In Vitro Technologies

Permits and Restrictions	View Permits
Organism	Macaca mulatta, monkey, rhesus
Tissue	lung/bronchus
Cell Type	epithelial
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	2 to 3 years
Storage Conditions	liquid nitrogen vapor phase

Karyotype	The stemline modal chromosome number is 45 occurring at 68%, and polyploidy at 1.6%. The only marker chromosome 14 pt is found in the majority (60%) of cells. N3 is tetrasomic and N19 is trisomic. There are two copies of X chromosomes, but no Y-like chromosome. The cell line appears to be of female origin. Note: Cytogenetic information is based on initial seed stock at ATCC. Cytogenetic instability has been reported in the literature for some cell lines.
Derivation	This epithelial cell line was derived from the normal bronchial tissue of a 2- to 3-year-old rhesus monkey. The line was initiated in May, 1978 from tissue explants using glass chips.
Receptor Expression	epidermal growth factor (EGF), expressed
Genes Expressed	keratin
Comments	Cells contain PAS-positive cytoplasmic inclusions. The cells are positive for keratin by immunoperoxidase staining.

Complete Growth Medium	Ham's F-12K Medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and supplemented with 30ng/ml murine epidermal growth factor [BD Bioscience (BD354001) or Sigma (E-4127)] and 10% fetal bovine serum
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. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that 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. Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium Renewal: 2 to 3 times per week
Cryopreservation	Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions	Atmosphere: air, 95%; carbon dioxide (CO₂), 5% Temperature: 37°C Growth Conditions: The line requires murine epidermal growth factor (EGF) for proliferation, which may be aquired from BD Bioscience (BD354001) or Sigma (E-4127).

Name of Depositor	JL Caputo
Year of Origin	May, 1978
References	Caputo JL, Hay RJ, Williams CD. The isolation and properties of an epithelial cell strain from rhesus monkey bronchus. In Vitro 15: 222-223, 1979. Rubin JS, et al. Purification and characterization of a newly identified growth factor specific for epithelial cells. Proc. Natl. Acad. Sci. USA 86: 802-806, 1989. PubMed: 2915979

Name of Depositor

JL Caputo

Year of Origin

May, 1978

References

Caputo JL, Hay RJ, Williams CD. The isolation and properties of an epithelial cell strain from rhesus monkey bronchus. In Vitro 15: 222-223, 1979.

Rubin JS, et al. Purification and characterization of a newly identified growth factor specific for epithelial cells. Proc. Natl. Acad. Sci. USA 86: 802-806, 1989. PubMed: 2915979

E: care@invitro.com.au
P: 1300 552 003

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4MBr-5; Lung; Rhesus Monkey (Macaca mulatta) Item ID: ATCCCL208

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