Permits and Restrictions |
View Permits View Restrictions |
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Organism | Homo sapiens, human |
Tissue | lung, bronchial |
Cell Type | epithelial |
Product Format | frozen |
Morphology | epithelial, packed cuboidal |
Culture Properties | adherent |
Biosafety Level |
2 [Cells immortalized by CDK4 and hTERT and contain SV40 promoter sequences]
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 | normal |
Age | 65 |
Gender | female |
Applications |
HBEC3-KT cells are normal human bronchial epithelial cells immortalized with CDK4 and hTERT. The immortalized HBEC3-KT cells do not form colonies in soft agar, nor do they form tumors in mice (RefRamirez RD, Sheridan S, Girard L, et al. Immortalization of human bronchial epithelial cells in the absence of viral oncoproteins. Cancer Res. 64(24):9027-9034, 2004. PubMed: 15604268). Multiple oncogenic changes (K-RAS(V12), p53 knockdown, mutant EGFRs) are not sufficient to confer a full malignant phenotype on HBEC3-KT. These additional genetic changes, commonly found in human lung cancer, progress the HBEC3-KT cells partially, but not completely, toward malignancy (RefSato M, et al. Multiple oncogenic changes (K-RAS(V12), p53 knockdown, mutant EGFRs, p16 bypass, telomerase) are not sufficient to confer a full malignant phenotype on human bronchial epithelial cells. Cancer Res. 66(4): 2116-2128, 2006. PubMed: 16489012). Extended lifespan (RefRamirez RD, Sheridan S, Girard L, et al. Immortalization of human bronchial epithelial cells in the absence of viral oncoproteins. Cancer Res. 64(24):9027-9034, 2004. PubMed: 15604268), multi-potent differentiation capacity in three-demensional models (RefDelgado O, et al. Multipotent capacity of immortalized human bronchial epithelial cells. PLoS ONE. 6(7): e22023, 2011. PubMed: 21760947), and drug-sensitivity tests (paclitaxel, carboplatin, pemetrexed, cisplatin, gemcitabine, etc.) have been reported for the HBEC3-KT cells (RefSato M, et al. Human lung epithelial cells progressed to malignancy through specific oncogenic manipulations. Mol. Cancer Res. 11(6): 638-650, 2013. PubMed: 23449933). |
Storage Conditions | liquid nitrogen vapor phase |
Karyotype | Cytogenetic analysis was performed on G-banded metaphase cells from the human cell line HBEC3-KT, and demonstrated mixed population of clones with near-diploid or tetraploid abnormal female karyotypes. Unbalanced translocations der(16)t(5;16)(q11.2;q24) and der(13)t(8;13)(q13.1;p11.1) as well as an extra copy of chromosome 20 with additional translocation +add(20)(q13.3) are observed in these clones. These aberrations result in partial trisomies of chromosome 5, 8 or 20 which have been found in a previous aCGH microarray analysis of HBEC3-KT (PMID: 15604268) |
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Images | |
Derivation |
The HBEC3-KT cell line was established by infecting primary human bronchial epithelial cell culture with human telomerase (hTERT) and mouse cyclin dependent kinase 4 (CDK4) expressing retrovirus constructs and selecting with Puromycin and G418 as described in PMID: 15604268 (RefRamirez RD, Sheridan S, Girard L, et al. Immortalization of human bronchial epithelial cells in the absence of viral oncoproteins. Cancer Res. 64(24):9027-9034, 2004. PubMed: 15604268). |
Clinical Data | female 65 years |
Antigen Expression | Positive for p63 (TP63) and Clara cell 10 (CC10) protein |
Complete Growth Medium | Airway Epithelial Cell Basal Medium (ATCC PCS-300-030) suplemented with Bronchial Epithelial Cell Growth Kit (ATCC PCS-300-040) |
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Subculturing |
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation solutions for culture vessels of other sizes. Subculture when the culture is about 70-80% confluent.
Note: Cells are sensitive to FBS solution. |
Cryopreservation | 80% complete growth media, 10% DMSO, 10% FBS |
Culture Conditions | Atmosphere: air, 95%; carbon dioxide (CO2), 5%Temperature: 37°C |
STR Profile | D5S818: 12 D13S317: 12, 13 D7S820: 8, 12 D16S539: 9, 12 vWA: 15, 17 Amelogenin: X TPOX: 11, 12 CSF1PO: 10, 11 TH01: 9.3 |
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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. |
Name of Depositor | John Minna, Shelley Sheridan |
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References |
Ramirez RD, Sheridan S, Girard L, et al. Immortalization of human bronchial epithelial cells in the absence of viral oncoproteins. Cancer Res. 64(24):9027-9034, 2004. PubMed: 15604268 Delgado O, et al. Multipotent capacity of immortalized human bronchial epithelial cells. PLoS ONE. 6(7): e22023, 2011. PubMed: 21760947 Sato M, et al. Human lung epithelial cells progressed to malignancy through specific oncogenic manipulations. Mol. Cancer Res. 11(6): 638-650, 2013. PubMed: 23449933 Sato M, et al. Multiple oncogenic changes (K-RAS(V12), p53 knockdown, mutant EGFRs, p16 bypass, telomerase) are not sufficient to confer a full malignant phenotype on human bronchial epithelial cells. Cancer Res. 66(4): 2116-2128, 2006. PubMed: 16489012 Vaughan MB, et al. A three-dimensional model of differentiation of immortalized human bronchial epithelial cells. Differentiation. 74(4):141-148, 2006. PubMed: 16683984 |