Cell culture vessel and method of production thereof

• US 9,217,130 B2
• Filed: 12/11/2008
• Issued: 12/22/2015
• Est. Priority Date: 12/14/2007
• Status: Active Grant

• Current Assignee: Meijo Nano Carbon Co., Ltd.
• Original Assignee: Meijo Nano Carbon Co., Ltd.
• Inventors: Hashimoto, Takeshi, Akasaka, Tsukasa, Yokoyama, Atsuro, Watari, Fumio
• Primary Examiner(s): GALISTEO GONZALEZ, ANTONIO
• Application Number:
  US12/747,826
• Publication Number:
  US 20100273263A1
• Time in Patent Office: 2,567 Days
• Field of Search: 435/402, 977/712, 977/892, 977/923
• US Class Current: 1/1
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 23/20 Material Coatings immunocoatings C12M25/00
• CPC Class Codes: C12M 35/08 Chemical, biochemical or biological means, e.g. plasma jet, co-culture

Abstract

Disclosed is a method of producing a cell culture vessel (10) having a carbon nanotube (CNT) layer (14) on its surface. The method comprises the steps of providing a vessel (12) having a predetermined shape; providing a CNT dispersion of a CNT material composed primarily of CNT dispersed in a dispersion medium at a concentration of not more than 50 mg/L; and forming the carbon nanotube layer (14) on the surface of the vessel (12). The formation of the CNT layer (14) is achieved by alternately repeating a supply step of applying the CNT dispersion solution to the vessel (12) and a drying step of drying the applied dispersion solution one or more times.

Claims

1. A method of culturing cells comprising:

(a) disposing carbon nanotubes in a dispersion medium to form a mixture thereof, wherein the dispersion medium is a solvent primarily comprised of one of water and a lower alcohol having 1-4 carbons and wherein the carbon nanotubes have a concentration in the mixture of less than or equal to 20 mg/L,

(b) subjecting the mixture to ultrasonic vibrations to form a dispersion, in which the carbon nanotubes are in a highly-dispersed state with no visible agglomerates,

(c) applying an amount of the dispersion in the highly-dispersed state directly to a surface of a vessel such that a resulting carbon nanotube thin-film after drying will contain carbon nanotube material in an amount of at least 0.001 μ

g and not more than 0.1 μ

g per cm²,

(d) drying the applied dispersion on the surface to form the carbon nanotube thin-film,

(e) repeating steps (c) and (d) at least 10 times to form a built-up carbon nanotube layer structure having at least 10 layers of carbon nanotube thin-films and such that the built-up carbon nanotube layer structure contains the carbon nanotube material in an amount of at least 0.02 μ

g and not more than 10 μ

g per cm²,

(f) sterilizing the vessel having the built-up carbon nanotube layer structure,

(g) disposing cells in the sterilized vessel so that the cells directly contact the built-up carbon nanotube layer structure, and

(h) subjecting the vessel having the cells disposed therein to cell culturing conditions;

wherein steps (c) and (d) are carried out in an environment having a relative humidity of less than or equal to 30% and the dispersion contains substantially no polymer ingredients.

2. A method of culturing cells comprising:

(a) disposing carbon nanotubes in a dispersion medium to form a mixture thereof, wherein the dispersion medium is a solvent primarily comprised of one of water and a lower alcohol having 1-4 carbons and wherein the carbon nanotubes have a concentration in the mixture of less than or equal to 20 mg/L,

(b) subjecting the mixture to ultrasonic vibrations to form a dispersion, in which the carbon nanotubes are in a highly-dispersed state with no visible agglomerates,

(c) applying an amount of the dispersion in the highly-dispersed state directly to a surface of a vessel such that a resulting carbon nanotube thin-film after drying will contain carbon nanotube material in an amount of at least 0.001 μ

g and not more than 0.1 μ

g per cm²,

(d) drying the applied dispersion on the surface to form the carbon nanotube thin-film,

(e) repeating steps (c) and (d) at least 10 times to form a built-up carbon nanotube layer structure having at least 10 layers of carbon nanotube thin-films and such that the built-up carbon nanotube layer structure contains the carbon nanotube material in an amount of at least 0.02 μ

g and not more than 10 μ

g per cm²,

(f) sterilizing the vessel having the built-up carbon nanotube layer structure,

(g) disposing cells in the sterilized vessel so that the cells directly contact the built-up carbon nanotube layer structure, and

(h) subjecting the vessel having the cells disposed therein to cell culturing conditions;

wherein steps (c) and (d) are carried out at room temperature and atmospheric pressure and the dispersion contains substantially no polymer ingredients.

3. A method of culturing cells comprising:

(a) disposing carbon nanotubes in a dispersion medium to form a mixture thereof, wherein the dispersion medium is a solvent primarily comprised of one of water and a lower alcohol having 1-4 carbons and wherein the carbon nanotubes have a concentration in the mixture of less than or equal to 20 mg/L,

(b) subjecting the mixture to ultrasonic vibrations to form a dispersion, in which the carbon nanotubes are in a highly-dispersed state with no visible agglomerates,

(c) applying an amount of the dispersion in the highly-dispersed state directly to a surface of a vessel such that a resulting carbon nanotube thin-film after drying will contain carbon nanotube material in an amount of at least 0.001 μ

g and not more than 0.1 μ

g per cm²,

(d) drying the applied dispersion on the surface to form the carbon nanotube thin-film,

(e) repeating steps (c) and (d) at least 10 times to form a built-up carbon nanotube layer structure having at least 10 layers of carbon nanotube thin-films and such that the built-up carbon nanotube layer structure contains the carbon nanotube material in an amount of at least 0.02 μ

g and not more than 10 μ

g per cm²,

(f) sterilizing the vessel having the built-up carbon nanotube layer structure,

(g) disposing cells in the sterilized vessel so that the cells directly contact the built-up carbon nanotube layer structure, and

(h) subjecting the vessel having the cells disposed therein to cell culturing conditions;

wherein steps (c) and (d) are repeated less than or equal to 200 times or between 20-50 times,

wherein the dispersion comprises volatile and non-volatile ingredients and the carbon nanotubes account for at least 95 wt % of the non-volatile ingredients in the dispersion,

wherein steps (c) and (d) are carried out in an environment having a relative humidity of less than or equal to 30% and at room temperature and atmospheric pressure; and

the dispersion contains substantially no polymer ingredients.