Human Cell Lines-Cell Lines-InCellGene LLC.-CellLines,Biomedicine reasearch Product,Cell Biology,Cell Culture,293T Cell,Media

Organism	Homo sapiens, human
Tissue	mesothelium
Cell Type	epithelial virus transformed
Product Format	frozen
Morphology	epithelial
Culture Properties	adherent
Biosafety Level	2 [Cells contain polyomavirus DNA sequences] Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease	pleural fluids obtained from non-cancerous individuals.
Applications	The cells can be used to screen chemical and biological agents for activity as growth factor, carcinogens, mutagens, etc.
Storage Conditions	liquid nitrogen vapor phase
Disclosure	This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.

Derivation	The cells were transfected with the pRSV-T plasmid (an SV40 ori- construct containing the SV40 early region and the Rous sarcoma virus long terminal repeat) and cloned. Mesothelial cells were isolated from pleural fluids obtained from non-cancerous individuals.
Tumorigenic	No
Effects	No, The cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Comments	The cells stain positively for vimentin, keratins and SV40 T antigen.

Complete Growth Medium	The base medium for this cell line is Medium 199 containing 1.5 g/L sodium bicarbonate. To make the complete growth medium, add the following components to the base medium: 10% fetal bovine serum (final conc.) 3.3 nM epidermal growth factor (EGF) (final conc.) (do not filter). 400 nM hydrocortisone (final conc.) 870 nM zinc-free bovine insulin (final conc.) 20 mM HEPES (final conc.) The trace elements at the following final concentrations: H2SeO3 0.3869 mg/L (Selenious acid) MnCl2×4H20 0.0198 mg/L (Manganese chloride) Na2SiO3×9H20 14.2100 mg/L (Sodium silicate) (NH4)6Mo7O24×4H20 0.1236 mg/L (Ammonium molybdate) NH4VO3 0.0585 mg/L (Ammonium vanadate) NiSO4×6H20 0.0131 mg/L (Nickle sulfate) SnCl2×2H20 0.0113 mg/L (Tin Chloride) See Methods in Cell Biology, Vol. 21B, pg. 200, 1980 This medium is formulated for use with a 5% CO2 in air atmosphere. ATCC tested fetal bovine serum is available as ATCC Catalog No. 30-2020 (500ml) and ATCC Catalog No. 30-2021 (100ml).
Subculturing	Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. Incubate cultures at 37°C. Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium Renewal: Every 2 to 3 days
Cryopreservation	Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Temperature: 37°C