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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.
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Karyotype: RWPE-1 cells have a diploid chromosomal ploidy, and show chromosomal variations such as 45, X,-Y, and 51, XY.
- cultureMedium: K-SFM (We do not supply this product; please consider other suppliers. Please let us know if you need further assistance)
- supplements: Supplement the medium with 0.05 mg/mL BPE, 5 ng/mL EGF. The medium should not be entirely filtered. Add BPE and EGF to 10 mL, and after sterile filtering, incorporate this mixture into the medium.
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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