How are THP-1 cells different?
THP-1 monocytes are differentiated into macrophages by 24 h incubation with 150 nM phorbol 12-myristate 13-acetate (PMA, Sigma, P8139) followed by 24 h incubation in RPMI medium. Macrophages were polarized in M1 macrophages by incubation with 20 ng/ml of IFN-γ (R&D system, #285-IF) and 10 pg/ml of LPS (Sigma, #8630).
How do you transfect a cell line?
Electroporation. Electroporation is a physical method of transfection that involves first suspending your cells and DNA construct in an electroporation buffer. High-voltage pulses of electricity are then applied to the mixture, which creates a potential difference across the cell membrane.
Do THP-1 cells express CD14?
Some studies have reported almost 90% CD14 expression in unstimulated THP-1 cells 28 29 while others have reported lower or no expression 20 30. Similarly, reports describing THP-1 CD14 levels in response to PMA varies from poor or no response 31, 32 to increased levels 33 34 20.
How do I get a cell line?
The simplest way to create a new cell line is to modify an existing one, a common strategy when an established line already comes close to meeting the requirements. Cells optimized to grow particular viruses or maximize recombinant protein production often come from such modifications.
Are THP 1 cells suspension cells?
Normally THP-1 cells grow in suspension, but they can be induced to differentiate into adherent macrophage-like cells, which express IL-8 and other markers, and can thus serve as a model system for differentiation.
What are J774 cells?
Macrophage cell lines like J774 cells are ideal model systems for establishing the biophysical foundations of autonomous deformation and motility of immune cells.
How is the transfection of THP-1 cells efficient?
The protocol maintains cell viability and functionality, thus allowing efficient transfection of THP-1 cells combined with subsequent differentiation of transfected THP-1 cells into mature macrophages. ubiquitin-conjugating enzyme E2D 2.
Is there a non viral transfection of THP-1 monocytes?
Non-viral transfection of THP-1 monocytes has also been described ( Martinet et al., 2003, Lee et al., 2005 ), but without maintaining viability and functionality during differentiation of transfected THP-1 cells into macrophages. Therefore, most studies using transfected THP-1 cells avoid maturation.
Can you use Lipofectamine LTX on THP-1 cells?
Follow these important guidelines when transfecting DNA into THP-1 cells using Lipofectamine® LTX Reagent: The addition of antibiotics to media during transfection may result in cell death, and has not been tested for THP-1 cells. If you wish to use antibiotics during transfection, test your conditions thoroughly.
Which is the best cell line for THP-1?
Many in vitro studies therefore use immortalized proliferating cell lines, such as human THP-1 as well as murine J774A.1 and RAW 264.7, instead.
