Immortalized Human Primary Bronchial Fibroblasts

$3,000.00

Immortalized Human Primary Bronchial Fibroblasts are specialized cells derived from the bronchial submucosa and genetically modified for extended proliferation while maintaining key physiological characteristics. These cells exhibit typical fibroblast morphology and express standard markers including vimentin, collagen I/III, and fibronectin, while testing negative for epithelial (cytokeratin 19) and smooth muscle (α-SMA) contaminants. Certified free of mycoplasma, bacteria, yeast, and fungi, these bronchial fibroblasts demonstrate stable growth for 20+ passages in optimized fibroblast medium when cultured at 37°C/5% CO₂. Particularly valuable for respiratory research, they are ideal for studying airway remodeling, fibrosis mechanisms, and epithelial-mesenchymal interactions in conditions such as asthma, COPD, and pulmonary fibrosis. Supplied cryopreserved at passage 3 with ≥1×10⁶ viable cells/vial, these cells retain responsiveness to TGF-β and PDGF signaling pathways and show consistent doubling times of 24-36 hours during logarithmic growth phase, making them reliable for both short-term experiments and long-term co-culture studies.

Immortalized Human Primary Bronchial Fibroblasts are specialized cells derived from the bronchial submucosa and genetically modified for extended proliferation while maintaining key physiological characteristics. These cells exhibit typical fibroblast morphology and express standard markers including vimentin, collagen I/III, and fibronectin, while testing negative for epithelial (cytokeratin 19) and smooth muscle (α-SMA) contaminants. Certified free of mycoplasma, bacteria, yeast, and fungi, these bronchial fibroblasts demonstrate stable growth for 20+ passages in optimized fibroblast medium when cultured at 37°C/5% CO₂. Particularly valuable for respiratory research, they are ideal for studying airway remodeling, fibrosis mechanisms, and epithelial-mesenchymal interactions in conditions such as asthma, COPD, and pulmonary fibrosis. Supplied cryopreserved at passage 3 with ≥1×10⁶ viable cells/vial, these cells retain responsiveness to TGF-β and PDGF signaling pathways and show consistent doubling times of 24-36 hours during logarithmic growth phase, making them reliable for both short-term experiments and long-term co-culture studies.


Protocol for Culturing Immortalized Human Primary Bronchial Fibroblasts

Materials Required:

  • Cells & Media:

    • Immortalized Human Primary Bronchial Fibroblasts

    • Complete Immortalized Bronchial Fibroblast Growth Medium (IBFGM001)

    • Cryopreservation medium (with DMSO)

  • Solutions & Reagents:

    • Sterile 1X PBS (PBS001, Ca²⁺/Mg²⁺-free)

    • Cell Detachment Solution

    • Neutralization Solution

  • Equipment & Consumables:

    • AlphaBioCoat-coated T25 flasks

    • 37°C, 5% CO₂ humidified incubator

    • Centrifuge (for passaging)

    • Hemocytometer or automated cell counter

    • Liquid nitrogen storage system

1. Thawing Frozen Cells

  1. Pre-warm Medium: Heat IBFGM001 to 37°C.

  2. Thaw Cells:

    • Retrieve cryovial from liquid nitrogen and thaw in a 37°C water bath (1–2 min).

    • Sterilize vial with 70% ethanol.

  3. Dilute & Centrifuge:

    • Transfer cells to a 15 mL conical tube, add 5–10 mL warm medium dropwise.

    • Centrifuge at 200 × g for 5 min.

  4. Plate Cells:

    • Resuspend pellet in 5 mL fresh medium, seed in AlphaBioCoat T25 flask.

    • Incubate at 37°C, 5% CO₂.

2. Routine Maintenance & Passaging

  1. Monitor Growth:

    • Check spindle-shaped morphology daily; passage at 80–90% confluency.

  2. Passaging Steps:

    • Wash cells with 1–2 mL PBS (T25).

    • Add 1–2 mL Detachment Solution, incubate at 37°C for 2–5 min (monitor detachment).

    • Neutralize with 2–3 mL Neutralization Solution, pipet gently.

    • Optional: Centrifuge (200 × g, 5 min) to remove residual enzymes.

  3. Split & Replate:

    • Resuspend in fresh medium, seed at 1:2 to 1:4 ratio.

3. Cryopreservation

  1. Harvest & Count: Follow passaging protocol; adjust density to 1–5 × 10⁶ cells/mL in freezing medium.

  2. Aliquot: Distribute 1 mL/vial into cryovials.

  3. Freeze:

    • Use isopropanol freezing container at –80°C overnight.

    • Transfer to liquid nitrogen for long-term storage.

4. Quality Control & Troubleshooting

✔ Contamination Check: Monitor medium clarity and pH.
✔ Morphology: Healthy cells are elongated/spindle-shaped; rounding indicates stress.
✔ Doubling Time: Expect 24–48 hours; prolonged doubling may signal senescence.
✔ Passage Limit: Use within 20–30 passages for consistent results.

Expected Outcomes

  • Adhesion: Within 24 hours.

  • Proliferation: Doubling time 24–48 hours.

  • Applications:

    • Airway remodeling studies

    • Fibrosis and ECM research

    • Co-culture models (e.g., epithelial-fibroblast interactions)