Almost every type of cell culture vessel, together with support consumables such as tubes and pipettes, are commercially available as single use, sterile packs. Numerous commercial sources of tissue culture polystyrene (TCPS) vessels are available, creating a wide variety of manufacturers, surface chemistries and culture formats. They are usually treated to provide a hydrophilic surface to facilitate attachment of extensive cell lines. Here, we explore the variations in cell growth performance among such culture vessels, through the corresponding results on cell viability, confluency, and total cell count.
Anecdotally, many researchers have found that cell behaviour depends strongly on the choice of TCPS source. The aim of this study was not to identify or claim that one TCPS vessel source is ‘‘better’’ than others, but rather to explore on the cell growth performance varied and correlated with different TCPS manufacturers.
In this study, we considered numerous samples of commonly used TCPS culture vessels from different manufacturers including Sorfa, Corning, Nunc, and SPD Biomedia. We then investigated whether these differences affected cell behaviour and growth performance. ECC-1 (Human endometrial carcinoma) and MDA-MB-231 (Human breast adenocarcinoma) cells were cultured in DMEM, High glucose medium supplemented with 10% fetal bovine serum (FBS), using TCPS: T25 flask, T75 flask and 6-well plate. All cells were maintained at 37oC in 5% CO2.
Characterization of cell growth performance on TCPS cultureware revealed significant differences among commercial sources. We assessed the cell viability performance through standard MTT assay method. The bar graph shows the cell viability of ECC-1 and MDA-MB-231 cell lines from four different brands of 6-well culture plate. In theory, the greater number of viable and metabolically active cells, the higher the absorbance value.
Interestingly, no statistically significant difference in the cell confluency of both ECC-1 and MDA-MB-231 in both type of vessels tested.
To further evaluate the cell growth performance, both cell lines were seeded in flask and plate, and incubated for 48 hours in complete culture media at 37oC and 5% CO2. Then, cells were prepared for cell counting using Trypan blue solution.
Total cells per ml of ECC-1 and MDA-MD-231 cell lines in four brands of T25 and T75 cell culture flask.
Total cells per ml of ECC-1 and MDA-MD-231 cell lines in four brands of 6-well culture plate.
ECC-1 cell line
MDA-MB-231 cell line
Cell images of ECC-1 and MDA-MB-231 cell lines in TCPS flask after 72 hours captured at 10x magnification.S
It is important to note that we do not intend to conclude (nor can we conclude) that one particular brand of TCPS is superior to another. Results for other cell types and for specific features of cell responses may differ greatly than results presented here. Additionally, the desired response of cell growth performance and interactions can be unique to a given experimental purpose. In summary, it is important to recognize both the significant and insignificant differences that exist across seemingly similar types of TCPS, which may in turn play significant roles in cueing cell growth performance.
We gratefully acknowledge support from the Cytus Sdn Bhd, University of Malaya for research resources and technology.
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