hTERT Human Lung Fibroblasts Adenocarcinoma

$6,000.00

hTERT-Immortalized Puromycin-Resistant Lung CAFs These genetically engineered lung cancer-associated fibroblasts (CAFs) are derived from primary human lung tumor isolates, immortalized via lentiviral hTERT expression and selected for puromycin resistance to ensure stable, contamination-free cultures. Isolated from lung adenocarcinoma tissue and expanded in AlphaBioCoat-treated T75 flasks using specialized CAF Growth Medium, these cells maintain key tumor microenvironment markers (FAP+/α-SMA+) while offering unlimited proliferation capacity. Cryopreserved at Passage 1, each vial contains ≥1×10⁶ cells/mL with guaranteed viability and sterility (mycoplasma/bacteria-free). Ideal for long-term studies of lung cancer stroma, drug resistance mechanisms, and high-throughput screening applications where genetic manipulation or consistent performance across passages is required.

hTERT-Immortalized Puromycin-Resistant Lung CAFs These genetically engineered lung cancer-associated fibroblasts (CAFs) are derived from primary human lung tumor isolates, immortalized via lentiviral hTERT expression and selected for puromycin resistance to ensure stable, contamination-free cultures. Isolated from lung adenocarcinoma tissue and expanded in AlphaBioCoat-treated T75 flasks using specialized CAF Growth Medium, these cells maintain key tumor microenvironment markers (FAP+/α-SMA+) while offering unlimited proliferation capacity. Cryopreserved at Passage 1, each vial contains ≥1×10⁶ cells/mL with guaranteed viability and sterility (mycoplasma/bacteria-free). Ideal for long-term studies of lung cancer stroma, drug resistance mechanisms, and high-throughput screening applications where genetic manipulation or consistent performance across passages is required.

Product Testing & Quality Control

Human Lung Cancer-Associated Fibroblasts (CAFs) undergo rigorous testing to ensure identity, purity, and performance. These cells are confirmed negative for endothelial and epithelial markers, including von Willebrand Factor (vWF/Factor VIII), Cytokeratin 18, and alpha-smooth muscle actin, helping maintain a pure fibroblast population.

All batches are also tested and verified to be free from bacteria, yeast, fungi, and mycoplasma contamination.

Cells are positive for key CAF markers including:

  • FAP (Fibroblast Activation Protein)

  • PDGFR (Platelet-Derived Growth Factor Receptor)

  • Vimentin

  • PDPN (Podoplanin)

  • CD70

Human Lung CAFs can be expanded for 3–5 passages using a 1:2 to 1:3 split ratio under recommended culture conditions.

Laboratory Applications

Human Lung CAFs are ideal for a wide range of experimental research, including:

  • Cell-cell interaction studies

  • Adhesion assays

  • PCR and Western blot analysis

  • Immunoprecipitation

  • Immunofluorescence and flow cytometry

  • Generation of derivative cell lines for advanced applications

These cells provide an excellent platform for studying the tumor microenvironment and fibrosis in pulmonary oncology research.

Shipping & Handling – Frozen Vials (Dry Ice)

Cells are shipped frozen on dry ice to preserve maximum viability.

Important Guidelines Upon Receipt:

  • Thaw and initiate culture immediately upon arrival.

  • If storage is necessary, keep vials in the liquid nitrogen vapor phase only.
    Do not store at -70°C, as this significantly reduces cell viability.

Thawing Instructions:

  1. Thaw the vial in a 37°C water bath with gentle agitation for ~2 minutes. Avoid submerging the cap or O-ring.

  2. Immediately decontaminate the vial with 70% ethanol and perform all further handling under strict aseptic conditions.

  3. Centrifuge the contents at 125 x g for 5–10 minutes to remove the cryoprotectant. Discard the supernatant and resuspend the cell pellet in fresh, pre-warmed growth medium.

  4. Pre-coat a T-75 flask with 6–8 mL AlphaBioCoat for 15 minutes. Rinse with 8 mL of 1X PBS, discard, and transfer the cells to the flask.

  5. Ensure the growth medium reaches physiological pH (7.0–7.6) by placing the culture vessel in the incubator for 15 minutes prior to cell addition.

  6. Incubate at 37°C in a humidified 5% CO₂ incubator.

Subculturing Protocol

For optimal results, use T-75 flasks and scale appropriately for other vessel sizes.

Note: Avoid agitating the flask while waiting for cells to detach. If necessary, place at 37°C to aid in dissociation.

Steps:

  1. Remove and discard the culture medium.

  2. Rinse briefly with 1X PBS to remove serum that may inhibit dissociation.

  3. Add 2–3 mL of Cell Detachment Solution and monitor under a microscope until cells begin to disperse (5–15 minutes).

  4. Coat a fresh T-flask with AlphaBioCoat (6–8 mL for 15 minutes), rinse with 1X PBS, and discard the rinse.

  5. Resuspend cells in 6–8 mL of complete growth medium and gently pipette to achieve a single-cell suspension.

  6. Plate into new flasks at a 1:2 or 1:3 split ratio.

  7. Replace medium every 3–4 days.

Cryopreservation

For long-term storage, cells should be cryopreserved in complete growth medium supplemented with 5% DMSO. Use standard controlled-rate freezing or a freezing container for best results.