GSB5005

HCT 116

Organism: Homo sapiens, human/ Tissue: colon / Disease: colorectal carcinoma

 

Catalog number

Unit size

List price(CNY)

1

GSB5005

-

1000

General information                                                                      

Product Format: Culturing in 25cm2 Flask

Morphology: epithelial

Culture Properties: adherent

Biosafety Level: 1

Gender: male

Applications:

This cell line is suitable as a transfection host. This line has a mutation in codon 13 of the ras proto-oncogene, and can be used as a positive control for PCR assays of mutation in this codon.

Characteristics

Karyotype:

The stemline chromosome number is near diploid with the modal number at 45 (62%) and polyploids occurring at 6.8%.

Tumorigenic:Yes

Images:

 

Culture method

Complete Growth Medium

The base medium for this cell line is McCoy's 5a Medium Modified. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.

Subculturing:

Volumes are given for a 75 cm2 flask. Corning® T-75 flasks are recommended for subculturing this product. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.

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 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 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 1:3 to 1:8 is recommended

Medium Renewal:  Every 2 to 3 days

Cryopreservation

Freeze medium: Complete growth medium 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

Culture Conditions

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37°C

Growth Conditions: Growth and plating efficiency are enhanced by using a feeder layer of murine fibroblasts.

References

Schroy PC, et al. Detection of p21ras mutations in colorectal adenomas and carcinomas by enzyme-linked immunosorbent assay. Cancer 76: 201-209, 1995. PubMed: 8625092

Brattain MG, et al. Heterogeneity of malignant cells from a human colonic carcinoma. Cancer Res. 41: 1751-1756, 1981. PubMed: 7214343

Sun L, et al. Autocrine transforming growth factor-beta 1 and beta 2 expression is increased by cell crowding and quiescence in colon carcinoma cells. Exp. Cell Res. 214: 215-224, 1994. PubMed: 8082724

Santoro IM, Groden J. Alternative splicing of the APC gene and its association with terminal differentiation. Cancer Res. 57: 488-494, 1997. PubMed: 9012479

Brattain MG, et al. Enhancement of growth of human colon tumor cell lines by feeder layers of murine fibroblasts. J. Natl. Cancer Inst. 69: 767-771, 1982. PubMed: 6956756

Bender CM, et al. Inhibition of DNA methylation by 5-Aza-2'-deoxycytidine suppresses the growth of human tumor cell lines. Cancer Res. 58: 95-101, 1998. PubMed: 9426064

Landers JE, et al. Translational enhancement of mdm2 oncogene expression in human tumor cells containing a stabilized wild-type p53 protein. Cancer Res. 57: 3562-3568, 1997. PubMed: 9270029

Kutchera W, et al. Protaglandin H synthase 2 is expressed abnormally in human colon cancer: evidence for a transcriptional effect. Proc. Natl. Acad. Sci. USA 93: 4816-4820, 1996. PubMed: 8643486

Wang R, et al. Cellular adherence elicits ligand-independent activation of the Met cell-surface receptor. Proc. Natl. Acad. Sci. USA 93: 8425-8430, 1996. PubMed: 8710887