RWPE-1 Cells
Essential facts about RWPE-1 cells
Description | 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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Organism | Human |
Tissue | Prostate |
Synonyms | RWPE1 |
Details
Age | 54 years |
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Gender | Male |
Ethnicity | Caucasian |
Morphology | Epithelial |
Cell type | Epithelial cell of prostate |
Growth properties | Adherent |
Documentation about RWPE-1 epithelial cells
Citation | RWPE-1 (Cytion catalog number 305217) |
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Biosafety level | 2 |
Genotype
Karyotype | RWPE-1 cells have a diploid chromosomal ploidy, and show chromosomal variations such as 45, X,-Y, and 51, XY. |
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Handling
Culture Medium | K-SFM (We do not supply this product; please consider other suppliers. Please let us know if you need further assistance) |
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Medium 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. |
Passaging solution | 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. |
Freeze medium | CM-1 (Cytion catalog number 800100) |
Handling of cryopreserved cultures |
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Quality verification of RWPE-1 prostate cell lines
Sterility | 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. |
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STR profile |
Amelogenin: x,y
CSF1PO: 13
D13S317: 8,14
D16S539: 9,11
D5S818: 12,15
D7S820: 10,11
TH01: 8,9.3
TPOX: 8,11
vWA: 14,18
D3S1358: 15,16
D21S11: 29,31
D18S51: 14,16
Penta E: 5,12
Penta D: 10,13
D8S1179: 10,14
FGA: 24,25
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Required products
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.