What are retrovirus like particles?
Each retrovirus particle contains two copies of the viral genomic particle RNA (pRNA) molecule. Therefore, quantification of retrovirus particles can be achieved by quantifying the pRNA copy number, i.e. every two copies of retroviral pRNA is equivalent to one retrovirus-like particle.
What is the doubling time of CHO cells?
The doubling time is typically 24 hours for contact-inhibited adherent CHO-K1 cultures but can decrease to nearly half that for suspension CHO cells.
What do CHO cells produce?
CHO cells are the most common mammalian cell line used for mass production of therapeutic proteins. They can produce recombinant protein on the scale of 3-10 grams per liter of culture.
How do CHO cells grow in suspension?
CHO cells grow quickly and easily and cell doubling time is 14-17 hours. Add 3 mL of Trypsin-EDTA to flask and watch for cell layer detachment under an inverted microscope. This should occur within 5-15 minutes. Do not agitate cells during this type as agitation encourages clustering.
Are CHO cells transformed?
The U-code construct was inserted into the nuclear genome (nucDNA) of a wild type CHO cell. The expressed transgene products enabled the transformed CHO cell lines to grow in up to 1000 ng mL(-1) oligomycin, while untransformed sensitive CHO cells were eliminated in 1 ng mL(-1) oligomycin.
What is the morphology of CHO cells?
Nickel(II) compounds are known human and animal carcinogens. In this study, the effects of nickel(II) acetate on cell cycle, apoptosis and p53 expression were investigated in order to unveil the elements of early cellular responses to the metal.
Are CHO cells stem cells?
Given that Chinese hamster ovary (CHO) cells are the predominant host cells used for therapeutic protein production and no pluripotent stem cell line or other normal cell line has been isolated from Chinese hamster, this iPSC line may serve as a useful tool for research using CHO cells or even be used for deriving new …
How do cells adapt to suspension?
Begin with cultures at maximum cell density. Dissociate the cell monolayer using standard procedures. Centrifuge and resuspend the cell suspension with serum-free medium containing 5% serum (V/V).
How to remove Retrovirus particles from CHO-cell derived products?
Removal of endogenous retrovirus-like particles from CHO-cell derived products using Q sepharose fast flow chromatography Retrovirus-like particles (RVLPs) that are expressed during the production of monoclonal antibodies in Chinese hamster ovary (CHO) cell cultures must be removed during product recovery.
How to quantify Retrovirus particles?
Therefore, quantification of retrovirus particles can be achieved by quantifying the pRNA copy number, i.e. every two copies of retroviral pRNA is equivalent to one retrovirus-like particle.
Do retroviruses that stably Express transgene trigger inflammation?
Retroviruses that stably express transgene exhibit lower potential in triggering inflammation than adenoviruses and herpes viruses that usually produce transient transgene expression but inflammation in the host cell ( Lee et al., 2017 ).
What is the difference between retroviruses and adenoviruses?
Generally, retroviruses can only be used to transfect dividing cells while adenoviruses, AAVs and herpes viruses can be used to transfect both dividing and non-dividing cells ( Lee et al., 2017 ). However, viral transduction is associated with higher cytotoxicity and may pose a risk for viral infection ( Kim & Eberwine, 2010; Mali, 2013 ).