0267 - RD-ES

BCRJ Code 0267
Cell Line RD-ES
Species Homo sapiens
Vulgar Name Human
Tissue Bone
Morphology Epithelial
Disease Ewing'S Sarcoma
Growth Properties Mixed: Adherent And Clusters In Suspension
Sex Male
Age/Ethinicity 19 Year / Caucasian
Derivation The cell line was initiated by G. Marshall and M. Kirchen from a primary osseous Ewings sarcoma of the humerus.
DNA Profile Amelogenin: X,Y CSF1PO:11 D13S317: 12,11 D16S539:11,9 D5S818:11 D7S820:10 THO1:7 TPOX:11,9 vWA:17 D2S1338:20,19 D19S433: 14,13 FGA:25,21 D3S1358:15 D18S51: 18,14 D8S1179: 13 D21S11: 28
Products Antigen Expression: Blood type B; Rh+
Biosafety 1
Addtional Info Ultrastructurally the cells exhibit primitive cell junctions, possess glycogen pools and are 20 to 25 microns in diameter. The cells grow as a loosely attached monolayer in small clusters of 5 to 10 cells.
Culture Medium RPMI-1640 medium modified to contain 2 mM L-glutamine, 4500 mg/L glucose and 10% of fetal bovine serum.
Subculturing Shake the flask after removing most of the medium. Add fresh medium and transfer to new flasks.
Subculturing Medium Renewal 2 to 3 times a week
Subculturing Subcultivation Ratio 1:3 to 1:8 is recommended
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 Hay, R. J., Caputo, J.L., and Macy, M. L., Eds (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC
Depositors Caroline Brunetto - Instituto do Câncer Infantil do RS