C4-2B (ATCC® CRL-3315)

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

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Organism Homo sapiens, human
Tissue prostate
Product Format frozen 1.0 mL
Morphology epithelial-like with thin processes
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 Prostate cancer tumorigenicity, androgen-independent progression, and bone metastasis
Images Micrograph of CRL-3315
Comments Derivative subline of human prostate cancer LNCaP-derived C4-2 cells. 

Origin: The human prostatic carcinoma cell line LNCaP (previously described) was co-inoculated into an athymic male nude mouse with human fibroblasts derived from an osteosarcoma. 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.

To prepare the T-medium supplement:

 Item Name Vendor & Catalog Number   Final Concentration of Media in 1L Amount to prep 1L 
 Insulin  Gibco 12585-014  10 µg/mL  10 mg
 3,3',5-Triiodo-L-thyronine  Sigma T2877  27.3 µg/mL  27.3 mg
 Apo-transferrin (Human)  Sigma T4382  8.8 µg/mL  8.8 mg
 D-Biotin  Sigma 47868  0.488 µg/mL  0.488 mg
 Adenine  Sigma A3159  25 µg/mL  25 mg
 PBS  ATCC 30-2200  N/A  1 L
 BSA (Bovine Serum Albumin)  Sigma  N/A  1 g

Make 100 mL of 0.1% BSA/PBS solution. Combine all components, mix well, and sterilize using a 0.22 micron filter. Aseptically dispense 5.7 mL into sterile tubes.

          
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 supplement below)

To prepare the T-medium supplement:

Prepare 100 mL of a 0.1% BSA (Sigma cat# A8022) in PBS (ATCC 30-2200) solution 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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with DPBS (ATCC 30-2200) to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL 0.01% (w/v) Trypsin- 0.53 mM 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 (ATCC 4-X). Store at liquid nitrogen vapor phase.
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C

To prepare the T-medium supplement:

 Item Name Vendor & Catalog Number   Final Concentration of Media in 1L Amount to prep 1L 
 Insulin  Gibco 12585-014  10 µg/mL  10 mg
 3,3',5-Triiodo-L-thyronine  Sigma T2877  27.3 µg/mL  27.3 mg
 Apo-transferrin (Human)  Sigma T4382  8.8 µg/mL  8.8 mg
 D-Biotin  Sigma 47868  0.488 µg/mL  0.488 mg
 Adenine  Sigma A3159  25 µg/mL  25 mg
 PBS  ATCC 30-2200  N/A  1 L
 BSA (Bovine Serum Albumin)  Sigma  N/A  1 g

Make 100 mL of 0.1% BSA/PBS solution. Combine all components, mix well, and sterilize using a 0.22 micron filter. Aseptically dispense 5.7 mL into sterile tubes.

          
Cells per Vial Approximately 2.0 to 3.0 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: 8.1, 9.1, 10.3
THO1: 9
TPOX: 8, 9
vWA: 16, 18
Sterility Tests Bacteria and yeast: No growth
Mycoplasma: No growth
Viability ≥ 50%
Population Doubling Time 24-36 hours
Name of Depositor K Sherman, University of Texas M.D. Anderson Cancer Center
Year of Origin 1993
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

Thalmann GN, et al. LNCaP progression model of human prostate cancer: androgen-independence and osseous metastasis. Prostate 44(2): 91-103, 2000. PubMed: 10881018

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.
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Other Documentation
Restrictions

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;
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For instructions on how to obtain a license from Institution, please contact the Institution Office of Technology Commercialization via email at researchtools@mdanderson.org.

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

Thalmann GN, et al. LNCaP progression model of human prostate cancer: androgen-independence and osseous metastasis. Prostate 44(2): 91-103, 2000. PubMed: 10881018