What causes cells to clump?
The most common cause of cell clumping is the presence of free DNA and cell debris in the culture medium, which occurs following cell lysis. The sticky nature of DNA causes cells and other debris to aggregate into large clumps.
How do you stop cell clumping?
Cell clumps can also be reduced through proper handling and equipment usage. If a centrifuge is being used to separate or mix a sample, setting it to the correct speed can reduce the chances of buildup.
Do THP-1 cells adhere?
THP-1 cells are non-adherent, so all treatments you perform to promote their attachment are going to have an impact on them. For instance, PMA tends to upregulate the expression of some genes in differentiated macrophages, which could affect the gene expression induced by other stimuli.
How do THP-1 cells grow?
The THP-1 (ATCC TIB-202) cell culture is usually expanded by the addition of complete culture medium. Full fluid changes should be performed by centrifuging and resuspending the cells in fresh media at least every 7 days.
What does it mean when your blood cells stick together?
In hematology, red cell agglutination or autoagglutination is a phenomenon in which red blood cells clump together, forming aggregates. It is caused by the surface of the red cells being coated with antibodies.
Does Mycoplasma affect cell growth?
In fact, depending on the laboratory, anywhere from 10% to 85% of cell lines may be contaminated. Mycoplasmas can drastically alter your cells and consequently, skew your research results.
What is a single cell suspension?
A cell suspension or suspension culture is a type of cell culture in which single cells or small aggregates of cells are allowed to function and multiply in an agitated growth medium, thus forming a suspension.
How do you remove a THP-1 cell?
“PBS-EDTA at 4 ° C” did not work on my THP-1…. -_-!! replace the Medium with pre warmed PBS and then add 1-2mL of 1x trypsin-EDTA and incubate at 37 for 5 minutes. All cells will be detached.
How do you revive THP-1 cells?
Reviving THP1 cells – (Jul/06/2008 )
- Thaw the frozen stock in warm water by gently swirling.
- In the hood, transfer the contents after pipeting once gently to a 15ml centrifuge tube.
- Add 9 ml of RPMI mediium with 10% FCS, drop wise.
- Spin at 1300 rpm for 10 mins.
- Decant the supernatant and resuspend pellet in fresh medium.
What are THP-1 cells used for?
THP-1 is a human monocytic cell line derived from an acute monocytic leukemia patient. It is used to test leukemia cell lines in immunocytochemical analysis of protein-protein interactions, and immunohistochemistry.
How to break up THP-1 clumps?
How big are the clumps? THP-1 cells grow in clumps as when they divide the new cell attaches to the one it split from. With my THP-1 gentle pipetting up and down will break the clumps but the cells remain viable.
Why do my THP-1 cells grow in clumps?
THP-1 cells grow in clumps as when they divide the new cell attaches to the one it split from. With my THP-1 gentle pipetting up and down will break the clumps but the cells remain viable. If you split the culture often you should have small clumps 2-4 cells, if you leave them for a few days the clumps will grow larger as the cells replicate.
Will THP-1 cells come back from -80°C?
THP-1’s will come back from -80°C. Thaw your cells using the tips above from -80 o C but this time, freeze the cells down using the correct procedure. Transfer them to liquid nitrogen after 24 hours this time! I have witnessed contamination of a lot of things—trypan blue, DMSO, FBS, you name it, and I’ve seen it.
How do you prepare THP1 cells for PCR?
1- Add 180 µl of THP1 cell suspension per well of a 96-well plate (~ 100 000 cells/well). 2- Treat THP1 cells with 20 µl of PMA (final concentration 20 – 50 ng/ml) for 3 hours at 37°C in 5% CO2. 3- Wash cells gently with pre-warmed PBS and add 200 µl supplemented RPMI.