How do you transfect CHO cells?

The optimized protocol for a 24-well plate to transfect CHO cells is here:

1. Plate 10,000-15,000 CHO cells per well in 0.5 ml of complete growth medium 12-24 hours prior to transfection.
2. Wash with 1xPBS and add 0.5 ml of fresh growth medium.

What does PEI do for transfection?

DNA can be introduced into a host cell by transfection with polyethylenimine (PEI), a stable cationic polymer (Boussif et al., 1995). PEI condenses DNA into positively charged particles that bind to anionic cell surfaces.

What are CHO cells and what are they used for?

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 you make PEI for transfection?

Preparation of PEI Stocks

1. Dissolve 100 mg in 100 mL sterile ddH2O.
2. Stir while slowly adding HCl to pH 7.0.
3. Mix for 10 minutes and then recheck pH.
4. Filter sterilize through 0.22 um filter.
5. Aliquot 500 uL to 1000 uL and store in -80C.
6. Thawed solution can be stored at 4C for up to 2 months, label a tube when thawed.

Why are CHO used?

Many events occurred to bring CHO cells to the forefront in biotechnology. CHO cells became popular because they have a low chromosome number and were used to study genetics. These characteristics have made CHO cells the most important cell line for the production of therapeutic proteins including humanized antibodies.

Are CHO cells transformed?

CHO tk- cells were transformed with the clones gene for herpes simplex virus thymidine kinase (HSV-tk) and were found to have a 50-fold lower frequency of transformation than mouse Ltk- cells at the same DNA dosage.

What is stable cell transfection?

Stable transfection refers either to the permanent expression of the gene of interest through the integration of the transfected DNA into the nuclear genome, or the maintenance of a transfected plasmid as an extra chromosomal replicating episome within the cell.

What cells can be transfected?

Transfection terminology

• Transfection. Transfection commonly refers to the introduction of nucleic acids into eukaryotic cells, or more specifically, into animal cells.
• Transformation. Transformation is often used to describe non-viral DNA transfer in bacteria, non‑animal eukaryotic cells, and plant cells.
• Transduction.

What is PEI coating?

PEI (Polyetherimide), also known as Ultem, is a reusable “relatively maintenance free” build surface for both ABS (with a Heated Bed) and PLA (hot or cold) requiring no additional adhesives such as glue or tape. PEI is appealing due to its “no-surface-prep” conditions, which make it convenient to work with.