Catálogo de Células

0122 - J774 G.8

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BCRJ Code 0122
Cell Line J774 G.8
Species Mus musculus
Vulgar Name Mouse; Balb/C
Tissue Blood
Morphology Macrophage
Disease Sarcoma
Growth Properties Adherent
Sex Female
Products Interleukin-1; IL-1; limphocyte-activating factor; LAF; lisozyme
Biosafety 1
Addtional Info This cell line was adapted to culture from a tumor wich arouse in afemale BALB/c mous. Its growth is inhibited by dextran sulfate, PPD and LP. This cell line exhibits minor cytolysis, but predominanly antibody-dependent phagoccytosis.
Culture Medium Dulbecco's Modified Eagle's Medium (DMEM) with 2 mM L-glutamine, 1.0 g/L glucose and 10% of fetal bovine serum.
Subculturing remove old medium, add fresh, disloge cells by scraping, and dispensse 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.
Subculturing Subcultivation Ratio io 1:3 to 1:6 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: It is strongly recommended to always wear protective gloves, clothing, and a full-face mask when handling frozen vials. Some vials may leak when submerged in liquid nitrogen, allowing nitrogen to slowly enter the vial. Upon thawing, the conversion of liquid nitrogen back to its gas phase may cause the vial to explode or eject its cap with significant force, creating flying debris.
  1. Thaw the vial by gently agitating it in a 37°C water bath. To minimize contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as its contents are thawed and decontaminate it by dipping in or spraying with 70% ethanol. From this point, all operations must be performed under strict aseptic conditions.
  3. For cells sensitive to DMSO, it is recommended to remove the cryoprotective agent immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL of complete culture medium and centrifuge at approximately 125 × g for 5 to 7 minutes.
  4. Discard the supernatant and resuspend the cell pellet in the recommended complete medium (see specific batch information for the appropriate dilution ratio).
  5. Incubate the culture under appropriate atmospheric and temperature conditions (see "Culture Conditions" for this cell line).

NOTE: It is important to avoid excessive alkalinity of the medium during cell recovery. To minimize this risk, it is recommended to place the culture vessel containing the growth medium in the incubator for at least 15 minutes before adding the vial contents. This allows the medium to stabilize at its normal pH (7.0 to 7.6).
References Nature 257:393-394, 1975; Cell. Immunol. 90: 339-357, 1985.
Depositors Wanderley de Souza-Universidade Federal do Rio de Janeiro.
Cellosaurus CVCL_HA26
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