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微信小章| 产品名称： | CuFi-6 |
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| 商品货号： | TS171779 |
| Organism： | Homo sapiens, human |
| Tissue： | Lung; bronchus |
| Cell Type： | Epithelial immortalized with hTERT and HPV-16 E6/E7-LXSN |
| Product Format： | frozen |
| Morphology： | Epithelial |
| Culture Properties： | Adherent |
| Biosafety Level： | 2 xa0Cells contain SV40 and HPV viral DNA 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： | Cystic fibrosis |
| Age： | 20 years |
| Gender： | Male |
| Storage Conditions： | Liquid nitrogen vapor phase |
| Karyotype： | This is a near-diploid cell line of male origin. The most consistent karyotypic aberrations include trisomy of chromosome 5 and 20, monosomy of chromosome 15 or 16, and loss of the Y chromosome. Additionally, the polyploidy rate may increase slightly at high passage. |
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| Derivation： | Human airway epithelial (HAE) cell line, CuFi-6, was derived from lung of a 20-year-old patient with cystic fibrosis by retroviral infection with HPV-16 E6/E7-LXSN.xa0 |
| Antigen Expression： | Positive for the epithelial marker pan-cytokeratin as assess by immunocytochemistry. |
| Comments： | CuFi-6 cells are homozygous for the delta F508 cystic fibrosis-causing mutation (delta F508/delta F508).
The cells do not undergo growth arrest in cell culture due to exogenous expression of the telomerase and E6/E7 genes.
Another hTERT-immortalized cell line derived from cystic fibrosis HAE is available as ATCC CRL-4013 (CuFi-1). |
| Complete Growth Medium： | These cells are grown in a serum-free medium: BEGM (Bronchial Epithelial Growth Medium, Serum-free) from Lonza (BEGM Bullet Kit; CC-3170) made of BEBM basal medium and SingleQuot additives (ATCC does not use gentamycin-amphotericin B) supplemented with 50 µg/ml G-418.
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| Subculturing： | Note: The culture flasks should be pre-coated with 60 µg/mL solution of Human Placental Collagen Type IV. (Sigma, Cat. No. C-7521) at least 18 hours in advance then air-dried and rinsed 2-3 times with Dulbeccos Phosphate Buffered Saline. Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Subcultivation ratio: A subcultivation ratio of 1:2 to 1:3 is recommended.
Medium renewal: every 3 to 4 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Aminal Cells: A Manual of Basic Techniques by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005. |
| Cryopreservation： | Freeze medium BEGM supplemented with 10% (v/v) DMSO and 30% (v/v) fetal bovine serum Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions： | Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| STR Profile： | Amelogenin: XY
CSF1PO: 11 D13S317: 12 D16S539: 11,12 D5S818: 9,12 D7S820:8,11 THO1: 6 TPOX: 8,10 vWA: 17,19 |
| 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： | AJ Klingelhutz |
| Year of Origin： | 2001 |
| References： | Halbert CL, et al. The E7 gene of human papillomavirus type 16 is sufficient for immortalization of human epithelial cells. J. Virol. 65: 473-478, 1991. PubMed: 1845902 Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332 Zabner J, et al. Development of cystic fibrosis and noncystic fibrosis airway cell lines . Am. J. Physiol. Lung Cell Mol Physiol. 284: L844-L854, 2003. PubMed: 12676769 Kiyono T, et al. Both Rb/p161NK4a inactivation and telomerase activity are required to immortalize human epithelial cells. Nature 396: 84-88, 1998. PubMed: 9817205 Freshney RI. Culture of Animal Cells: A Manual of Basic Technique, 5th edition. New York: Wiley Liss; 2005. For more information on enzymatic dissociation and subculturing of cell lines see Chapter 13. |