Roche FuGENE® Transfection Reagents）

Tested and found negative for ectromelia virus （mousepox）.

This line is also designated as ATCC CCL-92.1.ATCC CL-173 was deposited in 1974 without passage number information from the depositor. At the time of submission, ATCC prepared approximay 30 vials of seed stock at about 4 passages beyond the original depositor material （passage number: unknown +4）.

1. Remove and discard culture medium.

2. 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.

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 37C 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.

   The recommended inoculum is 2 to 3 X 10（3） cells/sq. cm. Subculture before cultures become 70 to 80% confluent or when cells reach 5 to 6 X10（4） viable cells/sq. cm.

6. Incubate cultures at 37C.

formerly distributed as:ATCC CCL-92.1

0.25% （w/v） Trypsin - 0.53 mM EDTA in Hank' BSS （w/o Ca++, Mg++）:ATCC 30-2101

Cell culture tested DMSO:ATCC 4-X

Recommended serum: ATCC 30-2030

3491: Green H. Triglyceride-accumulating clonal cell line. US Patent 4,003,789 dated Jan 18 1977

32373: Goodrum FD, et al. Adenovirus early region 4 34-kilodalton protein directs the nuclear localization of the early region 1B 55-kilodalton protein in primate cells. J. Virol. 70: 6323-6335, 1996. PubMed: 8709260