Culture medium composition and method of culturing cells or tissues using same
A technology for culturing cells and compositions, applied in general culture methods, animal cells, cell culture media, etc., can solve the problems of complex cell culture operations in culture vessels, reduced cell viability and reproducibility, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0286] (Analytical Example 1: Viscosity Measurement and Cell Suspension Test of Medium Containing Deacylated Gellan Gum)
[0287] Preparation and viscosity measurement of medium containing deacylated gellan gum
[0288] Deacylated gellan gum (KELCOGEL CG-LA, manufactured by SANSHO Co., Ltd.) was suspended in purified water to 0.4% (w / v), and dissolved by heating and stirring at 90°C. The aqueous solution was allowed to cool to room temperature with stirring and sterilized in an autoclave at 121 °C for 20 min. Put 2-fold concentration of DMEM / F-12 medium (manufactured by Aldrich, 50 mL) and sterilized water (47.5 mL) into a 300 mL tall beaker while passing through a homomixer (homomixer) at room temperature (3000 rpm ) was stirred, an 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%. Medium compositions supplemented with aqueou...
experiment Embodiment 1
[0302] (Experimental Example 1: Cell Proliferation Test by Dispersing Single Cells)
[0303] 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 to dissolve. The aqueous solution was sterilized in an autoclave at 121° C. for 20 minutes. Using this solution, deacylated gellan gum was added to IMDM containing 10% (v / v) fetal bovine serum and 10 ng / mL thrombopoietin (manufactured by WAKO) at a final concentration of 0.015% (w / v). 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 20000 cells / mL, and dispersed to a 6-well flat-bottomed microculture plate at 5 mL / well (from Corning Incorporated). Similarly, the human cervical cancer cell line HeLa was plated at 20000 cells / mL by adding 0.015...
experiment Embodiment 2
[0307] (Experimental Example 2: Cell proliferation test by culturing cell line-derived spheres)
[0308] The human liver cancer cell line HepG2 (manufactured by DS PHARMA BIOMEDICAL CO., LTD.) was suspended in DMEM medium containing 10% (v / v) fetal bovine serum (manufactured by WAKO) at 250000 cells / mL, and the The suspension (10 mL) was plated on EZ SPHERE (manufactured by ASAHI GLASS CO., LTD.), and 2 Incubator (5% CO 2 ) for 7 days. Similarly, the human cervical cancer cell line HeLa (manufactured by DS PHARMA BIOMEDICAL CO., LTD.) was suspended at 250000 cells / mL in EMEM medium containing 10% (v / v) fetal bovine serum (manufactured by WAKO) , and the suspension (10 mL) was plated on EZ SPHERE (manufactured by ASAHI GLASS CO., LTD.), and 2 Incubator (5% CO 2 ) for 7 days. A suspension (2.5 mL) of spheres (100 - 200 μm in diameter) obtained here for each cell line was centrifuged (200G, 5 min) to allow the spheres to settle, and the supernatant was removed. Next, the ab...
PUM
| Property | Measurement | Unit |
|---|---|---|
| viscosity | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More