Immortalized Human Primary Prostate Fibroblasts

$3,000.00

Immortalized Human Primary Prostate Fibroblasts are stromal cells derived from prostate tissue and genetically modified to maintain long-term proliferation while retaining key functional properties. These cells exhibit characteristic spindle-shaped morphology and express standard fibroblast markers including vimentin, fibroblast activation protein (FAP), and α-smooth muscle actin (α-SMA), while testing negative for epithelial (cytokeratin 8/18) and endothelial (CD31) contaminants. Certified free of mycoplasma, bacteria, and fungal contamination, these prostate fibroblasts demonstrate stable growth for 30+ passages in specialized fibroblast medium when cultured at 37°C/5% CO₂. Particularly valuable for urological research, they are ideal for studying prostate cancer stroma interactions, benign prostatic hyperplasia (BPH), and fibrotic remodeling processes. Supplied cryopreserved at passage 4 with ≥1×10⁶ viable cells/vial, these fibroblasts maintain functional responsiveness to androgen signaling and show consistent doubling times of 24-36 hours. Their preserved capacity for extracellular matrix production and cytokine secretion makes them excellent for modeling prostate tumor microenvironments, stromal-epithelial crosstalk studies, and therapeutic screening applications targeting prostate stromal biology. The cells retain their ability to modulate prostate cancer cell behavior, making them particularly suitable for investigating tumor-stroma interactions in prostate cancer progression and metastasis.

Immortalized Human Primary Prostate Fibroblasts are stromal cells derived from prostate tissue and genetically modified to maintain long-term proliferation while retaining key functional properties. These cells exhibit characteristic spindle-shaped morphology and express standard fibroblast markers including vimentin, fibroblast activation protein (FAP), and α-smooth muscle actin (α-SMA), while testing negative for epithelial (cytokeratin 8/18) and endothelial (CD31) contaminants. Certified free of mycoplasma, bacteria, and fungal contamination, these prostate fibroblasts demonstrate stable growth for 30+ passages in specialized fibroblast medium when cultured at 37°C/5% CO₂. Particularly valuable for urological research, they are ideal for studying prostate cancer stroma interactions, benign prostatic hyperplasia (BPH), and fibrotic remodeling processes. Supplied cryopreserved at passage 4 with ≥1×10⁶ viable cells/vial, these fibroblasts maintain functional responsiveness to androgen signaling and show consistent doubling times of 24-36 hours. Their preserved capacity for extracellular matrix production and cytokine secretion makes them excellent for modeling prostate tumor microenvironments, stromal-epithelial crosstalk studies, and therapeutic screening applications targeting prostate stromal biology. The cells retain their ability to modulate prostate cancer cell behavior, making them particularly suitable for investigating tumor-stroma interactions in prostate cancer progression and metastasis.

Materials Required

  • Immortalized Human Primary Prostate Fibroblasts

  • Complete Immortalized Prostate Fibroblast Growth Medium (IPFGM002)

  • Sterile Phosphate-Buffered Saline (1× PBS; PBS001)

  • Cell Detachment Solution

  • Alphabiocoat-Coated Tissue Culture Flasks (T25)

  • 37°C Humidified Incubator with 5% CO₂

  • Centrifuge (optional, for passaging)

  • Hemocytometer or Automated Cell Counter

  • Cryopreservation Medium

  • Liquid Nitrogen Storage System (for long-term storage)

Protocol

1. Thawing Frozen Cells

  • Prepare Medium: Warm the complete prostate fibroblast growth medium to 37°C.

  • Thaw Cryovial: Quickly thaw the vial in a 37°C water bath for 1–2 minutes. Once thawed, disinfect the exterior with 70% ethanol.

  • Transfer to Tube: Transfer the cell suspension to a 15 mL conical tube. Slowly add 5–10 mL of pre-warmed medium dropwise to minimize osmotic shock.

  • Centrifuge: Spin at 200 × g for 5 minutes to pellet the cells.

  • Resuspend & Seed: Discard the supernatant, resuspend the cell pellet in fresh growth medium, and transfer to a pre-coated T25 Alphabiocoat flask.

  • Incubate: Place the flask in a 37°C, 5% CO₂ humidified incubator.

2. Routine Maintenance & Passaging

  • Daily Monitoring: Examine cultures under a microscope. Healthy prostate fibroblasts should exhibit a spindle-shaped morphology and reach 80–90% confluency before passaging.

  • Medium Change: Refresh the culture medium every 2–3 days or as needed.

  • Passaging Procedure:

    • Aspirate the old medium and gently rinse cells with 1–2 mL of sterile PBS.

    • Add 1–2 mL of Cell Detachment Solution and incubate for 2–5 minutes at 37°C. Observe detachment under the microscope.

    • Neutralize by adding 2–3 mL of Neutralizing Solution.

    • Gently pipette to achieve a single-cell suspension.

    • Optional: Centrifuge at 200 × g for 5 minutes to remove any residual enzyme.

    • Resuspend in fresh medium and reseed at a 1:2 to 1:4 split ratio, depending on growth rate.

3. Cryopreservation

  • Harvest Cells: Detach and collect cells following steps 2.2–2.5.

  • Count Cells: Use a hemocytometer or automated counter to assess cell density.

  • Prepare Suspension: Resuspend cells in cryopreservation medium at 1–5 × 10⁶ cells/mL.

  • Aliquot: Dispense 1 mL of the suspension into sterile cryovials.

  • Freeze Gradually: Place cryovials in an isopropanol-based freezing container and store at –80°C overnight.

  • Transfer to Liquid Nitrogen: Move vials to liquid nitrogen for long-term storage the following day.

4. Quality Control & Troubleshooting

  • Contamination Monitoring: Routinely inspect cultures for signs of contamination, such as turbidity or color changes in the medium.

  • Morphology Assessment: Maintain spindle-shaped morphology. Rounding or clumping may indicate cellular stress or overgrowth.

  • Doubling Time: Healthy immortalized prostate fibroblasts should double every 24–48 hours under optimal conditions.

Expected Results

  • Adhesion: Cells should attach within 24 hours post-thaw or seeding.

  • Proliferation: Doubling typically occurs every 24–48 hours.

  • Lifespan: Cells can generally be passaged 20–30 times before signs of senescence, assuming they are not fully immortalized.