0157 - MA 104

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BCRJ Code 0157
Cell Line MA 104
Species Macaca mulatta
Vulgar Name Monkey; Rhesus Monkey
Tissue Kidney
Morphology Epithelial
Growth Properties Adherent
Derivation Established from an African Green monkey foetal kidney. The cells are highly susceptible to Simian rotavirus SA11
Virus Succeptility: SIMIAN ROTAVIRUS SA11
Biosafety 1
Addtional Info Please note: The species of origin of the MA104 cell line has been shown to be different from that claimed by the originators. Whitaker & Hayward (1985) found the cell line to originate from an African Green monkey and not a Rhesus macaque as was originally claimed. Whitaker AM & Hayward CJ (1985) The characterization of three monkey kidney cell lines. Develop. Biol. Standard. Vol 60. Pp 125 – 131. PMID: 4043530. Abstract: Three monkey kidney cell lines, Vero, GL-V3 and MA-104 were subjected to karyological analysis to determine their chromosomal stability and to confirm their species of origin. Although the lines were shown to be relatively stable throughout all of the passage levels that were tested, the species of origin of one of them was found to be different from that claimed by the originators. This finding was supported by data from isoenzyme studies.
Culture Medium Dulbecco's Modified Eagle's Medium (DMEM) with 1% non-essential amino acids, 2 mM L-glutamine, 1 mM sodium pyruvate, 1.0 g/L glucose and 10% of fetal bovine serum.
Subculturing Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with PBS without calcium and magnesium to remove all traces of serum which contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. 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 Medium Renewal 2 to 3 times per week
Subculturing Subcultivation Ratio 1:3 to 1:10
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 J Gen Virol 1979;43:513; Arch Virol 1981;70:33; Whitaker AM & Hayward CJ (1985) The characterization of three monkey kidney cell lines. Develop. Biol. Standard. Vol 60. Pp 125 – 131. PMID: 4043530.
Depositors Jerson de Lima, Universidade Federal do Rio de Janeiro.
Cellosaurus CVCL_3845