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BCRJ Code 0014
Cell Line 3A10
Species Rattus norvegicus
Vulgar Name Rat
Tissue Blood
Cell Type Hybridoma: B Lymphocyte
Morphology Lymphoblast
Growth Properties Suspension
Sex Female
Products Immunoglobulin; monoclonal antibody; TcR murine: ab against; gamma-delta: ab against
Biosafety 1
Addtional Info This hybridoma secretes monoclonal antibody against TcR murine ( gamma-delta molecule
Culture Medium Iscove's Modified Dulbecco's Medium (IMDM) contains 4 mM L-glutamine, 4500 mg/L glucose with fetal bovine serum to a final concentration of 20%.
Subculturing Cultures can be maintained by addition of fresh medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 x 10e5 viable cells/mL. Maintain cultures at a cell concentration between 1 x 10e5 and 1 x 10e6 cells/mL. NOTE: Do not allow the cell concentration to exceed 1 x 10e6 cells/mL.
Subculturing Medium Renewal Every 2 to 3 days
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 Serafini, T., Colamarino, S.A., Leonardo, E.D., Wang, H., Beddington, R., Skarnes, W.C., and Tessier-Lavigne, M. (1996). Netrin-1 is required for commissural axon guidance in the developing vertebrate nervous system. Cell 87, 1001-1014. Begum, S., Komiyama, M., Toyota, N., Obinata, T., Maruyama, K., and Shimada, Y. (1998). Differentiation of muscle-specific proteins in chicken somites as studied by immunofluorescence microscopy. Cell Tissue Res. 293, 305-311. Phelps, P.E., Alijani, A., and Tran, T.S. (1999). Ventrally located commissural neurons express the GABAergic phenotype in developing rat spinal cord. J. Comp. Neurol. 409, 285-298. Perez, S.E., Rebelo, S., and Anderson, D.J. (1999). Early specification of sensory neuron fate revealed by expression and function of neurogenins in the chick embryo. Development 126, 1715-1728. Robb, S.M.C., and Sanchez Alvarado, A. (2002). Identification of immunological reagents for use in the study of freshwater planarians by means of whole-mount immunofluorescence and confocal microscopy. Genesis 32, 293-298. Cao, X., Pfaff, S.L., and Gage, F.H. (2007). A functional study of miR-124 in the developing neural tube. Genes Dev. 21, 531-536. Cao, X., Pfaff, S.L., and Gage, F.H. (2008). YAP regulates neural progenitor cell number via the TEA domain transcription factor. Genes Dev. 22, 3320-3334. Sanchez-Guardado, L.O., Ferran, J.L., Mijares, J., Puelles, L., Rodriguez-Gallardo, L., and Hidalgo-Sanchez, M. (2009). Raldh3 gene expression pattern in the developing chicken inner ear. J. Comp. Neurol. 514, 49-65. Gerhart, J., Scheinfeld, V.L., Milito, T., Pfautz, J., Neely, C., Fisher-Vance, D., Sutter, K., Crawford, M., Knudsen, K., and George Weinstein, M. (2011). Myo/Nog cell regulation of bone morphogenetic protein signaling in the blastocyst is essential for normal morphogenesis and striated muscle lineage specification. Dev. Biol. 359, 12-25.
Depositors Alberto Nobrega, Universidade Federal do Rio de Janeiro