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

Bovine kidney cells capable of suspension culture and suspension culture method and application thereof

A suspension culture, cell suspension technology, applied in the field of cell engineering, can solve the problems of low production efficiency, occupying a lot of space, and low cell yield per unit.

Active Publication Date: 2021-02-19
JINYUBAOLING BIO PHARMA CO LTD
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

MDBK can usually be cultured with Kjeldahl bottles, spinner bottles or even reactor carriers, but the essence has not changed. The cells are all adherent culture techniques. The adherent culture technique has small batches, large batch-to-batch differences, low production efficiency, and labor-intensive Large, takes up a lot of space, low cell unit yield, complex large-scale production process
[0003] Virus culture or vaccine production such as bovine viral diarrhea / mucosal membrane, bovine infectious rhinotracheitis, and pseudorabies is usually amplified by static culture or spinner bottle culture of MDBK adherent cells, but the scale-up of the adherent culture process is complicated, There are many problems such as small batch size, large batch-to-batch variation, low production efficiency, high labor intensity, and large space occupation, low cell unit yield, and complex large-scale production process
In CN104162154A, it is proposed to use suspension culture technology to produce bovine viral diarrhea and bovine infectious rhinotracheitis dual inactivated vaccine. It uses domesticated MDBK cells and has not formed a suspension cell strain that can be stably passed on. Due to the uncertainty of cell suspension domestication Factors, not every time can be successfully replicated, there are great difficulties in production applications

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
  • Bovine kidney cells capable of suspension culture and suspension culture method and application thereof
  • Bovine kidney cells capable of suspension culture and suspension culture method and application thereof
  • Bovine kidney cells capable of suspension culture and suspension culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1, acquisition and preservation of suspension bovine kidney cell line (suspension MDBK cell line)

[0031] The suspension MDBK cell strain in the present invention is a MDBK cell strain capable of suspension culture obtained by screening ordinary adherent MDBK cells through procedures such as low-serum adherent culture, low-serum suspension culture, and serum-free suspension culture.

[0032] The specific screening process is as follows:

[0033] 1. Adherent MDBK Cell Culture

[0034] Usually, the adherent MDBK cells are cultured at 37°C with 10% (volume percentage, V / V) newborn bovine serum (purchased from: Jinyuankang Bioengineering Co., Ltd.) in commercially available DMEM medium (purchased from: GIBCO), and the cells are After the monolayer is overgrown, digest and disperse with 0.25% (V / V) trypsin, dilute and subculture according to the ratio of 1:3 to 1:5, and expand the culture.

[0035] 2. Low serum adherent culture

[0036] Adherent MDBK cells cov...

Embodiment 2

[0059] Embodiment 2, the acquisition of bovine kidney cell MDBK-S CGMCC No.11795 kinds of cell suspensions

[0060] The method for obtaining bovine kidney cell MDBK-S CGMCC No.11795 kinds of cell suspensions comprises the following steps:

[0061] 1) Recovery of bovine kidney cells MDBK-S CGMCC No.11795

[0062] Take out a suspension of frozen bovine kidney cell line MDBK-S CGMCC No.11795 from liquid nitrogen, thaw quickly in a water bath at 37°C-40°C, centrifuge at 1000rpm for 5min, discard the supernatant, and add the bovine kidney cell suspension above Resuspend the cells in the culture medium, transfer them to the Erlenmeyer shaker flask, add special suspension medium for bovine kidney cells, and adjust the cell density to 0.3-0.5×10 6 Cells / mL, placed in a shaker at 37°C, 100rpm for suspension culture.

[0063] 2) Passaging of bovine kidney cells MDBK-S CGMCC No.11795

[0064] Step 1) Sampling and counting after 48-72 hours of cell culture, if the cell density is ≥1.0×...

Embodiment 3

[0065] Example 3. Reactor fed-batch culture of bovine kidney cells MDBK-S CGMCC No.11795

[0066]Fed-batch culture of bovine kidney cells MDBK-S CGMCC No.11795, including the following levels and scale-up culture:

[0067] 1) Fed-batch culture in 5L reactor

[0068] First use bovine kidney cell-specific suspension medium to dilute the seed cell suspension (prepared in Example 2) of bovine kidney cell MDBK-S CGMCC No.11795 to a density of 0.3-0.5×10 6 cells / mL, then transfer 1000-2000mL of diluted cell suspension (according to the reactor conditions) to a 5L reactor, and set the cell culture conditions: temperature 37°C, dissolved oxygen 40-60% (saturation percentage), pH value 7.0-7.2, rotation speed 40-60rpm, until the cell density reaches 0.8×10 6 When cells / mL (judging that the cells are growing vigorously), start feeding the special suspension medium for bovine kidney cells, the flow rate is 1000mL / day, stop feeding when the culture volume reaches 5L, and take it out to ...

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
diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses bovine kidney cells capable of suspension culture and a suspension culture method and application thereof. The bovine kidney cell MDBK suspension culture method of the present invention saves a large number of manual operations such as cell digestion, liquid replacement, frequent sorting, etc., the culture method is simple and fast, and the reactor culture is used to realize process automation, which not only improves the cell yield per unit volume , It also solves the technical problem of differences between batches of traditional adherent cultured cells. The bovine kidney cell MDBK and the suspension culture cell thereof of the present invention can be used in fields such as the cultivation of various sensitive viruses, the production of antibody proteins, the production of vaccines, the inspection of exogenous viruses in products, and the like.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, in particular to the application of suspensible cultured bovine kidney cells (MDBK) in virus culture and vaccine production after suspension culture. Background technique [0002] The bovine kidney cell (MDBK) passage cell line is derived from bull kidney cells, which can be used for virus culture, raw material inspection, and nutrient metabolism research. MDBK can usually be cultured by Kirschner flasks, spinner bottles or even reactor carriers, but the essence has not changed. The cells are all adherent culture processes. The adherent culture process has small batches, large batch-to-batch differences, low production efficiency, and labor intensity. It is large, takes up a lot of space, has a low yield per unit of cells, and has a complex large-scale production process. [0003] For virus culture or vaccine production of bovine viral diarrhea / mucosal membrane, bovine infectious rhinot...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N5/02C12R1/91
CPCA61K39/12A61K2039/5252A61K2039/5254C12N5/0686C12N7/00C12N2710/16034C12N2710/16051C12N2710/16063C12N2710/16734C12N2710/16751C12N2710/16763C12N2710/24234C12N2710/24251C12N2710/24263C12N2760/18534C12N2760/18551C12N2760/18563C12N2770/24334C12N2770/24351C12N2770/24363
Inventor 郝鹏刘国英温谢孔彩平纪燕苍枫商晓桂
Owner JINYUBAOLING BIO PHARMA 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