Construction of Dual-3 and Dual-29 cell lines that express NDST2 and 3OST-1 has been previously reported [15 (link)]. CHO-S® (Thermo Fisher Scientific Inc. Waltham, MA), Dual-3, and Dual-29 cells were maintained in 125 mL polycarbonate Erlenmeyer flasks (Corning, Corning, NY) containing 25 mL of CD CHO medium (Thermo Fisher Scientific) supplemented with 8 mM GlutaMAX™ (Thermo Fisher Scientific) and 15 mL of hypoxanthine/thymidine solution per 500 mL of medium (HT, Corning Life Sciences, Corning, NY). All cells were seeded at 2 × 105 cells/mL and cultured on orbital shakers agitated at 125 rpm in a humidified 37 °C incubator with 5% CO2. In addition, 1 mg/mL of Geneticin (G418, Thermo Fisher Scientific) and 500 mg/mL of Zeocin™ (Thermo Fisher Scientific) were added to the medium for dual expressing cell lines. Viable cell densities were determined by trypan blue exclusion using a Bio-Rad TC20™ automated cell counter.
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Construction of Dual-Expressing CHO Cell Lines
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Corresponding Organization : SUNY Polytechnic Institute
Other organizations : Albany College of Pharmacy and Health Sciences, Rensselaer Polytechnic Institute, Thermo Fisher Scientific (United States)
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- Cell lines used: CHO-S, Dual-3, and Dual-29
- Viable cell densities
- Maintained in 125 mL polycarbonate Erlenmeyer flasks containing 25 mL of CD CHO medium supplemented with 8 mM GlutaMAX and 15 mL of hypoxanthine/thymidine solution per 500 mL of medium
- Seeded at 2 × 10^5 cells/mL and cultured on orbital shakers agitated at 125 rpm in a humidified 37 °C incubator with 5% CO2
- 1 mg/mL of Geneticin (G418) and 500 mg/mL of Zeocin added to the medium for dual expressing cell lines
- No positive or negative controls were explicitly mentioned in the provided information.
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