Polymeric cell culturing surface having high cell adhesion

Patent metadata

Field	Value
Publication number	US-2022243021-A1
Application number	US-201817268884-A
Country	US
Kind code	A1
Filing date	Dec 7, 2018
Priority date	Aug 13, 2018
Publication date	Aug 4, 2022
Grant date	—

Abstract

A polymeric substrate has a plasma treated contact surface which is roughened, hydrophilic and has a higher oxygen atomic content than the interior portion. The treated contact surface has an enhanced cell adhesion, cell growth and cell recovery rate. During the treatment, the contact surface is contacted with a process gas introduced through a gas inlet near the treated surface and radio frequency electrical power is introduced in the process gas, forming a treated contact surface that has improved cell recovery compared to an untreated contact surface. The process gas optionally can be nitrogen gas, oxygen gas, or a gas that contains nitrogen atoms, oxygen atoms, or a combination of nitrogen and oxygen atoms. The process optionally improves cell recovery of a chicken embryo cell culture from the treated contact surface.

First claim

1 .- 29 . (canceled)

30 . A polymeric substrate comprising a vessel having a wall having an inner surface enclosing a lumen, an outer surface, and an interior portion between the inner surface and the outer surface, and
a treated contact surface, wherein the contact surface comprises at least a portion of the inner surface of the vessel, in which the contact surface has a roughness quantified by at least one of the four parameters below:
a surface area difference A, greater than 0.055%, optionally from 0.06% to 2%, optionally from 0.1% to 1.5%, optionally from 0.5% to 1.2%, optionally from 0.9% to 1.1%;
a root mean square surface slope, Sdq, greater than 1.9°, optionally from 2° to 20°, optionally from 4° to 15°, optionally from 6° to 12°, optionally from 7° to 10°, optionally from 7° to 9°;
a density of summits, Sds, greater than 44.4/μm 2 , optionally from 45/μm 2 to 200/μm 2 , optionally from 50/μm 2 to 180/μm 2 , optionally from 60/μm 2 to 170/μm 2 , optionally from 70/μm 2 to 160/μm 2 , optionally from 80/μm 2 to 160/μm 2 , optionally from 90/μm 2 to 150/μm 2 , optionally from 100/μm 2 to 150/μm 2 , optionally from 110/μm 2 to 150/μm 2 , optionally from 120/μm 2 to 150/μm 2 ; or
a mean summit curvature, Ssc, greater than 3.62/μm, optionally from 4/μm to 50/μm, optionally from 6/μm to 45/μm, optionally from 8/μm to 40/μm, optionally from 10/μm to 35/μm 2 , optionally from 12/μm 2 to 30/μm, optionally from 14/μm to 30/μm, optionally from 16/μm to 30/μm, optionally from 18/μm to 30/μm, optionally from 20x/μm to 25/μm;
wherein the parameters A, Sdq, Sds, and Ssc are each determined by measurement of a 5 μm×5 μm image area on the contact surface; and in which the XPS atomic composition of the interior portion of the polymeric substrate comprises a smaller proportion of oxygen and a greater proportion of carbon than the XPS atomic composition of the contact surface, wherein the XPS atomic composition of the contact surface is determined to a depth of 5.8 Angstroms.

31 . The substrate of claim 30 , in which the interior portion and the contact surface consist essentially of the same polymer and;
the XPS atomic composition of the contact surface comprises from 0.1 to 30 atomic %, optionally from 2 to 30 atomic %, optionally from 5 to 20 atomic %, optionally from 10 to 20 atomic %, optionally from 13 to 16 atomic % more oxygen than the XPS atomic composition of the interior portion; the XPS atomic composition of the contact surface comprises from 0.1 to 20 atomic %, optionally from 5 to 15 atomic %, optionally from 9 to 12 atomic %, carbon atoms to which oxygen is grafted; the contact surface comprises from 0.1 to 20 atomic %, optionally from 5 to 15 atomic %, optionally from 9 to 12 atomic %, hydrogen bond acceptor groups; and the grafted oxygen is present in the form of moieties selected from C—O, CO 3 , C═O, or O—C═O, or any combination of two or more of these, as measured by x-ray photoelectron spectroscopy (XPS).

32 . The substrate of claim 30 , in which the contact surface has a thickness of 0 to 20 nm, optionally 0.1 to 10 nm, optionally 0.2 to 1 nm, optionally 0.2 to 0.7 nm, optionally about 0.6 nm; and in which the contact surface is hydrophilic and has higher cell adhesion than an untreated otherwise identical surface or biological coating coated surface; and the interior portion consists essentially of carbon and hydrogen atoms.

