P388D1 (IL-1)

Code: 0199
Cell Line: P388D1 (IL-1)
Species: Mus musculus

Vulgar Name: Mouse; Dba/2
Morphology: Macrophage
Disease: Lymphoid Neoplasm
Growth Properties: Adherent
Derivation: P388D1(IL-1) cells were selected from P388D1 cell line for its high production of interleukin-1 (IL-1).
Products: interleukin-1 (interleukin 1, IL-1 or LAF) induced by LPS or phorbol myristic acid (PMA, TPA); lysozyme
Biosafey: 1
Additional info: These cells phagocytose zymosan and latex beads. They are active in antibody dependent cell mediated cytotoxicity systems.
Culture Medium: RPMI 1640 with 2.0 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 10.0 mM HEPES, and 1.0 mM sodium pyruvate, 90%; fetal bovine serum, 10%.
Subculturing: Remove old medium, add fresh, dislodge cells by scraping, and dispense into new flasks. 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.

Medium Renewal: 3 times per week

Subcultivation ratio: 1:4 to 1:8
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: Mizel SB. Physicochemical characterization of lymphocyte-activating factor (LAF). J. Immunol. 122: 2167-2172, 1979. PubMed: 312860 Mizel SB. Biochemical and biological characterization of lymphocyte-activating factor (LAF) produced by the murine macrophage cell line, P388D1. Ann. N.Y. Acad. Sci. 332: 539-549, 1979. PubMed: 231409 Ralph P, Nakoinz I. Antibody-dependent killing of erythrocyte and tumor targets by macrophage-related cell lines: enhancement by PPD and LPS. J. Immunol. 119: 950-954, 1977. PubMed: 894031 Ralph P, Nakoinz I. Direct toxic effects of immunopotentiators on monocytic myelomonocytic, and histiocytic or macrophage tumor cells in culture. Cancer Res. 37: 546-550, 1977. PubMed: 318922 Koren HS, et al. Identification of macrophage-like characteristics in a cultured murine tumor line. J. Immunol. 114: 894-897, 1975. PubMed: 1167564 . Manual of macrophage methodology. New York: Marcel Dekker; 1981.
Depositors: Banco de Células do Rio de Janeiro