The cells are distributed for research purposes only. The Memorial Sloan-Kettering Cancer Center releases the line subject to the following: 1.) The cells or their products must not be distributed to third parties. Commercial interests are the exclusive property of Memorial Sloan-Kettering Cancer Center. 2.) Any proposed commercial use of these cells must first be negotiated with The Director, Office of Industrial Affairs, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021; phone (212) 639-6181; FAX (212) 717-3439.

modal number = 46; range = 40 to 53.
This is a pseudodiploid human cell line with the modal chromosome number of 46, occurring in 56% of cells. The rate of polyploidy was 1.9%. There were no constitutive marker chromosomes. The N2 was trisomic. There was a high incidence of Y chromosome losses, which contributed to the high number of cells with 46 chromosomes.

34 years

Caucasian

Protocol:

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 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: A subcultivation ratio of 1:2 to 1:3 is recommended
Interval: Subculture every 6 to 8 days.

Medium renewal: 2 to 3 times per week

Recommended medium (without the additional supplements or serum described under ATCC Medium): ATCC 30-2007
recommended serum: ATCC 30-2020