RWPE-1

The RWPE-1 cell line, derived from the prostate epithelium of a 54-year-old Caucasian male with no evidence of prostate cancer, is a valuable resource in biomedical research, particularly for studies on prostate biology and cancer. These epithelial cells, characterized by their adherence growth properties and typical epithelial morphology, were immortalized using a replication-deficient retrovirus that carries the E7 gene from human papillomavirus 18 (HPV-18), which inactivates the retinoblastoma protein and promotes cellular immortalization.

RWPE-1 cells, originating from a normal human prostate, are utilized in prostate cancer research, though their androgen receptor expression is relatively modest, especially when compared to tumorigenic prostate cancer-derived cell lines. The epithelial cell line RWPE-1 expresses cytokeratins 8 and 18, which confirm their epithelial lineage. While RWPE-1 cells do express tumor suppressors such as p53 and pRB, reflecting their non-tumorigenic nature, the expression of prostate-specific markers like Kallikrein 3 (KLK3) or PSA is generally low or absent under standard culture conditions.

In 3D cultures, such as those formed in Matrigel, human cells RWPE-1 can organize into acinar structures reminiscent of normal prostate architecture. When it comes to the secretion of PSA (Prostate-Specific Antigen) in response to androgen stimulation, RWPE-1 cells show a less pronounced reaction compared to prostate cancer cell lines. Therefore, while RWPE-1 cells offer a valuable model for understanding the baseline properties of normal prostate epithelial cells.

RWPE-1's non-tumorigenic nature serves as a model for studying the transition to tumorigenic transformation and the dynamics of cancer cells, including metastatic prostate cancer cells and prostate carcinogenesis. The inclusion of factors like EGF and growth hormone in culture conditions can further elucidate the pathways involved in prostatic hyperplasia and the progression towards prostate cancer. In summary, RWPE-1 cells facilitate a comprehensive understanding of prostate cancer, from its initiation in prostatic cell lines to its manifestation in prostate cancer patients.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.