Culture medium composition, and method for culturing cell or tissue using said composition

A medium and composition technology, applied in general culture methods, animal cells, cell culture medium, etc., can solve the problems of complicated operation of cell culture in culture vessels, reduced cell viability and reproducibility, etc.

Active Publication Date: 2015-05-20
NISSAN CHEM IND LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] However, those hepatocyte culture methods have various problems in that the production method of the culture vessel and the operation of cell culture are complicated, the operations of recovering cells from a support such as collagen and evaluating the function of hepato

Method used

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  • Culture medium composition, and method for culturing cell or tissue using said composition
  • Culture medium composition, and method for culturing cell or tissue using said composition
  • Culture medium composition, and method for culturing cell or tissue using said composition

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0287] (Analysis Example 1: Viscosity measurement and cell suspension test of medium containing deacylated gellan gum)

[0288] Preparation and viscosity measurement of culture medium containing deacylated gellan gum

[0289] Deacylated gellan gum (KELCOGEL CG-LA, manufactured by SANSHO Co., Ltd.) was suspended in pure water to 0.4% (w / v), and dissolved by heating and stirring at 90°C. Allow the aqueous solution to cool to room temperature while stirring, and sterilize in an autoclave at 121°C for 20 min. Place 2 times the concentration of DMEM / F-12 medium (manufactured by Aldrich, 50 mL) and sterile water (47.5 mL) in a 300 mL high beaker, and pass through a homomixer at room temperature (3000 rpm) ) With stirring, the aqueous deacylated gellan gum solution (2.5 mL) was added, and the mixture was continuously stirred for 1 min to prepare a deacylated gellan gum medium composition with a final concentration of 0.01%. A medium composition supplemented with an aqueous deacylated ge...

Example

[0292] (Comparative example) Preparation of medium containing methylcellulose and collagen

[0293] Preparation of medium containing methylcellulose

[0294] DMEM / F-12 medium (manufactured by Aldrich, 100 mL) was placed in a 200 mL pear-shaped bottle, and methylcellulose (M0387, manufactured by Aldrich, 0.1 g) was added. The mixture was stirred while cooling on an ice bath to dissolve the methyl cellulose. Using this solution, a medium composition in which an aqueous methylcellulose solution was added at a final concentration of 0.1, 0.3, 0.6, or 1.0% (w / v) was prepared.

[0295] Preparation of collagen-containing medium

[0296] Add 10-fold concentration of DMEM / F-12 medium (manufactured by Aldrich, 1 mL), buffer for reconstitution (manufactured by Nitta Gelatin Inc., 1 mL), and pure water (1.5 mL) to 0.3% Type IA cell matrix (manufactured by Nitta Gelatin Inc., 6.5 mL), and the mixture was stirred on ice to generate a medium containing 0.2% collagen. Similarly, a medium compositi...

Example

[0303] (Experimental Example 1: Cell proliferation test by scattered single cells)

[0304] Deacylated gellan gum (KELCOGEL CG-LA, manufactured by SANSHO Co., Ltd.) was suspended in ultrapure water (Milli-Q water) to 0.3% (w / v), and stirred by heating at 90°C Come to dissolve. The aqueous solution was sterilized in an autoclave at 121°C for 20 minutes. Use this solution by adding deacylated gellan gum at a final concentration of 0.015% (w / v) to IMDM containing 10% (v / v) fetal bovine serum and 10 ng / mL thrombopoietin (manufactured by WAKO) Medium (manufactured by Gibco) to prepare the medium composition. Immediately afterwards, the human leukemia cell line UT7 / TPO was placed in the medium composition supplemented with the above-mentioned deacylated gellan gum to 20,000 cells / mL, and 5 mL / well was dispersed on a 6-well flat-bottomed microculture plate (from Corning Incorporated). Similarly, the human cervical cancer cell line HeLa was spread at 20000 cells / mL by adding 0.015% (...

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Abstract

The present invention provides a culture method of cells and/or tissues including culturing cells and/or tissues in a suspended state by using a medium composition wherein indeterminate structures are formed in a liquid medium, the structures are uniformly dispersed in the solution and substantially retain the cells and/or tissues without substantially increasing the viscosity of the solution, thus affording an effect of preventing sedimentation thereof, and the like.

Description

technical field [0001] The present invention relates to a medium composition containing a structure capable of suspending cells or tissues, and a method of culturing cells or tissues by using the medium composition. The medium composition and the cell culture method using the medium composition of the present invention can be preferably used for culturing cells and / or tissues of animals or plants, especially in a suspension state. Background technique [0002] In recent years, techniques have been developed for in vitro proliferation or maintenance of various organs, tissues and cells that serve different functions in animal and plant organisms. Proliferating or maintaining organs and tissues in vitro is referred to as organ culture and tissue culture, respectively, and proliferating, differentiating or maintaining cells isolated from organs or tissues in vitro is referred to as cell culture. Cell culture is the technique of proliferating, differentiating or maintaining iso...

Claims

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Application Information

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IPC IPC(8): C12N1/00
CPCC12N5/0025C12N5/0062C12N5/0068C12N2500/50C12N2533/70A01H4/002C12N5/0018C12N5/0693C12N5/0696C12N2500/10C12N2500/30C12N1/00
Inventor 西野泰斗金木达朗大谷彩子猿桥康一郎户村美沙代岩间武久堀川雅人中辻宪夫尾辻智美
Owner NISSAN CHEM IND LTD
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