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

Organism	Homo sapiens, human
Tissue	prostate; derived from metastatic site: bone
Product Format	frozen
Morphology	epithelial
Culture Properties	adherent
Biosafety Level	1 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	adenocarcinoma
Age	63 years old adult
Gender	male
Ethnicity	Black
Applications	Cells from this cell line produce tumors in nude mice when injected either subcutaneously or orthotopically (intraprostatic). This cell line is suitable for studying cell growth regulation by androgens.
Storage Conditions	liquid nitrogen vapor phase

Derivation	MDA PCa 2b was established from a bone metastasis of 63 year old Black male with androgen-independent adenocarcinoma of the prostate.
Clinical Data	male 63 years old adult Black
Antigen Expression	prostate specific antigen (PSA)
Receptor Expression	androgen receptor, expressed
Tumorigenic	Yes
Effects	Yes, forms tumors in nude mice
Comments	The cell line expresses prostate specific antigen (PSA) and androgen receptor, grows in vitro and in vivo, and is androgen sensitive.

Complete Growth Medium	The base medium for this cell line is F-12K Medium (ATCC 30-2004). To make the complete growth medium, add the following components to the base medium: non heat-inactivated fetal bovine serum (FBS; ATCC 30-2020) to a final concentration of 20% 25 ng/ml cholera toxin (Sigma cat# C8052) 10 ng/ml mouse Epidermal Growth Factor (Corning cat# 354010) 0.005 mM phosphoethanolamine (Sigma cat# P0503) 100 pg/ml hydrocortisone (Sigma cat# H0135) 45 nM sodium selenite (Sigma cat# 9133) 0.005 mg/ml human recombinant insulin (Life Technologies cat# 12585-014) Do not filter complete medium.
Subculturing	Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium 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 Subculture Ratio: 1:2 to 1:4 Medium Renewal: Every 2 to 3 days. 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.
Cryopreservation	Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Temperature: 37°C Atmosphere: air, 95%; carbon dioxide (CO2), 5%