hTERT Human Metastatic Ovarian Fibroblasts

$8,000.00

General Information:
Puromycin-resistant hTERT-immortalized cells are derived from Human Metastatic Ovarian Cancer-Associated Fibroblasts (HMOCAF) following infection with lentiviruses encoding the hTERT gene. These fibroblasts are isolated from metastatic ovarian tumor tissue and initially cultured in T-75 tissue culture flasks coated with AlphaBioCoat for 30 minutes. After seeding, the cells are maintained in CAF Growth Medium for 3 to 7 days to promote attachment and growth, followed by expansion. Prior to shipment, cells at passage 1 are harvested, detached, and cryopreserved in vials. Each cryovial contains a minimum of 1,000,000 viable cells per milliliter.

General Information:
Puromycin-resistant hTERT-immortalized cells are derived from Human Metastatic Ovarian Cancer-Associated Fibroblasts (HMOCAF) following infection with lentiviruses encoding the hTERT gene. These fibroblasts are isolated from metastatic ovarian tumor tissue and initially cultured in T-75 tissue culture flasks coated with AlphaBioCoat for 30 minutes. After seeding, the cells are maintained in CAF Growth Medium for 3 to 7 days to promote attachment and growth, followed by expansion. Prior to shipment, cells at passage 1 are harvested, detached, and cryopreserved in vials. Each cryovial contains a minimum of 1,000,000 viable cells per milliliter.

Product Specifications: Human Metastatic Ovarian CAFs

Quality Assurance & Characterization
Our metastatic ovarian CAFs undergo rigorous validation to ensure research reliability:

  • Negative for: Endothelial (von Willebrand Factor/Factor VIII), epithelial (cytokeratin 18), and smooth muscle (α-SMA) markers

  • Positive for: CAF markers FAP, PDGFR, Vimentin, PDPN, and CD70

  • Sterility Testing: Confirmed free of bacteria, yeast, fungi, and mycoplasma

  • Expansion Capacity: Stable growth for 3-5 passages at 1:2–1:3 split ratios

Research Applications
Ideal for studying:

  • Metastatic niche formation and tumor-stroma crosstalk

  • ECM remodeling and therapy resistance mechanisms

  • Techniques: Co-culture assays, flow cytometry, PCR/Western blot, immunoprecipitation

Shipping & Handling

  • Delivered in cryovials on dry ice

  • For optimal viability:

    • Store in liquid nitrogen vapor phase (avoid –70°C)

    • Rapid thaw in 37°C water bath (~2 minutes)

    • Decontaminate vial with 70% ethanol before opening

Cell Recovery Protocol

  1. Centrifuge (125 × g, 5–10 min) to remove cryoprotectant

  2. Resuspend in fresh growth medium

  3. Plate in AlphaBioCoat-treated flasks (6–8 mL coating solution, 15 min)

  4. Maintain at 37°C, 5% CO₂ (pre-equilibrate medium to pH 7.0–7.6)

Subculture Instructions

  1. Detach with 2–3 mL Cell Detachment Solution (5–15 min, 37°C if needed)

  2. Replate at 1:2–1:3 ratio in freshly coated flasks

  3. Refresh medium every 3–4 days

Cryopreservation

  • Use complete growth medium + 5% DMSO

  • Store long-term in liquid nitrogen vapor phase