亞膜漢遜酵母

注意事項

1、 微生物菌種應保藏于低溫、清潔和干燥的地方，室溫放置時間過長會導致菌種衰退；

2、 菌種操作應在無菌條件下進行，防止雜菌污染；

3、 斜面菌種保藏時間通常為3-6個月，應根據菌種狀況及時轉接；凍干菌種保藏時間通常為2-15年；

亞膜漢遜酵母

ATCC® Number:  CL-188™

Designations:  LS 174T

Depositors:   Northwestern University

Biosafety Level: 1

Shipped:  frozen

Medium & Serum:  See Propagation

Growth Properties: adherent

Organism: Homo sapiens (human)

Morphology: epithelial

Source: Organ: colon

Tumor Stage: Dukes' type B

Disease: colorectal adenocarcinoma

Cellular Products: carcinoembryonic antigen (CEA) (The production of CEA in the ATCC seed stock was 1944 ng per 10(6) cells in 10 days.)

interleukin 10 (IL-10) [22511]

interleukin 6 (IL-6)

mucin

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Tumorigenic: Yes

Reverse Transcript: negative

Oncogene: myc +; myb + ; ras +; fos +; p53 +; sis -; abl -; ros -; src -

Antigen Expression: serologically defined colon cancer antigen 3; Homo sapiens, expressed

HLA A2, B13, B50; Blood type O

Cytogenetic Analysis: 45,X; one X chromosome missing; no other chromosomal aberrations

Isoenzymes:  ADA, 1

ES-D, 1

G6PD, B

PEP-D, 1

PGD, A

PGM1, 1

PGM3, 2

Age:  58 years

Gender:  female

Ethnicity:  Caucasian

Comments: LS 174T is a variant of LS 180 (ATCC CL-187) that has been maintained by using trypsin in the subculture protocol. It is more easily subc*ted than that parent line and, like LS 180, it is reported to produce large amounts of carcinoembryonic antigen (CEA).

Electron microscopic studies revealed abundant microvilli and intracytoplasmic mucin vacuoles [Pubmed ID: 1262041].

They are negative for p53 antigen expression, but positive for mRNA expression.

LS 174T cells stain positively for cytokeratins.

The line is positive for expression of c-myc, N-myc, H-ras, N-ras.

Myb, and fos oncogenes.

K-ras and sis oncogene expression were not detected.

Propagation:  ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. 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

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.

Subc*tion Ratio: A subc*tion ratio of 1:2 to 1:4 is recommended

Medium Renewal: 2 to 3 times per week

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 30-2003

recommended serum:ATCC 30-2020

derived from same individual:ATCC CL-187

References: 2148: Tom BH, et al. Human colonic adenocarcinoma cells. I. Establishment and description of a new line. In Vitro 12: 180-191, 1976. PubMed: 1262041

3524: Tom BH, et al. Process of producing carcinoembryonic antigen. US Patent 4,228,236 dated Oct 14 1980

22147: Chen TR, et al. Intercellular karyotypic similarity in near-diploid cell lines of human tumor origins. Cancer Genet. Cytogenet. 10: 351-362, 1983. PubMed: 6652615

22511: Gastl GA, et al. Interleukin-10 production by human carcinoma cell lines and its relationship to interleukin-6 expression. Int. J. Cancer 55: 96-101, 1993. PubMed: 8344757

22861: Trainer DL, et al. Biological characterization and oncogene expression in human colorectal carcinoma cell lines. Int. J. Cancer 41: 287-296, 1988. PubMed: 3338874

32265: Tsao H, et al. Novel mutations in the p16/CDKN2A binding region of the Cyclin-dependent Kinase-4 gene. Cancer Res. 58: 109-113, 1998. PubMed: 9426066

32377: Shibata D, et al. Genomic instability in repeated sequences is an early somatic event in colorectal tumorigenesis that persists after transformation. Nat. Genet. 6: 273-281, 1994. PubMed: 8012390

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