B-3 (ATCC® CRL-11421)

Organism: Homo sapiens, human  /  Tissue: eye; lens  / 

Organism Homo sapiens, human
Tissue
eye; lens
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain Adenovirus-SV-40 viral DNA sequences]
Age infant less than 1 year
Storage Conditions liquid nitrogen vapor phase
Derivation
The cell line was derived from a human lens obtained within 24 hrs from 5-12 month old patient who underwent treatment for retinopathy of prematurity. At 60% confluence, cells at passage 3 were infected with Ad12-SV40.
Clinical Data
The cell line was derived from a human lens obtained within 24 hrs from 5-12 month old patient who underwent treatment for retinopathy of prematurity.
Antigen Expression
SV40 T antigen
Genes Expressed
beta and gamma crystallin
Cellular Products
beta and gamma crystallin
Comments
The cells synthesize beta and gamma crystallins as monitored by immunoblot assay.

The cells can be maintained in culture for over 76 population doublings with no decrease in proliferative capability.

Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 20%.
Subculturing
These cells are very sensitive to trypsin-EDTA.
  1. Quickly rinse (less than 30 seconds) the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum, which contains trypsin inhibitor.
  2. Remove and add additional Trypsin-EDTA solution to flask.
  3. Incubate at room temperature for 60 seconds or less.
  4. Immediately add complete growth medium and aspirate cells by gently pipetting.
  5. Dispense into new culture flasks.
    Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended
    Cryopreservation
    Freeze Medium: Complete growth medium supplemented with 5% (v/v) DMSO
    Storage Temperature: Liquid nitrogen vapor phase
    Culture Conditions
    Temperature: 37°C
    Atmosphere: Air, 95%; carbon dioxide (CO2), 5%
    Name of Depositor T Fleming, U Andley
    Passage History
    At 60% confluence, cells at passage 3 were infected with Ad12-SV40.
    References

    Andley UP, Fleming TP. Immortalized epithelial cell lines. US Patent 5,643,782 dated Jul 1 1997

    Andley UP, et al. Propagation and immortalization of human lens epithelial cells in culture. Invest. Ophthalmol. Vis. Sci. 35: 3094-3102, 1994. PubMed: 8206728

    Fleming TP, et al. Expression of growth control and differentiation genes in human lens epithelial cells with extended life span. Invest. Ophthalmol. Vis. Sci. 39: 1387-1398, 1998. PubMed: 9660487

    Basic Documentation
    References

    Andley UP, Fleming TP. Immortalized epithelial cell lines. US Patent 5,643,782 dated Jul 1 1997

    Andley UP, et al. Propagation and immortalization of human lens epithelial cells in culture. Invest. Ophthalmol. Vis. Sci. 35: 3094-3102, 1994. PubMed: 8206728

    Fleming TP, et al. Expression of growth control and differentiation genes in human lens epithelial cells with extended life span. Invest. Ophthalmol. Vis. Sci. 39: 1387-1398, 1998. PubMed: 9660487