Hi,
I am trying to grow JIMT-1 (breast cancer) cell line. However, the cells fail to attach to the surface of the T25 flask. I have contacted the seller but they only provided me with a generic troubleshooting procedure, which I have been following. I want to try and recover this cell line. So, other than growth supplements, is there any suggestion to work on the same?
Hello Ankita
Did you consider environmental stress? Stress can also prevent attachment of adherent cell to the substratum.
The factors that may cause stress include:
1. Contamination
2. Incubator temperature fluctuations
3. Insufficient or inappropriate cell culture surface and
4.Inappropriate environmental gas mixture
Besides the above factors, you should also consider the growth media as an important factor. For instance, citrate may reduce the availability of calcium to the cell and result in reduced cell attachment. In the growth media without transferrin, citrate competes with ions that form insoluble salts with iron. Keeping more iron in solution increases the free radical activity at the cell surface and causes damage to cell attachment sites. In such a case eliminating or reducing the level of citrate in growth media will be required.
Another possibility could be EDTA contamination in the cell culture media. EDTA is a calcium chelator. Changes in calcium ion concentration are highly likely to affect cell attachment and signaling. Calcium concentration reduction in the medium inhibits cell attachment. In cell growth media without transferrin, EDTA competes with ions that form insoluble salts with iron. By keeping more iron in solution, EDTA increases the chances that iron can catalyze free radical events at the cell surface or in the extracellular matrices, leading to reduced cell attachment. So, if you are using trypsin/EDTA for detachment of cells, wash the cells thoroughly to get rid of EDTA.
Hope this helps!
Best.
Hi Malcom,
This is such a good input, I will check up on it. Thank you for your response.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Enzymology
- Enzymes