33 . The substrate of claim 32 , in which the contact surface has a thickness of less than 0.6 nm and the XPS atomic composition of the interior portion at a depth of 0.6 nm comprises from 1% to 10% oxygen; alternatively
in which the contact surface has a thickness of less than 1.2 nm and the XPS atomic composition of the interior portion of the treated polymeric substrate at a depth of 1.2 nm comprises from 0.5% to 5% oxygen; alternatively in which the contact surface has a thickness of less than 1.7 nm and the XPS atomic composition of the interior portion of the treated polymeric substrate at a depth of 1.7 nm comprises from 0.3% to 3% oxygen; alternatively in which the contact surface has a thickness of less than 2.3 nm and the XPS atomic composition of the interior portion of the treated polymeric substrate at a depth of 2.3 nm comprises from 0.1% to 1% oxygen; alternatively in which the contact surface has a thickness of less than 2.3 nm and the XPS atomic composition of the interior portion of the treated polymeric substrate at a depth of 2.3 nm comprises from 0.1% to 1% oxygen; alternatively in which the XPS atomic composition of the interior portion of the treated polymeric substrate at a depth of 2.9 nm comprises from 0.1% to 1% oxygen.

34 . The substrate of claim 30 , in which the surface contact angle of the contact surface is from 38° to 62°, optionally from 50° to 70°, optionally from 55° to 65°, optionally from 60° to 64°, optionally from 30° to 50°, optionally from 30 to 40°, optionally from 35° to 45°, optionally from 37° to 41°.

35 . The substrate of claim 30 , in which the vessel comprises a flask, a bottle, or a tube.

36 . The substrate of claim 35 , in which the vessel further comprises a closure, optionally a stopper, cap, lid, top, cork or any combination of them.

37 . The substrate of claim 36 , in which the closure is formed by inserting a plastic or elastomer stopper into a cap.

38 . The substrate of claim 35 , in which the inner surface is generally cylindrical.

39 . The substrate of claim 38 , in which the vessel comprises a roller bottle.

40 . The substrate of claim 39 , in which the inner surface defining the treated contact surface has multiple ribs.

41 . The substrate of claim 30 , in which the vessel has a volumetric capacity from 1 mL to 100 L, optionally from 100 mL to 5 L, optionally 1 L, optionally 2 L.

42 . The substrate of claim 30 , in which the treated polymeric substrate comprises thermoplastic material, for example a thermoplastic resin, for example an injection-molded thermoplastic resin; and in which the thermoplastic resin optionally comprises polystyrene.

43 . The substrate of claim 42 , in which the thermoplastic material comprises a hydrocarbon polymer, for example an olefin polymer, polypropylene (PP), polyethylene (PE), cyclic olefin copolymer (COC), cyclic olefin polymer (COP), polymethylpentene, polystyrene, hydrogenated polystyrene, polycyclohexylethylene (PCHE), or combinations of two or more of these, or a heteroatom-substituted hydrocarbon polymer, for example a polyester, polyethylene terephthalate (PET), polyethylene naphthalate, polybutylene terephthalate (PBT), polyvinylidene chloride (PVdC), polyvinyl chloride (PVC), polycarbonate, polylactic acid, epoxy resin, nylon, polyurethane polyacrylonitrile, polyacrylonitrile (PAN), an ionomeric resin, or any combination, composite, blend, or laminate of any two or more of the above materials.

44 . The substrate of claim 30 , in which the contact surface is treated under conditions effective to improve cell recovery of a chicken embryo cell culture from the treated contact surface relative to the initial contact surface;
the treatment comprising contacting the contact surface with a process gas; and introducing radio frequency electrical power in the process gas adjacent to the initial contact surface to generate plasma adjacent to the initial contact surface, thereby forming a treated polymeric substrate having a treated contact surface; and wherein cell recovery of a chicken embryo cell culture is determined by contacting a cell culture with Medium DMEM containing calf serum, placing 10 mL of the cell culture and culture medium i

Assignees

• Sio2 Medical Products Inc

Inventors

• Taha Ahmad
• Maurer Brian
• Wills Matthew

Classifications

• C12M25/06 Primary

  Plates; Walls; Drawers; Multilayer plates · CPC title

• C12M25/02

  Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00) · CPC title

• C08J2300/22

  Thermoplastic resins · CPC title

• C08J2325/06

  Polystyrene · CPC title

• C08J7/123 Primary

  Treatment by wave energy or particle radiation (C08J7/18 takes precedence) · CPC title