WPE1-NB14

WPE1-NB14 cells belong to a family of cell lines, referred to as the MNU cell lines, which are all derived from RWPE-1 cells after exposure to MNU. The larger family of cell lines, including RWPE-1 cells with a common lineage, mimics multiple steps in progression from normal epithelium to prostatic intra-epithelial neoplasia, and then to invasive cancer. The MNU cell lines, in order of increasing malignancy are: WPE1-NA22 (ATCC CRL-2849), WPE1-NB14 (ATCC CRL-2850), WPE1-NB11 (ATCC CRL-2851), and WPE1-NB26 (ATCC CRL-2852).

WPE1-NB14 cells show moderate invasive ability in the in vitro Boyden chamber invasion assay RefWebber MM, et al. Human cell lines as an in vitro/in vivo model for prostate carcinogenesis and progression. Prostate 47: 1-13, 2001. PubMed: 11304724.

The colony forming efficiency (CFE) of 1.85% and the invasive ability of WPE1-NB14 cells are both greater than those of WPE1-NA22 cells (ATCC CRL-2849) RefWebber MM, et al. Human cell lines as an in vitro/in vivo model for prostate carcinogenesis and progression. Prostate 47: 1-13, 2001. PubMed: 11304724.

The cells form small tumors after subcutaneous injection. The tumors are a little larger than those formed by WPE1-NA22 cells (ATCC CRL-2849).

The depositor report that the parent RWPE-1 cell line (ATCC CRL-11609) was screened for Hepatitis B and C, and human immunodeficiency viruses, and was found to be negative.

1. Check all containers for leakage or breakage.
2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.

1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete growth medium and spin at approximately 125 x g for 5 to 7 minutes. Discard supernatant.
4. Resuspend the cell pellet with the recommended complete growth medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a 25 cm² or a 75 cm² culture flask. It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
5.  Incubate the culture at 37°C in a suitable incubator. A 5% CO₂ in air atmosphere is recommended if using the medium described on this product sheet.

Volumes used in this protocol are for 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note:Subculture cells before they reach confluence. Do not allow cells to become confluent.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS).
3. Add 2.0 to 3.0 mL (to a T-25 flask) or 3.0 to 4.0 ml (to a T-75 flask) of 0.05% Trypsin - 0.53 mM EDTA solution, diluted 1:1 with D-PBS, and place flask in a 37°C incubator for 5 to 8 minutes. Observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 10 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.
4. Add 6.0 to 8.0 mL of 0.1% Soybean Trypsin Inhibitor (or 2% fetal bovine serum in D-PBS), as appropriate, and aspirate cells by gently pipetting.
5. Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 7 minutes.
6. Discard supernatant and resuspend cells in fresh serum-free growth medium. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 2 x 10⁴ to 4 x 10⁴ viable cells/cm² is recommended.
7. Incubate cultures at 37°C. We recommend that you maintain cultures at a cell concentration between 4 x 10³ and 8 x 10⁴ cells/cm².

Note: Cells grown under serum-free or reduced serum conditions may not attach strongly during the 24 hours after subculture and should be disturbed as little as possible during that period.

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

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