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LLC-MK2; Kidney; Rhesus Monkey (Macaca mulatta) Item ID: ATCCCL7

Permits and Restrictions

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Organism Macaca mulatta, monkey, rhesus
Tissue kidney
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease Normal
Age adult
Storage Conditions liquid nitrogen vapor phase
Images
Genes Expressed plasminogen activator
Cellular Products plasminogen activator
Virus Susceptibility Bovine viral diarrhea virus 1 , Bovine viral diarrhea virus 1 Human poliovirus 1 Human Coxsackievirus B4
Comments The serum used in the medium should be pre-tested for good quality. Some lots of horse serum do support satisfactory growth.

Adaptation to other types of sera generally results in slow growth with a prolonged lag in growth.
Complete Growth Medium Medium 199 (with 1.68 g/L bicarbonate), 99%; horse serum, 1%
Subculturing

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM  EDTA solution to remove all traces of serum, which contains trypsin inhibitor.
  3. 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 with 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. To remove trypsin-EDTA solution, transfer cell suspension to  centrifuge tube and spin atapproximately 125 x g for 5 to10 minutes.
  6. Discard supernatant and resuspend cells in fresh culture medium.  Add appropriate aliquots of cell suspension to new culture vessels. 
  7. Place culture vessels in incubators at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:10 is recommended

Medium Renewal: Every 2 to 3 days
Cryopreservation Culture medium, 95%; DMSO, 5%
Culture Conditions Temperature: 37°C
Name of Depositor RN Hull
References

American Association of Anatomists. Sixty-ninth annual session; Marquette University School of Medicine, Milwaukee, Wisconsin. Anat. Rec. 124: 490, 1956.

Hull RN, et al. Growth characteristics of monkey kidney cell strains LLC-MK1, LLC-MK2, and LLC-MK2(NCTC-3196) and their utility in virus research. J. Exp. Med. 115: 903-918, 1962. PubMed: 14449901

Hull RN, Tritch OJ. Characterization of cell strains by viral susceptibility. Natl. Cancer Inst. Monogr. 7: 161-172, 1962. PubMed: 14449902

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