２ The cell culture bag according to any one of claims 11 to 15, wherein the cell culture medium has a flow rate of 1 / atm / day.

17. The cell culture bag according to any one of claims 11 to 16, wherein the first composite film has a tensile strength in at least one direction of 20,000 psi to 92,000 psi.

18. The cell culture bag of any one of claims 11 to 17, wherein the first fluoropolymer is expanded polytetrafluoroethylene and the second fluoropolymer is fluorinated ethylene propylene.

19. The cell culture bag of any one of claims 11 to 18, wherein the first fluoropolymer is densified expanded polytetrafluoroethylene and the second fluoropolymer is fluorinated ethylene-propylene.

20. The cell culture bag of any one of claims 11 to 19, further comprising at least one access port in fluid communication with the cell culture compartment.

21. 21. The cell culture bag of claim 11, wherein the cell culture comprises at least one of connective tissue cells, skeletal cells, cardiac cells, epithelial cells, neural cells, endocrine cells, immune cells, lymphocytes, melanocytes, tumor cells, or a combination thereof.

22. The cell culture bag according to any one of claims 11 to 21, wherein the first composite film and the second composite film are the same.

23. The cell culture bag according to any one of claims 12 or 20 to 22, wherein the third composite film is the same as at least one of the first composite film or the second composite film.

24. The cell culture bag according to any one of claims 11 to 23, wherein the peel strength of the first composite film is 0.25 N / mm to 10 N / mm.

25. 1. A cell culture assembly comprising a plurality of cell culture bags connected in series, Each of the cell culture bags comprises: a body formed from a composite film comprising a first fluoropolymer and a second fluoropolymer, the body defining a cell culture compartment configured to hold a cell culture; the first fluoropolymer has a first thickness; the second fluoropolymer at least partially penetrates a first thickness of the first fluoropolymer; the composite film has a second thickness of 0.01 mm to 0.059 mm; the composite film has a unidirectional tensile strength of 10,000 psi to 92,000 psi; O of the composite film ２ Flux is 2,000 cm ３ / m ２ / atm / day ~ 20,000cm ３ / m ２ / atm / day, The cell culture assembly, wherein the composite film has a total transmittance of 70% to 100%.

26. A first cell culture bag of the plurality of cell culture bags includes a first O ２ a second cell culture bag of the plurality of cell culture bags having a second O ２ having a flux, and The first O ２ Flux and the second O ２ 26. The cell culture assembly of claim 25, which is different from flux.

27. a first cell culture bag of the plurality of cell culture bags having a first volume, a second cell culture bag of the plurality of cell culture bags having a second volume, and 27. The cell culture assembly of claim 25 or 26, wherein the first volume is different from the second volume.

28. a first cell culture bag of the plurality of cell culture bags configured for at least one of cell transfection or cell activation; and The cell culture assembly of any one of claims 25 to 27, wherein a second cell culture bag of the plurality of cell culture bags is configured for cell expansion.

29. a cell culture compartment configured to receive a cell culture; and a composite film connected to the cell culture compartment, the composite film comprising a first fluoropolymer and a second fluoropolymer; A cell culture vessel comprising: the first fluoropolymer has a first thickness; the second fluoropolymer at least partially penetrates a first thickness of the first fluoropolymer; the composite film has a second thickness of 0.01 mm to 0.059 mm; the composite film has a unidirectional tensile strength of 10,000 psi to 92,000 psi; The composite film is O ２ Flux is 2,000 cm ３ / m ２ / atm / day ~ 20,000cm ３ / m ２ / atm / day, The cell culture vessel, wherein the composite film has a total transmittance of 70% to 100%.

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