BE CP-A cell line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

BE CP-A cell line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心
BE CP-A cell line细胞株
Organism Homo sapiens, human
Tissue Esophagus; epithelium
Cell Type Epithelial cells immortalized with hTERT
Product Format frozen
Morphology Epithelial-like
Culture Properties Adherent
Biosafety Level 2 [Cells contain SV40 viral DNA sequences]
Disease Non-dysplastic metaplasia
Age Adult
Gender Male
Applications This well-characterized pre-malignant culture represents a unique opportunity to study factors that are important in both cancer progression and hTERT immortalization.
Karyotype This is a near-diploid cell line of male origin in which 2 sub-clones make up the majority of the cell population. One clone containing i(8)(q10) and trisomy 20 and the other containing der(1)t(1;18)(q10;q10), i(8)(q10), der(13)t(13;22)(q10;q10) and trisomy 20. The remaining population is generally made up of cells with non-clonal aberrations that were derived from the two major clones. Also, the non-clonal cell population may increase at high passages.
Images
Derivation The Barrett's esophagus cell line, CP-A (also identified as KR-42421 or QhTERT), was derived from an endoscopic biopsy specimen obtained from a region of non-dysplastic metaplasia and transduced with the retroviral expression vector, pLXSN-hTERT, to create an immortalized cell line.
Antigen Expression Positive for epithelial marker pan-cytokeratin (immunocytochemistry) (verified at ATCC)
Negative for gastric mucin (CHL2) (immunocytochemistry) (verified at ATCC)
Comments This cell line shows increased telomerase activity and long telomeres of about 12 kb as assessed by terminal restriction fragment lengths (TRF) analysis. Morphologically, the cells are similar to early passage cultures: smaller cells with a larger 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.
STR Profile Amelogenin: X,Y
CSF1PO:12
D13S317:12,14
D16S539:12,13
D5S818:11,12
D7S820:7,8
THO1: 7,9.3
TPOX: 8,9
vWA: 17
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.
Year of Origin August 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


Complete Growth Medium The base medium for this cell line is MCDB-153. For complete growth medium, add the following components to the base medium:
• 0.4 µg/ml hydrocortisone
• 20 ng/ml recombinant human epidermal growth factor
• 8.4 µg/L cholera toxin
• 20 mg/L adenine
• 140 µg/ml bovine pituitary extract
• 1x ITS Supplement (Sigma; I1884)
• 4 mM glutamine
• fetal bovine serum to a final concentration of 5%
To prepare Cholera toxin (Stock 100 µg/mL): 0.5 mg Cholera toxin (Sigma C8052) + 5 mL dH20. Add 84 µL of this 100 µg/mL stock solution to 1L of MCDB-153 base medium. Do not filter the complete medium.

Cryopreservation Freeze Medium: RPMI-1640 Medium, 80%; fetal bovine serum, 10%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%

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