1. Real Time PCR was used to quantify the transgene expression
Cell Type
Immortalized Cells
Biosafety Level
2
Disclaimer
1. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 5. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
PriCoat?T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture.?rigrow X Series Medium (TM0600), 37.0?, 5% CO?.?ote:?riGrow X Series Medium for T0600 (Cat. No. TM0600) is serum-free. Thaw cells in media supplemented with 10% FBS to promote attachment to the vessel, then change thawing media to serum-free TM0600 media within 24 hours, once attachment is observed.Cells are sensitive to trypsin; 1/10 diluted 0.25% Trypsin (TM050) or?entle Dissociation Solution (TM080)?s recommended for subculture procedures.
Quantity
1x106 cells / 1.0 ml
Tissue
Prostate
Morphology
Epithelial
Notes
1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130?, preferably in liquid nitrogen vapor phase storage, until ready for use. To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism
Human (H. sapiens)
Species
Human (H. sapiens)
Propagation
Use of PriCoatTM ?T25 Flasks (G299) or Applied CellExtracellular Matrix (G422) is required for cell adhesion to theculture vessels. Grow cells in ECM-coated culture vessels unless otherwise specified in the PropagationRequirements below.The base medium for this cell line is MCDB 153 medium (Sigma). To makethe complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)*to a final concentration of 1%, Penicillin/Streptomycin Solution (G255) to a finalconcentration of 1%, MEM Non-essential Amino Acid Solution to a final concentration of 1X, bovine pituitaryextract (Gibco) to a final concentration of 50 mug/mL, insulin (Sigma) to a final concentration of 5 mug/mL,transferrin (Sigma) to a final concentration of 5 mug/mL, EGF (Sigma) to a final concentration of 5 ng/mL,hydrocortisone to a final concentration of 200 nM, triiodothyronine (Sigma) to a final concentration of 10 nM,testosterone (Sigma) to a final concentration of 10 nM and sodium selenite (Sigma) to a final concentration of 5ng/mL.Change media every 2-3 days.Carbon dioxide(CO2): 5%, Temperature: 37.0°C.* Do not useheat-inactivated FBS for cell culture unless specified otherwise.
Cryopreservation
Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Quality Control
1) Western blot; 2) qRT-PCR; 3) Three-dimensional cell culture
Applications Range
Research Use Only.
Shipping
Ship with dry ice.
Product Format
Frozen
Product Use
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.
Storage
Vapor phase of liquid nitrogen, or below -130?.
Thawing Protocol
1. Thaw cells quickly in a 37? water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media supplemented with 10% FBS. Centrifuge cells at 125xg for 5-7 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. 5. Incubate the cells at the recommended conditions.
Subculture Protocol
Cells are sensitive to trypsin; 1/10 diluted 0.25% Trypsin (TM050) or Gentle Dissociation Solution (TM080)?s recommended for subculture procedures.?Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 1/10 diluted 0.25% Trypsin-EDTA or?entle Dissociation Solution (TM080) to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37?, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA or Gentle Dissociation Solution (TM080) by adding an equal volume of the complete growth media supplemented with 10% FBS into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions.
Reviews of Immortalized Human Prostate Epithelial Cells