WAYNE293 LVPRO cell adapted to serum-free culture medium environment and application of WAYNE293 LVPRO cell

Abstract

The invention discloses a WAYNE293LVPRO cell adapted to a serum-free culture medium environment and application of the WAYNE293LVPRO cell. The human embryonic kidney cell WAYNE 293 is preserved in China General Microbiological Culture Collection Center (CGMCC) on May 24, 2021, the address is Institute of Microbiology, Chinese Academy of Sciences, No.3, No.1 Yard, Beichen West Road, Chaoyang District, Beijing, the postal code is 100101, and the preservation number is CGMCC No.22348. The human embryonic kidney cell WAYNE 293 provided by the invention is derived from suspension culture of HEK293 cells in a serum-free culture medium without an anti-caking agent, has the characteristics of short doubling time, high growth density, high virus packaging efficiency and the like, and is an important support for industrial development in the field of cell and gene therapy.

Description

Technical field: [0001] The invention belongs to the field of cells, and in particular relates to a WAYNE293 LVPRO cell adapted to a serum-free culture medium environment, short doubling time, high cell growth density and high virus packaging efficiency and application thereof. Background technique: [0002] With the development of the cell and gene therapy industry, the method of using viral vectors to introduce target genes has become a new direction for human treatment of major diseases such as congenital genetic diseases, leukemia, and cancer. [0003] At present, the preparation of viral vectors is mainly carried out by using HEK293 as host cells and using chemical reagents such as cationic polymers for transient transfection. This is because HEK293 has certain advantages in transient transfection efficiency and virus packaging yield compared with other cells. However, most of the currently marketed gene and cell therapy drugs still use adherent culture, such as the me...

AI Technical Summary

Problems solved by technology

Adherent culture has insurmountable difficulties in the late stage of industrialization, because the number of cells is limited by the surface area of ​​the culture flask, so in the process of large-scale industrial production, it is necessary to expand the production scale of viral vectors, and the investment in equipment and Artificial will greatly increase the cost of virus vector preparation. At the same time, since the adherent culture process still needs to add animal-derived components of fetal bovine serum as a necessary cell culture raw material, this will affect the safety of gene and cell therapy products. brings inevitable risks

[0004] In addition, on the existing serum-free suspension culture HEK293 technology platform, due to the pursuit of subsequent transient transfection efficiency, the use of anti-aggregating agents in the medium is generally reduced or even completely eliminated, and HEK293 cells as a human Embryonic kidney cells are prone to cell clumping during serum-free suspension culture, which greatly affects cell viability, density and transfection efficiency

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