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This human cell line contained large numbers of tetra-, hexa-, and higher-ploid cells (2s populations). The modal (s) cell population which occurred in 32% of cells had the pseudodiploid karyotype, 46,XX,-3,-18,del(7) (q21.12;q22.3), ?t(3q?18q). The rate of 2s cells was 42%. Two marker chromosomes, del(7) (q21.12;q22.3) and ?t(3q?18q), were present in all s metaphases, but ?t(3q?18q) was absent in 2s cells. DMs were found in some s cells whereas they were seen in the majority of 2s cells. HSR chromosomes were not found. In s metaphases, both N3 and N18 had only single copy, and the X chromosome was paired.
Age:
63 years
Gender:
female
Ethnicity:
Caucasian
Comments:
This line was derived from a primary clear cell adenocarcinoma. The cells are globular with indistinct borders, have a high nucleus to cytoplasm ratio and exhibit both microvilli and desmosomes. They can be grown in soft agar.
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Temperature: 37.0°C
Subculturing:
Protocol:
Remove and discard culture medium.
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.
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.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:12 is recommended Medium Renewal: Every 2 to 3 days
Preservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Doubling Time:
35 hrs
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001 recommended serum:ATCC 30-2020
References:
22648: Williams RD, et al. In vitro cultivation of human renal cell cancer. I. Establishment of cells in culture. In Vitro 12: 623-627, 1976. PubMed: 1010528 22653: Williams RD, et al. In vitro cultivation of human renal cell cancer. II. Characterization of cell lines. In Vitro 14: 779-786, 1978. PubMed: 721102
1. Treat cells. For PC12 or NIH 3T3 cells a moderately confluent 60 mm plate is used. 2. Aspirate media. Rinse cells with PBS, aspirate.3. Lyse cells in 200 uL of SDS sample buffer (100 ℃). Immediately scrape the cells off the plate with a rubber
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