hTERT Human Prostate Cancer-Associated Fibroblasts (CAF)

Catalog Number: HT11
Product Format: Frozen Vial
Cell Number: 1,000,000 cells/vial
Suggested Medium: CAF Growth Medium

General Information

Puromycin-resistant hTERT cells were selected from HPCAF after infection with lentiviruses expressing hTERT.
These Human Prostate CAFs were isolated from prostate tumor tissue. Cells were cultured in T75 flasks pre-coated with AlphaBioCoat and expanded in CAF Growth Medium for 3–7 days. They were harvested at passage 1 and cryopreserved. Each vial contains at least 1,000,000 viable cells.

Product Testing

• Negative for: von Willebrand Factor/Factor VIII, cytokeratin 18, and alpha-smooth muscle actin

• Free from: bacteria, yeast, fungi, and mycoplasma

• CAF Marker Expression: FAP, PDGFR, Vimentin, PDPN, CD70

• Expandable: 3–5 passages at a 1:2 or 1:3 split ratio

Laboratory Applications

• Tumor microenvironment modeling

• Cell interaction and migration studies

• Western blot, PCR, and immunoprecipitation

• Flow cytometry and immunofluorescence

• Development of CAF-based prostate cancer models

Shipping

Frozen vials shipped on dry ice

Post-Thaw Culture Instructions

1. Thawing & Aseptic Handling

   • Thaw the cryovial in a 37°C water bath (~2 minutes) with gentle agitation.

   • Do not submerge the cap.

   • Disinfect the vial using 70% ethanol.

   • Continue under aseptic conditions.

2. Recovery & Seeding

   • Centrifuge cells at 125 × g for 5–10 minutes.

   • Discard supernatant and resuspend in fresh CAF Growth Medium.

   • Coat a T-flask with 6–8 mL AlphaBioCoat for 15 minutes.

   • Rinse with 8 mL 1× PBS and discard.

   • Transfer cell suspension to the coated flask.

3. Culture Conditions

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

   • Pre-incubate growth medium for 15 minutes to allow pH stabilization (target: 7.0–7.6).

Subculturing Instructions

Based on 75 cm² flask volumes. Adjust for different formats.

1. Discard spent medium.

2. Rinse the cell layer with 1× PBS.

3. Add 2.0–3.0 mL Cell Detachment Solution. Incubate at 37°C for 5–15 minutes.

   • Do not shake or tap the flask.

   • Extend incubation if cells are difficult to detach.

4. Pre-coat a new flask with AlphaBioCoat, rinse, and discard PBS.

5. Resuspend cells in 6.0–8.0 mL fresh growth medium.

6. Plate appropriate aliquots into new vessels.

• Subcultivation Ratio: 1:2 to 1:3

• Medium Renewal: Every 3 to 4 days

Cryopreservation Protocol

• Cryopreservation medium: Complete growth medium + 5% (v/v) DMSO

• Catalog #: CGM5

• Freeze at –80°C overnight using an isopropanol-based freezing container

• Transfer to liquid nitrogen for long-term storage

Product Usage

Cells are offered for Research Use Only. Not for Clinical Use.