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Cells immortalized by CDK4
Cells immortalized by hTERT
Cells contain SV40 promoter sequences
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
The immortalized HSAEC1-KT cells show a stable epithelial morphology and differentiated cytokeratin isoforms after over 100 population doublings, express the stem cell marker p63 and high levels of p16INK4a, and have an intact p53 checkpoint pathway (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).
The HSAEC1-KT cells also exhibit characteristics of normal cells such as contact inhibition of growth and failure to form soft agar colonies or form tumors in immune compromised mice (RefKalita M, et al. Systems approaches to modeling chronic mucosal inflammation. Biomed. Res. Int. 2013: 505864, 2013. PubMed: 24228254).
TGFβ-treated HSAEC1-KT cells showed an elongated shape with markedly induced F-actin staining. This morphological change of enhanced front-rear polarity and cytoskeletal actin rearrangement are characteristic morphological changes of EMT (epithelial mesenchymal transition) (RefKalita M, et al. Systems approaches to modeling chronic mucosal inflammation. Biomed. Res. Int. 2013: 505864, 2013. PubMed: 24228254).
The HSAEC1-KT cell line was established by infecting primary human small airway epithelial cell culture with human telomerase (hTERT) and mouse cyclin dependent kinase 4 (CDK4) expressing retrovirus constructs and selecting under 250 ng/mL puromycin and 30 ug/mL 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)
Cytogenetic analysis was performed on G-banded metaphase cells from the human cell line HSAEC1-KT. Several abnormal male near-diploid karyotypes are found:
Clone 1 demonstrates trisomy 5 with no other aberrations. 47,XY,+5
Clone 2 demonstrates trisomy 5 and 20 with no other aberrations. 48,XY,+5,+20
Clone 3 demonstrates trisomy 5, an isochromosome of the long-arm of chromosome 10, resulting in three copies of the chromosome 10 long-arm and only one copy of the short-arm, and trisomy 20. 48,XY,+5,i(10)(p10),+20
Clone 4 demonstrates a deletion on the short-arm of chromosome 10 at band p10, a deletion on the short-arm of chromosome 17 at band p13, and trisomy 20. 47,XY,del(10)(p10),del(17)(p13),+20.
To make the complete culture medium, add SAGM™ SingleQuots™ (Lonza CC-4124) which contains supplements and growth factors (BPE, Hydrocortisone, hEGF, Epinephrine, Transferrin, Insulin, Retinoic Acid, Triiodothyronine, BSA-FAF) to 500 mL bottle of SABM Basal Medium™, phenol red free basal medium (Lonza CC-3119)
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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
Kalita M, et al. Systems approaches to modeling chronic mucosal inflammation. Biomed. Res. Int. 2013: 505864, 2013. PubMed: 24228254
Gazdar AF, Gao B, Minna JD. Lung cancer cell lines: Useless artifacts or invaluable tools for medical science? Lung Cancer 68(3): 309-318, 2010. PubMed: 20079948