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C4-2 (ATCC® CRL-3314)

Organism: Homo sapiens, human  /  Tissue: prostate  /  Disease: prostate cancer

Permits and Restrictions

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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
Storage Conditions liquid nitrogen vapor phase
Images Cell Micrograph of C4-2, ATCC CRL-3314
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.       

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.
  1. Remove and discard culture medium.
  2. 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.
  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 3x 104 and 6 x 104 viable cells/cm2.
  6. 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
Name of Depositor K Sherman, University of Texas MD Anderson Cancer Center

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

Wu HC, et al. Derivation of androgen-independent human LNCaP prostatic cancer cell sublines: role of bone stromal cells. Int J Canc 57(3):406-412, 1994. PubMed: 9714059

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • For-profit customers must obtain a research use license agreement from the Contributor prior to shipment.
  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation

Unless the Purchaser has a separate license agreement with The University of Texas M. D. Anderson Cancer Center (“Institution”), the ATCC Material is subject to the following restrictions:

  1. The ATCC Material (and any Modifications, Unmodified Derivatives and/or Progeny thereof) may not be used (1) for commercial purposes or Commercial Use by any Purchaser, or (2) by for-profit or commercial entities for any purpose;
  2. Purchaser may not transfer ATCC Materials, Modifications, Unmodified Derivatives, or Progeny to any for-profit entity or commercial entity; and
  3. Purchaser may not use ATCC Materials (or any Modifications, Unmodified Derivatives and/or Progeny thereof) in connection with any research, collaboration or other activities involving a third party that is a commercial entity or for-profit entity.

The restrictions set forth above are in addition to the restrictions set forth in the ATCC Material Transfer Agreement. Capitalized terms have the meanings set forth in the ATCC Material Transfer Agreement unless otherwise indicated above.

For instructions on how to obtain a license from Institution, please contact the Institution Office of Technology Commercialization via email at


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

Wu HC, et al. Derivation of androgen-independent human LNCaP prostatic cancer cell sublines: role of bone stromal cells. Int J Canc 57(3):406-412, 1994. PubMed: 9714059