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Preparation method for serum-free and suspension culture of HEK-293 cells and application of serum-free and suspension culture of HEK-293 cells

A technology of HEK-293 and serum-free medium, which is applied in the field of preparation of HEK-293 cell serum-free and suspension culture, which can solve the problems of contamination of exogenous microorganisms and pathogenic factors, difficulty in large-scale production, and difficult quality control , to achieve high transfection efficiency and toxin production capacity, stable cell growth state, and high repeatability

Active Publication Date: 2021-11-30
IMMUNE CELL BIOTECH CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For single-layer adherent culture of 293 cells, a certain concentration of fetal bovine serum (FBS) is generally used. FBS is not only expensive, but also has the risk of contaminating exogenous microorganisms and pathogenic factors, making it difficult to control the quality, and the output is limited, making it difficult to achieve scale. chemical production

Method used

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  • Preparation method for serum-free and suspension culture of HEK-293 cells and application of serum-free and suspension culture of HEK-293 cells
  • Preparation method for serum-free and suspension culture of HEK-293 cells and application of serum-free and suspension culture of HEK-293 cells
  • Preparation method for serum-free and suspension culture of HEK-293 cells and application of serum-free and suspension culture of HEK-293 cells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1 Preparation of serum-free and suspension culture of HEK-293 cells

[0072] 1. Preparation method:

[0073] (1). Serum-free acclimatization:

[0074] a. Recovery of HEK-293 cells

[0075] Take a frozen HEK-293 cell, put it into a 37°C water bath to thaw, shake

[0076] Let it melt as soon as possible in 45ml DMEM solution containing 10% FBS; mix well, centrifuge at 300g for 5min, discard the supernatant; according to 1.0~1.1×10 7 / dish, the resulting pellet was resuspended with 10ml DMEM medium containing 10% FBS, mixed evenly and cultured;

[0077] b. Culture HEK-293 cells in DMEM medium with decreasing concentration gradient of fetal bovine serum

[0078] When the cell confluency is about 80%, the supernatant is discarded, digested with Tryple and passaged to the cell culture dish. At this time, the medium was replaced with DMEM medium containing 5% FBS, cultured at 37° C. in a 5% CO2 incubator, and the growth status of the cells was observed. Fresh medi...

Embodiment 2

[0102] Example 2: Cellular virus packaging and toxin production capacity

[0103] 1. Method steps:

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Abstract

The invention provides a preparation method for serum-free and suspension culture of HEK-293 cells and an application of the serum-free and suspension culture of the HEK-293 cells, and relates to the preparation method for the serum-free and suspension culture of the HEK-293 cells, a cell line obtained by the preparation method and the application of the cell line. The preparation method is few in passage times and high in domestication efficiency, and the obtained cells are high in viability, good in dispersity and stable in growth state and can meet the requirements of commercialized and large-scale production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method for serum-free and suspension culture of HEK-293 cells, a cell line obtained by the preparation method and an application thereof. Background technique [0002] HEK 293 cell line, also known as hek293 and 293 cells, is a human embryonic kidney 293 cell produced in 1973. As the name suggests, they are a typical cell line derived from human embryonic kidney cells. They are a cell line derived from human embryonic kidney cells. They have the characteristics of high transfection efficiency and easy culture. They are a very commonly used exogenous expression study. Genetic cell lines. [0003] The HEK 293 cell line can be used in a variety of research. For example, it can be used to study drug effects on sodium channels, identifiable RNA interference systems, and nuclear export signals in proteins, among others. More specifically, HEK 293 cells were use...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/073C12R1/91
CPCC12N5/0603C12N5/0686C12N2500/90
Inventor 孙秀莲
Owner IMMUNE CELL BIOTECH CO LTD
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