HCT 116細(xì)胞

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

數(shù)量： 大量

ATCC Number： CCL-247?

相關(guān)**： 大腸癌

運(yùn)輸方式： 凍存運(yùn)輸

細(xì)胞形態(tài)： 上皮樣

年限： *****

是否是腫瘤細(xì)胞： 1

物種來(lái)源： 人

器官來(lái)源： 結(jié)腸

Designations: HCT 116

HCT 116細(xì)胞Depositors: MG Brattain

Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: epithelial

Source: Organ: colon

Disease: colorectal carcinoma

Cellular Products: carcinoembryonic antigen (CEA) 1 ng per 10 exp6 cells per 10 days; keratin

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Applications: transfection host

Tumorigenic: Yes

DNA Profile (STR): HCT 116細(xì)胞Amelogenin: X,Y

CSF1PO: 7,10

D13S317: 10,12

D16S539: 11,13

D5S818: 10,11

D7S820: 11,12

THO1: 8,9

TPOX: 8,9

vWA: 17,22

Cytogenetic Analysis: The stemline chromosome number is near diploid with the modal number at 45 (62%) and polyploids occurring at 6.8%. The markers 10q+ and t(?8p;18q) are present in all metaphases and t(9q;?16p-), in 80% of the cells karyotyped. N16 is monosomic in the presence of, but disomic in the absence of t(9q;?16p-). N10 and N18 are monosomic and other chromosomes from those mentioned above are disomic. Q-band observations revealed the presence of the Y chromosome, but not in all cells (50% of cells lacked the Y in G-band karyotypes).

Isoenzymes: AK-1, 1

ES-D, 1-2

G6PD, B

GLO-I, 1

PGM1, 1

PGM3, 1

HCT 116細(xì)胞Age: *****

Gender: male

Comments: The cells are positive for keratin by immunoperoxidase staining.

HCT 116 cells are positive for transforming growth factor beta 1 (TGF beta 1) and beta 2 (TGF beta 2) expression.

This line has a mutation in codon 13 of the ras protooncogene, and can be used as a positive control for PCR assays of mutation in this codon.

Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated McCoy's 5a Medium Modified, Catalog No. 30-2007. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.

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

Temperature: 37.0°C

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

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 which 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: HCT 116細(xì)胞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 1:3 to 1:8 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

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

recommended serum:ATCC 30-2020

Trypsin EDTA Solution: ATCC 30-2101

Cell culture tested DMSO: ATCC 4-X

References: 22794: 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

23040: Brattain MG, et al. Heterogeneity of malignant cells from a human colonic carcinoma. Cancer Res. 41: 1751-1756, 1981. PubMed: 7214343

23125: Sun L, et al. HCT 116細(xì)胞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

25093: Santoro IM, Groden J. Alternative splicing of the APC gene and its association with terminal differentiation. Cancer Res. 57: 488-494, 1997. PubMed: 9012479

26071: 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

32266: 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

32288: 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

32794: 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

32910: 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