宁波泰斯拓生物

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12MBr6

货号 TS213066
中文名称 null
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产品名称: 12MBr6
商品货号: TS213066
Organism: Cercopithecus aethiops
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
Gender: male
Applications:
Epidermal growth factor (EGF) was used to initiate and maintain growth.
Post-freeze longevity studies indicate that EGF is an essential component of the culture medium for successful long-term propagation of the cells.
12MBr6 is an epithelial cell line derived from secondary bronchial tissue of a 2- to 3-year-old normal African green monkey.
Storage Conditions: liquid nitrogen vapor phase
Karyotype: modal number = 62; range = 43 to 64.
The modal chromosome number was found in 78% of the cell population. The rate of polyploidy was 2.4%. Three consistent chromosome markers were identified: M1: del(1)9 ?; M2 del(13) and M3. Chromosome #20 is trisomic and the X chromosome is found in the disomic condition.
Derivation:
12MBr6 is an epithelial cell line derived from secondary bronchial tissue of a 2- to 3-year-old normal African green monkey. This line was initiated in June, 1979 at the American Type Culture Collection from tissue explants using penicylinders and a trypsin-versene solution for cell dispersal. These cells contain PAS-positive cytoplasmic inclusions. Epidermal growth factor (EGF) was used to initiate and maintain growth. Post-freeze longevity studies indicate that EGF is an essential component of the culture medium for successful long-term propagation of the cells. Progeny from the 12MBr6 were found to undergo at least 23 additional doublings beyond the frozen state.
Clinical Data:
male
Receptor Expression:
epidermal growth factor (EGF)
Comments:
12MBr6 is an epithelial cell line derived from secondary bronchial tissue of a 2- to 3-year-old normal African green monkey. This line was initiated in June, 1979 at the American Type Culture Collection from tissue explants using penicylinders and a trypsin-versene solution for cell dispersal. These cells contain PAS-positive cytoplasmic inclusions. Epidermal growth factor (EGF) was used to initiate and maintain growth. Post-freeze longevity studies indicate that EGF is an essential component of the culture medium for successful long-term propagation of the cells. Progeny from the 12MBr6 were found to undergo at least 23 additional doublings beyond the frozen state.
Complete Growth Medium: Hams 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.
  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 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:2 to 1:4
Medium Renewal: Every 2 to 3 days
Note: The line requires epidermal growth factor as a supplement to the medium.

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation:

Complete growth medium described above supplemented with 5% (v/v) DMSO.xa0 Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions:
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor: JL Caputo
Deposited As: Cercopithecus aethiops
Year of Origin: July, 1979
References:

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.