Non-Treated Cell Culture Flasks for Suspension Cells and Insect Cell Culture

Non-treated cell culture flasks are sterile polystyrene T-flasks with an unmodified hydrophobic surface (contact angle ~75–90°) designed for suspension mammalian cells (CHO-S, HEK293-F, Jurkat, K562), hematopoietic progenitors, hybridomas, insect cells (Sf9, Hi5), and microbial culture. NEST Scientific non-treated flasks (T25–T225) are E-beam sterilized to SAL 10⁻⁶, DNase/RNase-free, ISO 9001:2015 and ISO 13485 certified, and available with 0.2 µm vented or plug seal caps.

MBP carries the full non-treated flask line with direct PO ordering for labs across the USA, Canada, and worldwide. For further details, contact customerservice@mbpinc.net

Non-Treated Cell Culture Flasks

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What are Non-Treated Cell Culture Flasks?

Non-treated cell culture flasks are sterile polystyrene T-flasks that retain the original hydrophobic, unmodified surface of the polystyrene substrate — no plasma, corona, or chemical surface treatment is applied. The hydrophobic surface minimizes cell attachment, making these flasks the correct vessel for cell types that grow in suspension or require minimal surface contact: suspension-adapted CHO, Jurkat, K562, HL-60, hematopoietic progenitors, hybridomas for monoclonal antibody production, insect cell lines (Sf9, Hi5), yeast, and bacteria. NEST Scientific non-treated flasks are manufactured from 100% virgin polystyrene in ISO 8 cleanrooms, E-beam sterilized to SAL 10⁻⁶, and available in T25, T75, T175, and T225 sizes with vented (0.2 µm PTFE membrane) or plug seal caps.

Use non-treated flasks exclusively for cell types that do not require surface attachment; placing adherent mammalian cell lines in a non-treated flask causes cell death within 24–48 hours.

What you will find:

Suspension-Optimized Surface, like naturally hydrophobic polystyrene, ensures cells remain in suspension without surface interference.
Scalable Formats precision-engineered in T25, T75, T175, and T225 sizes to fit various research scales.
Aseptic Security with certified RNase, DNase, and Pyrogen-free to maintain the highest purity standards.
Ergonomic Access with angled neck design facilitates seamless pipette access and reduces the risk of contamination during media changes.

How to Choose Non-Treated Cell Culture Flasks

Application: Suspension Mammalian Cells
Suspension-adapted cell lines — CHO-S, HEK293-F, Expi293, and HEK293-EBNA variants used in transient transfection — require non-treated flasks for static suspension culture between spinner flask passages or when the scale is too small for orbital shaker flasks. Cells must be agitated (shaken, pipetted, or rocked) periodically to prevent settling when in static non-treated flasks with volumes >5 mL.

Application: Hematopoietic and Immune Cells
Non-adherent hematopoietic cell lines (Jurkat T-cells, K562 erythroleukemia, HL-60 promyelocytes, Raji B-cells) and primary hematopoietic cells (peripheral blood mononuclear cells (PBMCs), bone marrow progenitors) must be cultured in non-treated vessels. TC-treated surfaces activate integrin signaling in hematopoietic cells, inducing undesired differentiation and stress responses.

Application: Hybridoma Culture
Hybridoma cells for monoclonal antibody (mAb) production are non-adherent B-cell fusions that grow as suspension cultures. Non-treated T75 and T225 flasks, combined with HT-supplemented DMEM/HAT medium, are the standard vessels for hybridoma selection and expansion. Scale-up from T225 non-treated flasks typically progresses to spinner or hollow-fiber bioreactors.

Application: Insect Cells
Sf9 (Spodoptera frugiperda) and Hi5 (Trichoplusia ni) insect cells used in baculovirus expression vector systems (BEVS) require non-treated polystyrene for monolayer culture. On TC-treated surfaces, Sf9 cells attach so strongly that they are difficult to recover. Non-treated flasks allow Sf9 to loosely adhere and detach easily with tapping, or to grow as a loosely attached monolayer that tolerates trypsin-free passaging.

Application: Microbial Culture
Non-treated polystyrene T-flasks with plug seal caps can serve as culture vessels for E. coli, Bacillus subtilis, yeast (S. cerevisiae, P. pastoris), and other microorganisms where the large surface area per volume is needed, and standard Erlenmeyer flask orbital shaking is unavailable. Ventilated caps support aerobic culture; confirm that the vented 0.2 µm membrane is compatible with your aeration requirements.

Cap Selection for Non-Treated Flasks
Vented caps (0.2 µm PTFE membrane) are standard for suspension cultures in a CO₂ incubator. For microbial aerobic culture without a CO₂ incubator, vented caps with 0.2 µm pore still provide adequate aeration. Plug seal caps are used for anaerobic culture, transport, and short-term storage.

Compare Non-Treated Flask SKUs

Non-treated T25, vent cap: Growth Area: 25 cm²; Working Volume: 5–10 mL; NEST Catalog: 707003; Application: Small-scale suspension culture, seed stock.

