C4-2 cell line人前列腺癌细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

C4-2 cell line人前列腺癌细胞株

Organism Homo sapiens, human
Tissue prostate
Product Format frozen 1.0 mL
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 1
Disease prostate cancer
Gender male
Ethnicity Caucasian
Applications Property of this cell line: Secretion of prostate specific antigen to culture medium. Applications: Prostate cancer tumorigenicity, androgen-independent progression, and bone metastasis
Derivation C4 cells were isolated from a human prostate cancer LNCaP cell subcutaneous xenograft tumor of castrated mouse. LNCaP cells were isolated from a patient lymph node metastasis of prostate cancer.
Comments The human prostatic carcinoma cell line, LNCaP (1 x 106 cells; passage # 29) as described in Horoszewiez, JS et al. Cancer Research 43:1809-1818, 1983; was co-inoculated into an athymic male nude mouse with (1 x 106) human fibroblasts derived from an osteosarcoma (cell line MS). The nude mouse host was castrated after 8 weeks incubation. A tumor specimen was excised after a total of 12 weeks. The C4 cell line constitutes the in vitro cultured subline grown from the murine host’s tumor. When the C4 sub-line was subsequently co-inoculated with MS osteosarcoma fibroblasts in a castrated athymic male nude mouse host for another 12 weeks by the same protocol described above. Prostatic epithelial cells cultured from the resultant tumor in this host constituted the C4-2 subline. Tumorigenicity & Osseous Metastasis: Orthotopic administration of 1 x 106 resuspended C4-2 cells in both intact and castrated athymic male nude mice yielded 100% tumorigenicity (20/20 and 14/14, respectively). Osseous prostate cancer metasteses were detected in both intact and castrated murine hosts (2/20 and 3/14, respectively
Complete Growth Medium The base medium for this cell line is DMEM/F12(4:1). To make the complete medium add: 10% FBS, heat inactivated; 0.100 μg/mL Insulin; 275 ng/mL Triiodothyronine; 88.6 ng/mL apo-Transferrin; 4.9ng/mL d–Biotin; 251.8 ng/mL Adenine. Combine all components, mix well, and sterilize using a 0.22-micron filter. Aseptically dispense 5.7 mL into sterile tubes. Store at -20°C.
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. Corning® T-75 flasks are recommended for subculturing this product.
1.Remove and discard culture medium.
2.Briefly rinse the cell layer with DPBS to remove all traces of serum that contains trypsin inhibitor.
3.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.
4.Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5.Add appropriate aliquots of the cell suspension to new culture vessels.
Cultures can be established between 2.0 x 104 and 3.0 x 104 viable cells/cm2.
6.Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 1.5 X 104 and 3.1 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:8 to 1:10 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation 95% complete growth media + 5% DMSO. Store at liquid nitrogen vapor phase.
Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial Approximately 2 to 3 x 106 cells
Volume 1.0 mL
STR Profile Amelogenin: X,Y
CSF1PO: 9,10,11
D13S317: 10,11
D16S539: 10,11
D5S818: 11,12
D7S820: 9.1,10.3
TH01: 9
TPOX: 8,9
vWA: 16,18
Sterility Tests Bacteria and yeast: No growth
Mycoplasma: No growth
Viability ≥ 50%
Population Doubling Time approximately 24-36 hours
References Thalmann GN, et al. Androgen-independent cancer progression and bone metastasis in the LNCaP model of human prostate cancer. Canc Res 54(10):2577-2581, 1994. PubMed: 8168083

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