P3HR-1



BCRJ code:
0338
Cell Line:
P3HR-1
Species:
Homo sapiens

Vulgar Name: Human
Tissue:
Lymph
Morphology:
Lymphoblast
Growth Properties:
Suspension
Biosafety:
2
Culture Medium:
RPMI 1640 medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10 mM HEPES, and 1.0 mM sodium pyruvate supplemented with: 10% fetal bovine serum.
Subculturing:
Cultures can be maintained by the addition of fresh medium or replacement of medium.
Alternatively, cultures can be established by centrifugation with subsequent resuspension.
Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels; Maintain at at 3-9 x 10e5 cells/cm2. Place culture vessels in incubator at 37°C.

NOTE: Do not allow the cell concentration to exceed 1 x 10e6 cells/mL.

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.

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:
J Virol 1967;1:1045; J Nat Cancer Inst 1980;64:725
Depositors:
Ana Luiza Monteiro Wanzeller - Instituto Evandro Chagas