產(chǎn)品名稱 |
4MBr-5 |
商品貨號(hào) |
B163772 |
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
Note: The line requires murine epidermal growth factor (EGF) for proliferation, which may be aquired from BD Bioscience (BD354001) or Sigma (E-4127). |
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 |
Name of Depositor |
JL Caputo |
Deposited As |
Macaca mulatta |
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
|