Non-treated T75, vent cap: Growth Area: 75 cm²; Working Volume: 15–25 mL; NEST Catalog: 708003; Application: Routine suspension maintenance.

Non-treated T175, vent cap: Growth Area: 175 cm²; Working Volume: 35–50 mL; NEST Catalog: 709003; Application: Scale-up, CHO transfection, hybridoma.

Non-treated T225, vent cap: Growth Area: 225 cm²; Working Volume: 50–70 mL; NEST Catalog: 710003; Application: High-yield suspension expansion.

Non-treated T75, plug seal: Growth Area: 75 cm²; Working Volume: 15–25 mL; NEST Catalog: 708004; Application: Anaerobic, transport, sealed storage.

Specifications Context

NEST Scientific non-treated cell culture flasks are manufactured from 100% virgin polystyrene with no surface modification. Contact angle of non-treated PS surface is approximately 75–90° (hydrophobic), confirmed by water contact angle measurement per NEST lot QC data. E-beam sterilized in ISO 8 cleanrooms to SAL 10⁻⁶; DNase-free, RNase-free, and non-pyrogenic per lot certificate. Vented caps use a 0.2 µm hydrophobic PTFE membrane. As of 2026, NEST Scientific non-treated flasks are stocked by MBP in all four sizes (T25, T75, T175, T225) with vented and plug seal options, in case quantities for PO-based institutional ordering. MBP is a registered vendor for Howard Hughes Medical Institute, Vanderbilt University, and MD Anderson Cancer Center.

Elevate your suspension culture workflows. Contact Molecular Biology Products Inc. for your Non-Treated Cell Culture Flasks today!

FAQ

Non-treated cell culture flasks are sterile polystyrene T-flasks with an unmodified hydrophobic surface (contact angle ~75–90°) that minimizes cell attachment. They are used for suspension mammalian cells (CHO-S, HEK293-F, Jurkat, K562, HL-60), hematopoietic progenitors, hybridomas for monoclonal antibody production, insect cells (Sf9, Hi5), and microbial culture (E. coli, yeast) where surface attachment is undesirable or harmful to cell phenotype.

TC-treated surfaces introduce negatively charged oxygen groups that activate integrin and focal adhesion signaling in hematopoietic cells including Jurkat T-cells and K562 erythroleukemia. This forced adhesion signal induces stress responses, unwanted differentiation, and reduced proliferation in suspension-adapted lines. Non-treated flasks maintain the hydrophobic surface character that keeps hematopoietic cells in suspension and preserves their intended phenotype.

No. Adherent mammalian cell lines such as HeLa, HEK293 (adherent), CHO-K1 (monolayer), Vero, A549, and primary fibroblasts require TC-treated surfaces to attach, spread, and survive. Placing adherent cell lines in non-treated flasks causes cells to remain in suspension without spreading, rapidly lose viability, and die within 24–48 hours. Non-treated flasks are exclusively for cells that grow as suspension cultures or loosely adherent formats.

Yes. Non-treated T-flasks are used for Sf9 (Spodoptera frugiperda) and Hi5 (Trichoplusia ni) insect cell monolayer culture in baculovirus expression vector systems. On non-treated PS, Sf9 cells attach loosely and detach easily with gentle tapping, allowing trypsin-free passaging. TC-treated surfaces cause Sf9 cells to adhere so strongly that recovery without trypsin becomes difficult and recovery efficiency drops significantly.

Non-treated NEST Scientific flasks come with a 0.2 µm hydrophobic PTFE membrane vented cap as the standard CO₂ incubator format. The 0.2 µm pore size blocks bacteria, fungi, and mycoplasma while allowing CO₂ and O₂ gas exchange to maintain medium pH. For static suspension cultures in a CO₂ incubator, always use the vented cap; plug seal caps prevent gas exchange and cause medium acidification within hours.

NEST Scientific catalog 708003 is a non-treated T75 flask with a vented 0.2 µm PTFE membrane cap for CO₂ incubator culture of suspension cells. Catalog 708004 is a non-treated T75 with a plug seal cap, designed for anaerobic culture, transport outside the incubator, and short-term sealed storage of suspension cultures where gas exchange must be prevented. The growth surface, polystyrene material, and sterility specifications are identical between both formats.

Suspension cells in non-treated T-flasks should be passaged when they reach a density of 1–3 × 10⁶ cells/mL (varies by cell line) to avoid nutrient depletion and waste product accumulation. For rapidly growing suspension lines like CHO-S (doubling time ~18–22 hours), passage every 2–3 days maintains optimal viability. Monitor cell viability by trypan blue exclusion or automated cell counter before each passage; viability below 85% indicates culture stress requiring investigation.

MBP stocks NEST Scientific non-treated cell culture flasks in T25, T75, T175, and T225 sizes with vented and plug seal caps, available for PO-based case-quantity ordering with direct human support. MBP is a registered vendor for Howard Hughes Medical Institute, Vanderbilt University, and MD Anderson Cancer Center. Contact MBP for current pricing, case configurations, and lead times.

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