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C4-2B is a cell line with epithelial-like morphology that was derived from human prostate cancer LNCaP-derived C4-2 cells in 1993. LNCaP cells were isolated from the prostate of a White male with prostate cancer. It was deposited by the University of Texas MD Anderson Cancer Center and has applications for cancer research.
Product category
Human cells
Homo sapiens, human
epithelial-like with thin processes
Prostate Cancer
3D cell culture
Cancer research
Product format
Storage conditions
Vapor phase of liquid nitrogen
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ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information


Specific applications
Prostate cancer tumorigenicity, androgen-independent progression, and bone metastasis


Cells per vial
Approximately 2.0 to 5.0 x 106
1.0 mL
Growth properties
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.


Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
The base medium for this cell line is DMEM/F12(4:1). To make the complete medium add: 10% FBS (ATCC 30-2020), 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.
95% Air, 5% CO2
Handling procedure

To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.  

  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  3. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.
  4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).
  5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.


Subculturing procedure
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 of Trypsin-EDTA (ATCC 30-2101) 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
Reagents for cryopreservation
95% complete growth media + 5% DMSO (ATCC 4-X). Store at liquid nitrogen vapor phase.

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Population doubling time
Approximately 24 to 36 hrs
STR profiling
Amelogenin: X,Y
CSF1PO: 9,10,11
D13S317: 10,11
D16S539: 10,11
D18S51: 10,11,12
D21S11: 28,32.2
D3S1358: 14,15,16
D5S818: 11,12
D7S820: 8.1,9.1,10.3
D8S1179: 12,14
FGA: 19,20
Penta_E: 12,16,17
TH01: 9
TPOX: 8,9
vWA: 16,18
Penta_D: 12,13
D19S433: 13,13.2,15
D2S1338: 16,17
≥ 50%


K Sherman, University of Texas M.D. Anderson Cancer Center
Year of origin

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.


This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at

Permits & Restrictions

For-profit Research Use License from MD Anderson Cancer Center

For-profit Organizations: For every order of this item, you must work directly with the contributor, MD Anderson Cancer Center to (i) negotiate a research-use license, and/or (ii) have the contributor provide authorization to ATCC to ship this item under your existing license. We cannot ship this item until we receive communication directly from MD Anderson Cancer Center that we are authorized to ship each order.

We are providing the following contact information, but this information may change without notice:
M.D. Anderson Cancer Center
Office of Technology Commercialization
Email: [email protected]

Once ATCC has received authorization from MD Anderson Cancer Center, your order will be reviewed, and this item will be released for shipment if all requirements are met. If you need assistance with your order, please contact our Customer Care team or your applicable distributor.

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.


Frequently Asked Questions


Curated Citations

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

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