| 產品名稱 | CuFi-1 |
|---|---|
| 商品貨號 | B211175 |
| Organism | Homo sapiens, human |
| Tissue | Lung; epithelium, bronchus |
| Cell Type | Epithelial cells immortalized E6/E7 and hTERT expression |
| Product Format | frozen |
| Morphology | Epithelial-like |
| Culture Properties | Adherent |
| Biosafety Level | 2 [Cells 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 | 14 years |
| Gender | Female |
| Applications | These cell lines may be useful models for studying ion physiology, therapeutic interventions for cystic fibrosis and innate immunity [Pubmed: 12676769]. |
| Storage Conditions | Liquid nitrogen vapor phase |
| Karyotype | This is a near-diploid human cell line of female origin with a modal chromosome count of 46 and a polyploidy rate of 27%. There were two copies of a karyotypically normal X-chromosome present in most of the cells. Overall, some of the cells contained chromosomal abnormalities, with most consistent being trisomy 20. |
| Images |  |
| Derivation | Human airway epithelial (HAE) cell line, CuFi-1, was derived from lung of a 14-year-old patient with cystic fibrosis by dual retroviral infection with HPV-16E6/E7-LXSN [Pubmed: 1845902] and hTERT-LXSN [Pubmed: 9817205]. |
| Comments | The cells do not undergo growth arrest in cell culture due to exogenous expression of the telomerase and E6/E7 genes. CuFi-1 cells are homozygous for the delta F508 cystic fibrosis-causing mutation (delta F508/delta F508). Another hTERT-immortalized cell line, derived from normal HAE is also available as ATCC CRL-4011 (NuLi-1). Both cell lines, when seeded on semipermeable filters and grown at the air-liquid interface, are capable of forming polarized differentiated epithelia that exhibit transepithelial resistance and maintain the ion channel physiology expected of each genotype. |
| 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 Dulbecco?s 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: 1:5
Medium Renewal: Every 2-3 days (do not exceed 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 | Atmosphere: 5% CO2 in air recommended
Temperature: 37°C |
| STR Profile | Amelogenin: X CSF1PO: 12 D13S317: 11,13 D16S539: 11,14 D5S818: 12 D7S820: 8,11 THO1: 6,9.3 TPOX: 8,11 vWA: 17,20 |
| 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 | 2000 |
| 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. |
| 梅經理 | 17280875617 | 1438578920 |
| 胡經理 | 13345964880 | 2438244627 |
| 周經理 | 17757487661 | 1296385441 |
| 于經理 | 18067160830 | 2088210172 |
| 沈經理 | 19548299266 | 2662369050 |
| 李經理 | 13626845108 | 972239479 |
