Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Mdck-derived cell lines adapted to serum-free culture and suspension culture and method for preparing vaccine virus using the cells

Inactive Publication Date: 2013-07-18
SK CHEM CO LTD
View PDF2 Cites 28 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides an MDCK-derived cell line that can be prepared by serum-free and suspension culture with very low or no tumorigenicity as compared to original MDCK cells. This cell line has superior proliferation potential and can efficiently produce viruses, particularly influenza viruses. It is suitable for use in the production of viral vaccines.

Problems solved by technology

However, such traditional vaccine sources have several problems.
For example, when it is intended to use fertilized chicken eggs for vaccine production, there are difficulties in raising chickens, managing the fertilized eggs depending on vaccine production schedules and purifying ingredients derived from egg proteins.
However, serum is susceptible to contamination with prions and viruses and its commercial products might vary in quality.
Moreover, high-quality fetal calf serum products derived from Australian and New Zealand calves are highly priced, resulting in an increase in production cost.
Since MDCK cell lines exhibit a strong tendency to attach to other surfaces, large-area culture vessels and carriers are required for large-scale culture, incurring considerable costs.
Specifically, investment costs for culture equipment or carriers are vast and a processing step is needed to remove cells attached to carriers, posing the risk of loss of and damage to the cells.
The second approach has the disadvantage in that the procedure is relatively complicated.
Thus, there exists a danger of potential tumorigenicity when original MDCK cell lines are used for vaccine production.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Mdck-derived cell lines adapted to serum-free culture and suspension culture and method for preparing vaccine virus using the cells
  • Mdck-derived cell lines adapted to serum-free culture and suspension culture and method for preparing vaccine virus using the cells
  • Mdck-derived cell lines adapted to serum-free culture and suspension culture and method for preparing vaccine virus using the cells

Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of MDCK-Derived Cell Lines by Serum-Free Culture and Suspension Culture

[0026]CCL-34, an MDCK cell line, was furnished from ATCC. CCL-34 was cultured in an EMEM medium supplemented with 10% serum in a T-25 flask at 37° C. and 5% CO2. After the cells were expanded, they were cultured in a medium consisting of the EMEM medium and a serum-free medium (50%). It was confirmed whether growth of the cells was normal or not during culture. When growth of the cells was confirmed to be normal, the grown cells were cultured in a medium containing a serum-free medium (75%). This procedure was repeated to obtain a cell line adapted to a serum-free medium (100%). EX-CELL MDCK (Sigma), UltraMDCK (Lonza) and VP-SFM (Invitrogen) may be used as the media for serum-free culture.

[0027]The cell line adapted to the serum-free medium was sufficiently expanded in a T-flask. Thereafter, the expanded cell line was adapted to suspension culture with stirring at a rate of 40-80 rpm in a spinner flas...

example 2

Evaluation of Proliferation Potentials of the Cell Lines

[0028]The MDCK-derived cell lines prepared by culture in the serum-free media were cultured under the conditions indicated in Table 1. The proliferation potentials of the MDCK-derived cell lines were evaluated. The MDCK cell line (ATCC CCL-34) as a control was grown in a medium supplemented with 10% serum.

TABLE 1MDCK-derived cell linesControl (ATCC CCL-34)CultureEX-CELL MDCK (Sigma),EMEM (Lonza)mediaUltraMDCK (Lonza),VP-SFM (Invitrogen)AdditivesL-Glutamine (Lonza) L-Glutamine (Lonza) 2% v / v,2% v / vFetal calf serum (Lonza) 10% v / vCulture37° C., 5% CO2, moist37° C., 5% CO2, moistconditionsCultureT-75 flask (15 ml)T-75 flask (15 ml)volume

[0029]The cell concentrations were about 1.0×105 cells / ml at the beginning of culture. When the cell concentrations reached about 1×106 cells / ml or 3-4 days after the culture, the cells were subcultured. The cell concentrations at the beginning of subculture were adjusted to 1×105 cells / ml.

[0030]Ea...

example 3

Evaluation of Proliferation Profiles and Subculture Stability of the Cell Lines

[0031]After the three kinds of cell lines adapted to serum-free culture and suspension culture were continuously cultured in respective spinner flasks, their proliferation profiles and subculture stability were evaluated. The cell concentrations at the beginning of culture were adjusted to about 4×105 cells / ml. About 3-4 days after the culture, the cell concentrations reached about 2×106 cells / ml or more. The culture was conducted under the following conditions. The results are shown in FIG. 1.

[0032]Initial cell concentration: 4×105 cells / ml

[0033]Culture scale: 50 ml spinner flask

[0034]Spinner rotational rate: 60 rpm

[0035]Culture conditions: 37° C., 5% CO2, moist

[0036]Subculture condition: 3-4 days after culture

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Fractionaaaaaaaaaa
Acidityaaaaaaaaaa
Angular velocityaaaaaaaaaa
Login to View More

Abstract

Disclosed is a Madin-Darby canine kidney (MDCK)-derived cell line. The MDCK-derived cell line is derived from MDCK cells deposited under accession number ATCC CCL-34. The MDCK-derived cell line can be prepared by serum-free culture and suspension culture. Preferably, the MDCK-derived cell line has low or no tumorigenicity. The MDCK-derived cell line is preferably selected from MDCK Sky1023, MDCK Sky10234 and MDCK Sky3851. Further disclosed are a culture method for growing the MDCK-derived cells and a method for producing a vaccine virus using the MDCK-derived cells.

Description

CROSS-REFERENCE TO RELATED APPLICATION[0001]This application is a continuation of International Application No. PCT / KR2011 / 006041 filed on Sep. 6, 2011, which claims priority to Korean Patent Application No. 10-2011-0085902 filed on Aug. 26, 2011, which applications are incorporated herein by reference.TECHNICAL FIELD[0002]The present invention relates to a novel MDCK-derived cell line that can be prepared by serum-free culture and suspension culture without the need to be attached to carriers, a method for preparing the MDCK-derived cell line, and a method for producing a vaccine virus using the MDCK-derived cell line.BACKGROUND ART[0003]Fertilized eggs, mouse brains, primary cells and established cells are generally used as sources for the production of vaccines. However, such traditional vaccine sources have several problems. For example, when it is intended to use fertilized chicken eggs for vaccine production, there are difficulties in raising chickens, managing the fertilized ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): A61K39/145
CPCA61K39/145C12N7/00C12N2760/16252C12N2760/16234C12N2760/16152C12N2760/16134A61K39/12A61K2039/5254C12N5/0686C12N2500/90A61P31/12A61P31/16Y02A50/30A61K39/00C12N5/06C12Q1/02A61K2039/525C12N2760/16151C12N2760/16171C12N2760/16251C12N2760/16271
Inventor PARK, YONG WOOKLEE, KUN SELEE, BONG-YONGPARK, MAHNHOONKIM, HUNKIM, YUN-HEELEE, SU-JEEN
Owner SK CHEM CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com