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

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

Organism  Homo sapiens, human
Tissue  prostate
Product Format  frozen 1.0 mL
Morphology  epithelial-like
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  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 400 mL DMEM (Lonza cat# 12-741) plus 100 mL F12 Medium (Lonza cat# 12-615F). To make the complete medium add the following components:

   56 mL Fetal Bovine Serum (ATCC 30-2020; heat-inactivate before using)
   5.6 mL T-medium supplement (See instructions for preparing T-medium suppplement below)

To prepare the T-medium supplement:

Prepare 100 mL of a 0.1% BSA (Sigma cat# A8022) in PBS (ATCC 30-2200) solution (0.1g of BSA in 100 mL PBS) and add the following components:

          50 mg Insulin (Gibco cat# 12585-014)
          136 ng Triiodo-L-thyronine (Sigma cat# T2877)
          50 mg Transferrin (Sigma cat# T4382)
          2.5 mg D-Biotin (Sigma cat# 47868)
          250 mg Adenine (Sigma cat# A3159)

Combine all components, mix well, filter sterilize the T-medium supplement 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 (catalog #430641) are recommended for subculturing this product.

   Remove and discard culture medium.
   Briefly rinse the cell layer with Dilute 1:5 - 0.05% Trypsin (ATCC catalog # PCS-999-003) in PBS (ATCC catalog # 30-2200)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.
   Add appropriate aliquots of the cell suspension to new culture vessels.
   Cultures can be established between 3x 104 and 6 x 104 viable cells/cm2.
   Incubate cultures at 37°C.

Interval: Maintain cultures at a cell concentration between 3 X 104 and 1 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
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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