亚洲.无码.制服.日韩.中I人妻无码久久精品人妻成人I制片厂91I国产真实91东北熟妇HDXXXI五月AVI四虎精品性爱I国产亚州avI日产精品久久久久I人人爱夜夜操

SW620 was initiated by A. Leibovitz, et al., from a lymph node in the same manner as was the primary adenocarcinoma from which SW480 was derived the previous year.It was isolated from the tissue of a 51-year-old Caucasian male (blood group A, Rh+) as was SW480 (ATCC CCL-228).

The established cell line consists mainly of individual small spherical and bipolar cells lacking microvilli.

The cells are negative for expression of CSAp (CSAp-) and colon antigen 3, negative.

The line was derived from a metastasis of the same tumor from which the SW480 (ATCC CCL-228) was derived.

The line is positive for expression of c-myc, K-ras, H-ras, N-ras, Myb, sis and fos oncogenes.

N-myc oncogene expression was not detected.

(Note: The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation)

Volumes used in this protocol are for 75 cm² flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning^® T-75 flasks (catalog #430641) are recommended for subculturing this product.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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 2.0 to 3.0 mL of complete growth medium and aspirate cells by gently pipetting
5. Resuspend the cell pellet in fresh growth medium. 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 1:10 is recommended
Medium Renewal: 2 to 3 times per week

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Fogh J, et al. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J. Natl. Cancer Inst. 58: 209-214, 1977. PubMed: 833871

Fogh J, et al. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J. Natl. Cancer Inst. 59: 221-226, 1977. PubMed: 327080

Goodfellow M, et al. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J. Natl. Cancer Inst. 59: 221-226, 1977. PubMed: 77210034

Lelbovitz A, et al. Detection and analysis of a glucose 6-phosphate dehydrogenase phenotype B cell line contamination. J. Natl. Cancer Inst. 63: 635-645, 1979. PubMed: 288927

Trainer DL, et al. Biological characterization and oncogene expression in human colorectal carcinoma cell lines. Int. J. Cancer 41: 287-296, 1988. PubMed: 3338874

Leibovitz A, et al. Classification of human colorectal adenocarcinoma cell lines. Cancer Res. 36: 4562-4569, 1976. PubMed: 1000501

Rodrigues NR, et al. p53 mutations in colorectal cancer. Proc. Natl. Acad. Sci. USA 87: 7555-7559, 1990. PubMed: 1699228

Witty JP, et al. Modulation of matrilysin levels in colon carcinoma cell lines affects tumorigenicity in vivo. Cancer Res. 54: 4805-4812, 1994. PubMed: 8062282

Gagos S, et al. Chromosomal markers associated with metastasis in two colon cancer cell lines established from the same patient. Anticancer Res. 15: 369-378, 1995. PubMed: 7763008

Melcher R, et al. Spectral karyotyping of the human colon cancer cell lines SW480 and SW620. Cytogenet. Cell Genet. 88: 145-152, 2000. PubMed: 10773689

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.