I recently received a fibroblast line XPE that came from primary culture. I inmediately began culturing the cells at DMEM high glucose and pyruvate, 10% FBS Gibco in Corning plates, most of them began to die and seem to have a small growth rate, if any.
A number of factors may be contributing to poor proliferation and survival of the primary human fibroblasts (Is it an immortalized line?). Therefore, as noted by Wassim, adding growth/survival factors may help. However, it would not guarantee healthy proliferation rate. It would depend on optimizing the culture conditions (including the cell density) and using reagents that were used to maintain the primary cultures.
Thanks you for both answers.
Divaker, it is a primary culture.
They are looking better today, after one week with 20% FBS.
When they reached a higher cell density, they improved proliferation rate. It´s probably that they are more dependent of their conditioned media than a inmortalized cell line.
However, I will consider to add growth factors when I ´ll begin to split them
Thanks by advance for your help.
Splitting cultures at 1:2 or 1:3 may help with proliferation rate initially (when cells are adapting to new culture conditions).
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