0397 - TPC-1

BCRJ Code 0397
Cell Line TPC-1
Species Homo sapiens
Vulgar Name Human
Tissue Thyroid gland papillary
Cell Type Epithelial
Morphology Epithelial
Disease Carcinoma
Growth Properties Adherent
Sex Female
Age/Ethinicity Adult /
Products Pax8
Biosafety 1
Culture Medium RPMI-1640 medium modified to contain 2 mM L-glutamine, 4500 mg/L glucose and fetal bovine serum to a final concentration of 10%.
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. Remove and discard culture medium. Briefly rinse the cell layer with PBS without calcium and magnesium 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 the 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. Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. Place culture vessels in incubators at 37°C. NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010.
Subculturing Medium Renewal twice/week
Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Cryopreservation 95% FBS + 5% DMSO (Dimethyl sulfoxide)
Thawing Frozen Cells SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials 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 vessel exploding or blowing off its cap with dangerous force creating flying debris. 1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring 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. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. 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.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line). NOTE: 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 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).
References PubMed=2823470; DOI=10.1016/0042-6822(87)90171-1 - Tanaka J., Ogura T., Sato H., Hatano M. Establishment and biological characteriz ation of an in vitro human cytomegalovirus latency model. Virology 161:62 -72(1987) PubMed=2516841; DOI=10.1111/j.1349-7006.1989.tb01645.x Ishizaka Y., Itoh F., Tahira T., Ikeda I., Ogura T., Sugimura T., N agao M. Presence of aberrant transcripts of ret proto-oncogene in a human papi llary thyroid carcinoma cell line. Jpn. J. Cancer Res . 80:1149-1152(1989) PubMed=11439348; DOI=10.1038/sj.onc.1204531 Frasca F., Vigneri P., Vella V., Vigneri R., Wang J.Y .J. Tyrosine kinase inhibitor STI571 enhances thyroid ca ncer cell motile response to hepatocyte growth factor. Oncogene 20:3845-3856( 2001) PubMed=17725429; DOI=10.1089/thy.2007.0097 Meireles A.M., Preto A., Rocha A.S., Rebocho A.P ., Maximo V., Pereira-Castro I., Moreira S., Feijao T., Botelho T., Marques R., Trovisco V., Cirnes L., Alves C., Velho S., Soares P., Sobrinho-Simoes M. Molecular and genotypic characteri zation of human thyroid follicular cell carcinomaderived cell lines. Thyroid 17:707-71 5(2007) PubMed=17804723; DOI=10.1158/0008-5472.CAN-06-4026 van Staveren W.C.G., Solis D.W., Delys L., Duprez L., Andry G., Franc B., Thomas G., Libert F., Dumont J.E., Detours V., Maenhaut C. Human thyroid tumor cell lines derived from differen t tumor types present a common dedifferentiated phenotype. Cancer Res. 67:8113-8120( 2007) PubMed=18713817; DOI=10.1210/jc.2008-1102 Schweppe R.E., Klopper J.P., Korch C., Pugazhe nthi U., Benezra M., Knauf J.A., Fagin J.A., Marlow L.A., Copland J.A., Smallridge R.C., Haugen B.R. Deoxyribonucleic acid profiling analysis of 40 human thyroid cancer c ell lines reveals cross-contamination resulting in cell line redundancy and misidentification. J. Clin. Endocrinol. Metab. 93:4331-4341(2008) PubMed=19087340; DOI=10.1186/1471-2407-8-371 Ribeiro F.R., Meireles A.M., Rocha A.S., Teixeira M.R . Conventional and molecular cytogenetics of human no n-medullary thyroid carcinoma: characterization of eight cell line models and review of the literature on clinical samples. BMC Can cer 8:371-371(2008) PubMed=21868764; DOI=10.1158/1078-0432.CCR-11-0690 Zhao M., Sano D., Pickering C.R., Jasser S.A., Henderson Y .C., Clayman G.L., Sturgis E.M., Ow T.J., Lotan R., Carey T.E., Sacks P.G., Grandis J.R., Sidransky D., Heldin N.-E., Myers J.N. Assembly and initial cha racterization of a panel of 85 genomically validated cell lines from diverse head and neck tumor sites. Clin. Cancer Res. 17:7248-7264(2011) PubMed=22087789; DOI=10.1186/1755-8166-4-26 Maric I., Viaggi S., Caria P., Frau D.V., Degan P., Va nni R. Centrosomal and mitotic abnormalities in cell lines derived from papillary thyroid cancer harboring specific gene alterations. Mol. Cytogenet. 4:26-26(2011) PubMed=23162534; DOI=10.3389/fendo.2012.00133 Saiselet M., Floor S., Tarabichi M., Dom G., Hebrant A ., van Staveren W.C.G., Maenhaut C. Thyroid cance r cell lines: an overview. Front. Endocrinol. 3:133-133(2012) PubMed=23833040; DOI=10.1210/jc.2013-2383 Landa I., Ganly I., Chan T.A., Mitsutake N., Mats use M., Ibrahimpasic T., Ghossein R.A., Fagin J.A. Frequent somat ic TERT promoter mutations in thyroid cancer: higher prevalence in advanced forms of the disease. J. Clin. Endocrinol. Metab. 98:E 1562-E1566(2013)
Depositors Etel Rodrigues Pereira Gimba - Instituto Nacional de Câncer