Historically, cancer cell lines have been cultured almost exclusively in culture media supplemented with significant amounts of fetal bovine serum (FBS). Like other undefined media supplements, fetal bovine serum has unwanted physiological, genetic and epigenetic cellular effects1-4 and is known to cause experimental variability and distort experimental results, e.g. in drug screenings and hormone-related studies.5 For cost reasons, immortal cancer cell lines are widely used in research instead of primary cells. They are an accepted and well-defined model system that serves as a constant source of cells while avoiding the limitations posed by the finite lifespan of normal primary cells 6-9 As long as the limitations inherent in substituting cell lines for primary cells are taken into account, tumor cells. can be used in in vitro models to reflect certain aspects and functionalities of terminally differentiated primary human cells.7 However, poorly defined culture media components such as fetal calf serum are a well-known and significant source of variability.1-5 Thus, a more controlled culture environment is key for obtaining more accurate results with cancer cell line models that will facilitate more accurate data analysis and interpretation.4
The PromoCell Cancer Cell Line Medium XF (C-28077) was designed as a universal serum-free/xeno-free media for culturing most of the commonly used human cancer cell lines. The formulation contains no animal derived components such as fetal calf serum, extracts or hydrolysates and exhibits extremely low lot-to-lot variability with high functionality. Being broadly usable across most common adherently growing cancer cell lines, this new culture media is a cost-effective solution for ensuring efficient and robust cell growth across numerous cancer cell lines.
This protocol describes how cancer cell lines can be switched to the Cancer Cell Line Medium XF for the first time.
Figure 1. Accelerated growth kinetics of HT1080 fibrosarcoma cell lines in Promocell’s Cancer Cell Line Medium XF compared to conventional FBS containing culture conditions.HT1080 cells were plated with 5,000 cells/cm² in Cancer Cell Line Medium XF on fibronectin-coated vessels (red) or in DMEM + 2 mM L-Glutamine + 10% FCS (grey). Subsequently, the cells were cultured for 10 consecutive passages with a passage interval of 3 – 4 days.
Figure 2. Morphology of HT1080 fibrosarcoma cells cultured in Cancer Cell Line Medium XF compared toconventional FBS containing culture conditions.HT1080 cells on day three after subculture (P7) are shown in the Cancer Cell Line Medium XF (B) and conventional FBS containing culture conditions (C) (100x magnification).
Figure 3. Morphology of MCF-7 breast carcinoma cells in the Cancer Cell Line Medium XF.MCF-7 cells were plated at 10,000 cells/cm2 in fibronectin-coated vessels and cultured for three passages in the Cancer Cell Line Medium XF. The cells exhibited efficient proliferation as well as a typical but slightly more compact compared to traditional culture media (not shown). Images were taken 1 day (A) and 5 days (B) after seeding at 100x magnification.