B-3 cell line细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心

B-3 cell line细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心
Organism Homo sapiens, human Tissue eye; lens Cell Type transformed with an adenovirus 12-SV40 virus hybrid (Ad12-SV40) Product Format frozen Morphology epithelial Culture Properties adherent Biosafety Level 2
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 Ref Genes Expressed beta and gamma crystallin (Andley UP, Fleming TP. Immortalized epithelial cell lines. US Patent 5,643,782 dated Jul 1 1997) Cellular Products beta and gamma crystallin (Andley UP, Fleming TP. Immortalized epithelial cell lines. US Patent 5,643,782 dated Jul 1 1997) 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. 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. Remove and add additional Trypsin-EDTA solution to flask. Incubate at room temperature for 60 seconds or less. Immediately add complete growth medium and aspirate cells by gently pipetting. 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%
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