RWPE-1 Cell Line

Brief Introduction
Product List

General Information
Organism	Homo sapiens, human
Cell Line Description	RWPE-1 is an immortalized human prostate epithelial cell line established by Maria M. Webber and colleagues in 1997. The cell line was derived from the peripheral zone of the prostate of a 54-year-old Caucasian male donor; histological examination confirmed the tissue to be normal and non-malignant. Researchers achieved immortalization of the primary epithelial cells by transfecting them with a plasmid carrying a single copy of the human papillomavirus type 18 (HPV-18) genome. RWPE-1 cells grow as an adherent monolayer and exhibit the characteristic cobblestone morphology of epithelial cells. In the fields of urology, oncology, and toxicology, RWPE-1 is globally recognized as the gold-standard model for non-tumorigenic human prostate epithelial cells. Serving as a vital benchmark model for normal prostate biology, it is widely used to study cell differentiation, prostate carcinogenesis, chemical toxicology, and host-pathogen interactions.
Tissue	Prostate
Cell Type	Epithelial cell
Ethnicity	Caucasian
Disease	Normal / Non-malignant tissue origin (Immortalized engineered line)
Morphology	Epithelial-like
Gender	Male
Age	54 years
Product Format	Frozen
Growth Mode	Adherent
Biosafety Level	2 (BSL-2 is required globally because the line was immortalized using a retroviral vector harboring HPV-18 viral DNA sequences, which are stably integrated into the host genome)
Applications	1. An in vitro human "gold standard" control model representing normal prostate epithelial cells for prostate cancer research; 2. Directly modeling the multi-stage process of prostate cancer development through chemical transformation or oncogene transfection (e.g., establishing the tumorigenic, Ki-ras-transformed RWPE-2 cell line); 3. Elucidating the physiological mechanisms of 3D acinar morphogenesis, cell-matrix interactions, and glandular polarity formation using Matrigel matrices; 4. Investigating androgen receptor (AR) signaling, secretion kinetics of prostate-specific antigen (PSA), and hormone-mediated transcriptional activation; 5. Conducting toxicological and pharmacological analyses to evaluate the safety, uptake, and cellular effects of environmental factors and chemopreventive agents.
Shipped In	Dry ice
Storage Temperature	−196°C
Characteristics
Tumorigenic	No, non-tumorigenic.
Transformant	Human Papillomavirus type 18 (HPV-18). Stably contains a single, replication-deficient integrated copy of the HPV-18 genome.
Acinar Morphogenesis	They retain exceptional and highly specific functional characteristics. When cultured in a three-dimensional (3D) Matrigel matrix or co-cultured with stromal cell lines (such as WPMY-1), RWPE-1 cells spontaneously organize into polarized, hollow, spherical acinar structures; upon androgen stimulation, they actively secrete prostate-specific antigen (PSA) into the acinar lumen.
Phenotypic Markers	Co-expresses both luminal-associated cytokeratins (CK8 and CK18) and basal-associated cytokeratins (CK5 and CK14), reflecting an intermediate or progenitor-like prostatic epithelial phenotype. Expresses functional Androgen Receptors (AR), though at lower levels compared to advanced, cancer-derived lines like LNCaP.
Hormone & Growth Factor Response	They exhibit a strong, dose-dependent proliferative response when exposed to epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF); their proliferation is physiologically inhibited by transforming growth factor-beta (TGF-β).
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Completely remove and discard the used complete culture medium from the culture vessel. 2. Gently rinse the cell monolayer with sterile PBS or Ca2+/Mg2+-free Dulbecco’s PBS (D-PBS) to remove residual serum or divalent cations. 3. Add 2.0 to 3.0 mL of 0.05% trypsin-0.53 mM EDTA solution (typically diluted 1:1 with D-PBS) to the T-25 flask, ensuring the cell layer is completely covered. 4. Incubate the flask at 37°C for 5 to 10 minutes; monitor closely under an inverted microscope until the cell layer has completely detached. Key measure to prevent clumping: Do not tap or shake the flask while waiting for the cells to detach, as this can cause severe cell aggregation. 5. Immediately add 6.0 to 8.0 mL of 0.1% soybean trypsin inhibitor or complete culture medium (D-PBS supplemented with 2% FBS) to neutralize trypsin activity. Note: Since RWPE-1 cells are cultured under serum-free conditions, avoid standard serum-based trypsin neutralization methods to maintain the serum-free environment; the use of a trypsin inhibitor is strongly recommended. 6. Gently pipette the cell suspension up and down to disperse the cells, centrifuge at approximately 125 × g for 5 to 7 minutes, carefully aspirate the supernatant, gently resuspend the cell pellet in fresh complete culture medium, and distribute the suspension into new culture vessels.
Thawing Protocol Note	RWPE-1 cells are highly sensitive to drastic pH fluctuations during the early stages of recovery from cryopreservation. Before adding the thawed cell suspension, the culture vessel containing fresh complete growth medium should be placed in a 5% CO2 incubator for at least 15 minutes to allow the medium to equilibrate to a physiological pH range (7.0 to 7.6).
Medium Renewal	Every 2 days (48 hours)
Subcultivation Ratio	Split confluent cultures (80% density) at a standard ratio of 1:3 to 1:5.
Culture Conditions	Atmosphere: Air, 95%; CO2, 5%; Temperature: 37°C

Cat.No.	Product Name	Price
CSC-RR00827	Luciferase Reporter Cell Line - RWPE-1	Inquiry
CSC-RR00828	GFP Reporter Cell Line - RWPE-1	Inquiry

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* For research use only. Not intended for any clinical use.

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