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HBEC3-KT

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編號(hào):B205492

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產(chǎn)品名稱	HBEC3-KT
商品貨號(hào)	B205492
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)
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)
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. Remove and discard spent medium. Briefly rinse the cells with Dulbecco's Phosphate Buffered Saline (DPBS, ATCC 30-2200), 1 mL / 25 cm² and discard rinse solution. Add Trypsin-EDTA, at 1 mL / 25 cm², for Primary Cells (ATCC PCS-999-003) to the flask. Incubate at 37°C for 4-6 min (until 90% of the cells have detached). tap flask gently to ensure cells are detached. Add 2% FBS in DPBS at 1 mL / 25 cm² to neutralize the trypsin. Centrifuge cells at 1000rpm for 5 min at room temperature. Remove supernatant. Resuspend pellet in 6.0 to 8.0 mL Complete Growth Medium. Count cells, and seed 4.0 x 10³ to 6.0 x 10³ viable cells/cm² to new culture vessels. Medium Renewal: Every 2-3 days. Note: The cells are sensitive to DMSO and FBS. Ensure that as much as possible is removed after centrifugation, and before seeding into the recommended culture medium.
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
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
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

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