These are rat dermal fibroblasts that I've been growing for the past 2 days. However, they're not reaching confluence at all, even after I've changed the media. I make sure that the media (DMEM + 10% FBS) is warm and sterile; however I still see they're not growing. A lot of the cells are also dying. Could anyone suggest next steps?
Hello Zuha Yousuf
From the image, I can observe very few cells. What is the seeding density you are using when you subculture rat dermal fibroblast?
The intercellular signaling mechanism is largely determined by the cell population as well as the surrounding environment. The cell seeding density can influence viability, proliferation and differentiation of cells.
Seeding a lower cell density will result in lack of cell-to-cell interaction and communication. As a result, the cells will show slow growth and eventually die.
On the other hand, seeding a high cell density than the optimum will result in cellular stress as there will be overcrowding and lack of space for cells to interact with each other and grow. Moreover, nutrients in the culture medium will be insufficient and there will be accumulation of metabolic waste causing cells to face stressful conditions.
So, use an optimal cell seeding density when you subculture the cells. Generally, for rat dermal fibroblasts you should seed 0.5 to 0.8 million cells in a T-25 flask.
Hope this will help!
Good Luck!
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