CP-D (CP-18821)細胞

運輸方式： 凍存運輸

ATCC Number： CRL-4030?

相關(guān)**： Barrett食管

年限： *****

細胞類型： 其他細胞類型

細胞形態(tài)： 上皮樣

是否是腫瘤細胞： 0

物種來源： 人

組織來源： epithelium

器官來源： 食道

生長狀態(tài)： 貼壁生長

數(shù)量： 大量

Designations: CP-D (CP-18821)

Depositors: B. Reid

CP-D (CP-18821)細胞Biosafety Level: 2

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: epithelial-like

Source: Organ: esophagus

Tissue: epithelium

Cell Type: high-grade dysplasia

Disease: Barrett's esophagus

Immortalization method: hTERT expression

Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

Restrictions: CP-D (CP-18821)細胞This material requires that the Addendum for Commercial and For-Profit Organizations or the Addendum for Noncommercial and Academic Organizations be signed and returned to ATCC before shipment. The price listed above is for noncommercial and academic organizations only. Commercial and for-profit organizations should call for pricing.

Isolation: Isolation date: April 1995

Applications: In addition, it has been verified that no gross changes are observed in karyotype and morphology during the first 10 population doublings.

As part of our quality control, we have tested this cell line for its ability to grow for a minimum of 15 population doublings after recovery from cryopreservation.

The Barrett's esophagus cell line, CP-D (also identified as CP-18821) was derived from an endoscopic biopsy specimen obtained from a region of high-grade dysplasia.

Genetic instability studies using flow cytometry and FISH reveal the retention of elevated tetraploidy (G2/tetraploidy) in the hTERT-immortalized cells, similar to the non-transduced parental cells.

Morphologically, the cell line is similar to early passage cultures exhibiting smaller cells with large nucleus to cytoplasm ratio.

Antigen Expression: positive for epithelial marker pan-cytokeratin (immunocytochemistry)(verified at ATCC )

negative for gastric mucin (CLH2) (immunocytochemistry)(verified at ATCC )

DNA Profile (STR): CSF1PO: 12

D13S317: 12

D16S539: 10, 13

D5S818: 9, 12

D7S820: 10, 11

THO1: 9, 9.3

TPOX: 8

vWA: 16

Amelogenin: CP-D (CP-18821)細胞X (Note: LOH of Y)

Cytogenetic Analysis: This is a hypotetraploid human cell line with the following derivative chromosomes consistently present at several different passages: add(2)(q13), der(3)t(3;8)(p10;q10), ider(7)(q10)dup(q31), der(12)t(12;13)(p10;q10), der(14)t(14;15)(q10;q10), der(15)t(15;22)(q10;q10), add(22)(q13)x2. In addition, there were consistent losses of one copy of chromosomes X, 10, 13, 14, 15, 19 and 20. Other less consistent structural aberrations were observed in some of the examined cells.

Age: *****

Gender: male

Comments: The Barrett's esophagus cell line, CP-D (also identified as CP-18821) was derived from an endoscopic biopsy specimen obtained from a region of high-grade dysplasia. The cells were immortalized by transduction with a retroviral expression vector, pLXSN-hTERT, to create an immortalized cell line. [16173160]

Terminal restriction fragment lengths (TRF) analyses show the cells have increased telomerase activity and extended telomeres of about 12 kb. Morphologically, the cell line is similar to early passage cultures exhibiting smaller cells with large nucleus to cytoplasm ratio. Genetic instability studies using flow cytometry and FISH reveal the retention of elevated tetraploidy (G2/tetraploidy) in the hTERT-immortalized cells, similar to the non-transduced parental cells. [16173161]

This well-characterized pre-malignant culture represents a unique tool for studying esophageal cancer progression. [16173160]

As part of our quality control, we have tested this cell line for its ability to grow for a minimum of 15 population doublings after recovery from cryopreservation. In addition, it has been verified that no gross changes are observed in karyotype and morphology during the first 10 population doublings.

Propagation: CP-D (CP-18821)細胞ATCC complete growth medium: The base medium for this cell line is MCDB-153. To make the complete growth medium, add the following components to the base medium:

0.4 ?g/ml hydrocortisone

20 ng/ml recombinant human EGF (Epidermal Growth Factor)

1 nM cholera toxin

20 mg/L adenine

140 ?g/ml BPE (Bovine Pituitary Extract)

0.1% ITS [Insulin-Transferrin-Sodium Selenite Supplement(Sigma, I1884)]

4 mM glutamine

fetal bovine serum to a final concentration of 5%

Temperature: 37.0°C

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

Subculturing: Protocol: Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium.

Add 2.0 to 3.0 ml of 0.25% (w/v) Trypsin - 0.53 mM 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 37.0°C to facilitate dispersal.

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

Transfer cell suspension to a centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes. Discard supernatant.

Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1 X 104 to 2 X 104 viable cells/cm2 is recommended.

Incubate cultures at 37.0°C. CP-D (CP-18821)細胞Subculture when cells reach a concentration between 7 X 104 and 1 X 105 cells/cm2.

Subcultivation ratio: A subcultivation ratio of 1:2 to 1:5 is recommended.

Medium renewal: every 3 to 4 days

Preservation: Freeze medium: RPMI-1640 Medium, 80%; fetal bovine serum, 10%; DMSO, 10%

Storage temperature: liquid nitrogen vapor phase

Doubling Time: approximately 39 hours

Related Products: Recommended serum: ATCC 30-2020

0.25% (w/v) Trypsin - 0.53mM EDTA in Hank's BSS (w/o Ca++, Mg ++): ATCC 30-2101

Cell culture tested DMSO: ATCC 4-X

Recommended base medium for freezing (without the additional serum described under Freeze Medium): ATCC 30-2001

References: 16173160: Palanca-Wessels MC, et al. Genetic Analysis of Long-term Barrett's Esophagus Epithelial Cultures Exhibiting Cytogenetic and Ploidy Abnormalities. Gastroentrology 114:114-295, 1998. PubMed: 9453489

16173161: Palanca-Wessels MC, et al. Extended lifespan of Barrett's esophagus epithelium transduced with the human telomerase catalytic subunit: a useful in vitro model. Carcinogenesis 24(7): 1183-1190, 2003. PubMed: 12807723

16173615: Barrett MT, et al. Molecular Phenotype of Spontaneously Arising 4N (G2-Tetraploid) Intermediates of Neoplastic Progression in Barrett's Esophagus. Cancer Res. 63: 4211-4217, 2003. PubMed: 12874028

16173616: Maley CC, et al. Genetic clonal diversity predicts progression to esophageal adenocarcinoma. Nat. Genet. 38(4): 468-473, 2006. PubMed: 16565718