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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.
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.
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.
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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