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微信小章| 产品名称： | CuFi-4 |
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| 商品货号： | TS152414 |
| Organism： | Homo sapiens, human |
| Tissue： | Lung; bronchus |
| Cell Type： | Epithelial cells immortalized with hTERT, HPV-16 E6/E7-LXSN, and pBABE-Hyg-TERT |
| 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： | 33 years |
| Gender： | Female |
| Applications： | This cell line may be a useful model for studying ion channel physiology, therapeutic interventions for cystic fibrosis and innate immunity. |
| Storage Conditions： | Liquid nitrogen vapor phase |
| Karyotype： | The karyotypes of several different passages were determined. This is a human cell line of female origin, and the ploidies range from near-diploid to near-tetraploid. The karyology seems to stabilize at higher passages in the hyperdiploid range with trisomies or tetrasomies of chromosomes 1, 5, 8, 11 and 20. Additional copies of chromosomes 5 and 20 were the most consistent aberrations found throughout all the passages and ploidies. |
| Images： |  |
| Derivation： | Human airway epithelial (HAE) cell line, CuFi-4, was derived from lung of a 33-year-old patient with cystic fibrosis by dual retroviral infection with HPV-16E6/E7-LXSN and pBabe-hygro-hTERT.xa0 |
| Antigen Expression： | Antigen expression: This cell line is positive for epithelial marker pan-cytokeratin as assessed by immunocytochemistry. |
| Comments： | CuFi-4 cells are heterozygous for the cystic fibrosis-causing mutations delta F508/G551D. When seeded on semi-permeable filters and cultured at an air-liquid interface, they are capable of forming polarized, differentiated epithelia that exhibit transepithelial resistance and maintain the ion channel physiology expected of a bronchial epithelium.xa0 |
| 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： | xa0 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 CSF1PO: 13 D13S317: 11,12 D16S539: 12,13 D5S818: 11,13 D7S820: 10,12 THO1: 7 TPOX: 8,11 vWA: 17,18 |
| 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 Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332 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 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. |