WPE-stem

WPE-stem cells are loosely adherent and exhibit features characteristic of stem/progenitor cells present in the embryonic urogenital sinus and in adult prostatic epithelium, including p63 and ABCG2.

The cells show high expression of cytokeratin 5 and 14 and MMP-2 but low expression of cytokeratin 18.

They are androgen-independent for growth and survival.

Cells grow in soft agar with a cloning efficiency of 0.9%.

Cells are not tumorigenic in nude mice even at 6 months after injection.

WPE-stem cells were screened for CMV, HBV, HCV, HTLV 1, HTLV 2, HIV 1, HIV 2, JCV, and MoMuLV DNA sequences. Cells were also tested for 25 species of mycoplasma and Acholeplasma laidlawii . Cells tested negative for all of the above. (personal communication from depositor).

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.

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

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 culture medium and spin at approximately 125 x g for 5 to 7 minutes.
4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a pre-coated 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 7.4).
5. Incubate the culture at 37°C in a suitable, humidified incubator. A 5% CO₂ in air atmosphere is recommended if using the medium described on this product.

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

1. Collect spent medium containing viable, floating, single cells, and prostaspheres.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS).
3. 3.0 to 4.0 mL of 0.05% Trypsin - 0.53 mM EDTA solution, diluted 1:1 with D-PBS, and place flask in a 37°C incubator for 3 to 5 minutes. Observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 8 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, add the spent medium containing floating cells, 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 pre-coated culture flasks. An inoculum of 2 X 10³ to 4 X 10³ viable cells/cm² is recommended.
7. Place culture vessels in incubators at 37°C. 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.

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.

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.