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HomeMammalian Cell CultureCorning® U-shaped 150 and 175 cm2 Flasks Demonstrate Same Cell Growth as Traditional Rectangular Design

Corning® U-shaped 150 and 175 cm2 Flasks Demonstrate Same Cell Growth as Traditional Rectangular Design

Audrey B. Bergeron , Ann Rossi

Corning Inc.

Introduction

Corning’s U-shaped 150 and 175 cm2 flasks incorporate improved design features that benefit the researcher without disrupting culture conditions. Rounded shoulders provide better access for rinsing the growth surface or for scraping to collect cells. Additionally, the U-shaped 150 cm2 flask is shorter in length and has a wider diameter opening than its rectangular counterpart, allowing for better access with cell scrapers and serological pipets. The U-shaped 175 cm2 flask has less height in the horizontal position than the traditional rectangular flask, which makes it easier to hold and takes up less vertical space when stacked during incubation. The U-shaped flasks integrate unique designs for improved ergonomics.

Materials and Methods

Each cell growth evaluation was repeated three independent times with n = 9 per vessel shape and size.

Evaluating Cell Growth Under Standard Conditions

Vero cells were seeded into rectangular and U-shaped flasks at 5 x 103 cells/cm2. Cells were cultured for 3 days in Dulbecco’s Modified Eagle’s Medium supplemented with 10% fetal bovine serum. Cells were harvested and ­enumerated using a cell counter.

Evaluating Cell Growth Under Reduced Serum Conditions

HEK-293 cells were seeded into rectangular and U-shaped flasks at 6 x 103 cells/cm2 and cultured in DMEM with 10% FBS supplemented with 1X nonessential amino acids. The day after seeding, the medium was exchanged with reduced-serum medium containing 1% FBS. Cells were cultured for 2 additional days before harvesting and enumeration.

Crystal Violet Staining

To assess cell confluence across each vessel, Vero cells were seeded into 3 rectangular flasks and 3 U-shaped flasks at 3 x 103 cells/cm2 and cultured for 4 days in DMEM supplemented with 10% FBS. Cells were stained with crystal violet dye.

Results and Discussion

Quality and consistency are critical for successful cell culture experiments. For this reason, it is important to evaluate alterations to any given aspect of an experiment. Something as simple as culture vessel shape could have untoward effects on experimental results. Accordingly, we have demonstrated how two commonly used cell lines perform the same regardless of whether they are cultured in traditional rectangular flasks or Corning U-shaped flasks. Figures 1 and 2 demonstrate no statistical difference between the rectangular and U-shaped vessels for cell growth in standard 10% serum and reduced-serum conditions, respectively. We also assessed the flasks for even cell confluence by culturing Vero cells to full confluence and staining with crystal violet dye. Each flask demonstrated even cell confluence across the vessel (Figure 3).

Similar cell growth

Figure 1. Similar cell growth was obtained under standard culture conditions.Vero cells were cultured for 3 days in (A) 150 cm2 and (B) 175 cm2 flasks, then cells were harvested and enumerated. No statistical difference was observed between both vessels. Paired T-test. n = 9.

Similar cell growth was obtained under reduced-serum

Figure 2. Similar cell growth was obtained under reduced-serum conditions.HEK-293 cells were expanded in 1% FBS in (A) 150 cm2 and (B) 175 cm2 flasks, harvested, and enumerated. Paired T-test. n = 9.

Representative images of confluent crystal-violet stained Vero cells

Figure 3. Representative images of confluent crystal-violet stained Vero cells. Vero cells were seeded at 3,000 cells/cm2 and cultured for 4 days prior to staining with crystal violet dye.

Conclusions

  • There was no statistical difference in Vero cell growth between the rectangular and U-shaped vessels. Crystal violet staining also showed that each flask demonstrated even cell confluence.
  • There was no statistical difference in HEK-293 cell growth between the two designs with a reduced-serum concentration.
Materials
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