Hello, I have been using MCF 7 cell for 3 months. The newly thawed cell vial was observed to have cells with different morphology. These round shaped cells are also attached to the flask. However with time the population of cells with normal morphology increase. Any kind suggestions on why abnormal morphology in MCF7 flask.
Thank You
Hello Ambulugala Gamage Rajika Greshamali Jinadasa
The cells may be under stress due to which it may be showing such characteristic on thawing. There could be many reasons, some of which are given below.
1. The freezing medium used to freeze MCF-7 cells. We usually use 70% Basal medium + 20% FBS + 10% DMSO.
2. Gradual freezing at -1 degree C/min using Mr. Frosty freezing container must be carried out when mammalian cells are frozen.
3. Quick thawing (< 1 minute) must be performed in a 37°C water bath.
4. Check for mycoplasma as they slowly spread in culture and affect cell morphology and gene expression.
5. For cell revival you may initially use 20% FBS in complete media and maintain the cells in it until they regain their normal morphology.
6. The passage number is an important factor. It is a well-established fact that all cell cultures have the potential to change with time due to selective pressures in the culture environment, and particularly with continuous cell lines, due to genetic instability. Cells may continue to proliferate but as the passage number increases their phenotype and genotype can change.
Best.
Cells in culture are massively rounded in two cases: 1) the population of cells is synchronized and all of them (or almost all) are preparing for mitotic division; 2) cells have trophic problems (an untimely change of the nutrient medium, a jump in temperature or composition of the gas phase in the thermostat, etc.).
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