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微信小章| 产品名称： | CuFi-5 |
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| 商品货号： | TS173799 |
| Organism： | Homo sapiens, human |
| Tissue： | Lung; bronchus |
| Cell Type： | Epithelial cells immortalized with hTERT and HPV-16 E6/E7-LXSN |
| Product Format： | frozen |
| Morphology： | Epithelial-like |
| 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： | 32 years |
| Gender： | Male |
| Storage Conditions： | Liquid nitrogen vapor phase |
| Karyotype： | This is a near-diploid cell line of male origin in which the most consistent karyotypic aberrations are trisomy of chromosomes 5 and 20. Other non-clonal aberrations were found at early passage, but the karyology tended to stabilize within several passages. |
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| Derivation： | Human airway epithelial (HAE) cell line, CuFi-5, was derived from lung of a 32-year-old patient with cystic fibrosis by retroviral infection with hTERT and HPV-16E6/E7.xa0 |
| Antigen Expression： | Antigen expression: This cell line is positive for the epithelial marker pan-cytokeratin as assessed by immunocytochemistry. |
| Comments： | CuFi-5 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. Other hTERT-immortalized cell lines derived from cystic fibrosis HAE are also available from ATCC as ATCC CRL-4013 (CuFi-1), ATCC CRL-4015 (CuFi-4), and ATCC CRL-4017 (CuFi-6). |
| 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 60µg/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 2 to 3 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 with 30% FBS and 10% DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions： | Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| STR Profile： | Amelogenin: X,Y CSF1PO: 10,14 D13S317: 11,13 D16S539: 11,13 D5S818: 11,12 D7S820:11,12 THO1: 7 TPOX: 8,10 vWA: 16,17 |
| 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. |
| Name of Depositor： | AJ Klingelhutz |
| Year of Origin： | 2001 |
| References： | 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 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 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. |