The evolution of CHO cells’

role in cell line development

Since the first approval of recombinant insulin and human growth hormone in the early 1980s, a multitude of recombinant protein
therapeutics have been approved by regulatory agencies, notably the FDA in the US and EMA in Europe. Given this significant increase
in successful introduction of biological therapeutic agents, there is a crucial need in the drug discovery space to support more efficient
manufacturing processes that require highly productive cell lines. Consequently, Chinese Hamster Ovary (CHO) cells have emerged as
the gold standard for the manufacturing and regulatory approval of therapeutic proteins.

Why CHO? CHO-ori

1956 CHO-K1
CHO cell line is established in the
Several key properties of CHO cells have
laboratory of Theodore Puck at the Eleanor
driven their establishment as the preferred Roosevelt Institute for Cancer Research.
Kao, Puck and co-workers
cloned the CHO-ori cells and
host cell line for regulatory approvals of CHO-ori cells actively divide and do not
distributed to collaborators.
recombinant therapeutic products: exhibit the limitations on doubling times
Mutational analysis of CHO-K1
(Hayflick Limit) observed in primary cells.
suggests that this cell line is
1968
1 missing a chromosome which
carries a gene necessary
for glycine biosynthesis,
Adaptable to growing in paving the way for selection
suspension culture, which methods utilizing chemicals
is ideal for large scale
CHO-S components in media.

1971
production in bioreactors. Adapted for growth in suspension liquid
culture, this cell line is ideal for scale up
and growth in large-scale bioreactors.
CHO-DXB11

2
Urlaub and Chasin at Columbia University
generated CHO lacking DHFR activity in one
locus and a missence mutation in the other.
DHFR-deficient strains cannot grow unless transfected
Adaptable to growth in
serum-free and chemically
MTX-Induced 1980 with a functional copy of DHFR or in media supplemented
with thymidine. Thus, transfecting DXB11 cells with a
defined (animal-free) media Gene Amplification functional DHFR gene attached to a gene of interest
supplements, which ensures An approach was proposed to (GOI) enable selection of cells only carrying their
reproducibility between amplify genes with the help of an GOI by growing them in thymidine-free media.
different batches of cell culture. antagonist of DHFR, the chemical
component methotrexate (MTX).
1981
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Selection of recombinant cell lines using CHO-DG44
stepwise increases in the MTX concentration Urlaub and Chasin constructed a
in the culture medium resulted in amplified CHO line containing full deletion
copies of the transfected DHFR gene together of the two DHFR loci.
Allow posttranslational
with the GOI. Such induced gene amplification
modifications DXB11 cells were limited in their
1983
usually increased the productivity of the GOI.
(e.g. glycosylations) to usefulness because they could
recombinant proteins spontaneously revert back to a
which are compatible and Activase is ®
functional DHFR enzyme, making
bioactive in humans. approved by FDA selection impossible. CHO-DG44
Human tissue plasminogen activator, eliminated this problem by completely

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deleting the DHFR locus, making
1986
marketed as Activase® (Genentech)
becomes the first therapeutic protein selection for the GOI always possible.
from recombinant mammalian Due to this, CHO-DG44 are amongst
cells to obtain market approval. the most widely used CHO cells
Several chemical selection for industrial protein production.
and gene amplification
systems have been CHO-K1SV
developed for CHO cells,
optimized for higher yield of
Vectors for the Glutamine
Synthetase (GS) system,
1989 Today…
More than 100 new recombinant protein
recombinant protein per cell. first reported in 1987, were therapeutics have been approved
adapted to the CHO cell line. by the US FDA or the EMA.
With the increased interest in viral diagnostics and
treatments, CHO cells are again demonstrating
2000s their versatility. They are being engineered to
express coronavirus antigens in the quest to
Cell line development develop new vaccines, and they are being used
to produce monoclonal antibody therapeutics
can be time-consuming. that may provide new treatments for COVID-19.
Accelerate the discovery with our products:

CHO Growth A ClonePix 2 System CloneSelect Imager

Liquid and semi-solid Automated clone screening and Imaging system to
culture media specifically picking system track track and confirm
for CHO cells monoclonality

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FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
4/21