293AD Cell Line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

293AD Cell Line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心 The 293 Cell Line is a permanent line established from primary embryonic human kidney transformed
with human adenovirus type 5 DNA. The genes encoded by the E1 region of adenovirus (E1a and E1b)
are expressed in these cells and participate in transactivation of viral promoters, allowing these cells to
produce high levels of protein. E1 also complements the E1-deletion in recombinant adenoviral vectors,
allowing viral replication.
Background
293AD is derived from the parental 293 cell line, but specifically selected for adenovirus applications. It
offers several advantages over the regular 293 cells:
• Flattened morphology
• Firm attachment to culture plate, ideal for amplification and tittering of adenovirus
• Larger cell surface area resulting higher transfection and better yield of recombinant
adenovirus.
.
Quality Control
1. Culture Medium: D-MEM (high glucose), 10% fetal bovine serum (FBS), 0.1 mM MEM NonEssential
Amino Acids (NEAA), 2 mM L-glutamine, 1% Pen-Strep (optional)
Medium
2. Freeze Medium: 90% complete medium, 10% DMSO
I. Establishing 293AD Cultures from Frozen Cells
Methods
1. Place 10 mL of complete DMEM growth medium in a 50-mL conical tube. Thaw the frozen
cryovial of cells within 1–2 minutes by gentle agitation in a 37°C water bath. Decontaminate the
cryovial by wiping the surface of the vial with 70% (v/v) ethanol.
2. Transfer the thawed cell suspension to the conical tube containing 10 ml of growth medium.
3. Collect the cells by centrifugation at 1000 rpm for 5 minutes at room temperature. Remove the
growth medium by aspiration.
4. Resuspend the cells in the conical tube in 15 mL of fresh growth medium by gently pipetting up
and down.
5. Transfer the 15 mL of cell suspension to a T-75 tissue culture flask. Place the cells in a 37°C
incubator at 5% CO2.
6. Monitor cell density daily. Cells should be passaged when the culture reaches 95% confluence.
BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
电话：+86-010-53513060
网址：www.biovector.net [Supplier来源] http://www.biovector.net