CP-D; Esophagus Epithelium; Human (Homo sapiens) Item ID: ATCCRL4030
| Permits and Restrictions |
View Permits View Restrictions |
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| Organism | Homo sapiens, human |
| Tissue | esophagus; epithelium |
| Cell Type | high-grade dysplasia |
| Product Format | frozen |
| Morphology | epithelial-like |
| Culture Properties | adherent |
| Biosafety Level |
2
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease | Cancer, Barrett's esophagus |
| Age | adult |
| Gender | male |
| Applications | This well-characterized pre-malignant culture represents a unique tool for studying esophageal cancer progression. |
| Storage Conditions | liquid nitrogen vapor phase |
| Karyotype | 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. |
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| Images | |
| Derivation | 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. |
| Clinical Data | male |
| Antigen Expression | positive for epithelial marker pan-cytokeratin (immunocytochemistry)(verified at ATCC) negative for gastric mucin (CLH2) (immunocytochemistry)(verified at ATCC) |
| Genes Expressed | positive for epithelial marker pan-cytokeratin (immunocytochemistry)(verified at ATCC),negative for gastric mucin (CLH2) (immunocytochemistry)(verified at ATCC) |
| Comments |
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. 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. |
| 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:
Note: To prepare Cholera toxin (Stock 100 µg/mL) : 0.5 mg Cholera toxin (Sigma C8052) + 5 mL dH20. Aliquot into microcentrifuge tubes. Add 84 µL of this 100 µg/mL stock solution to 1L of MCDB-153 base medium. |
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| Subculturing |
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
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| Cryopreservation | Freeze medium: RPMI-1640 Medium, 80%; fetal bovine serum, 10%; DMSO, 10% Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions | Temperature: 37.0°C Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| STR Profile | CSF1PO: 12 D13S317: 12 D16S539: 10, 13 D5S818: 9, 12 D7S820: 10, 11 THO1: 9, 9.3 TPOX: 8 vWA: 16 Amelogenin: X (Note: LOH of Y) |
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| Population Doubling Level (PDL) | 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. We have also compared its karyotype, telomerase expression level, growth rate, morphology and tissue-specific markers when first recovered from cryopreservation with that of cells at 10+ population doublings to ensure that there is no change in these parameters and that the cells are capable of extended proliferation. |
| Population Doubling Time | approximately 39 hours |
| Name of Depositor | B. Reid |
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| Passage History | 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. |
| Year of Origin | April 1995 |
| References |
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 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 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 Maley CC, et al. Genetic clonal diversity predicts progression to esophageal adenocarcinoma. Nat. Genet. 38(4): 468-473, 2006. PubMed: 16565718 |