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NuLi-1
NuLi-1
規格:
價格:
編號:B172302
品牌:Mingzhoubio

標準菌株
定量菌液
DNA
RNA

規格:
凍干粉
斜面
甘油
平板


產品名稱
NuLi-1
商品貨號
B172302
Organism
Homo sapiens, human
Tissue
Lung; epithelium, bronchus
培養基
Cell Type
Epithelial cells immortalized with E6/E7 and hTERT
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
Normal
Age
36 years
Gender
Male
Applications
These cell lines may be useful models for studying ion physiology, therapeutic interventions for cystic fibrosis and innate immunity. 
Storage Conditions
Liquid nitrogen vapor phase
Karyotype
This is a near-diploid human cell line of male origin with a polyploidy rate of 24%. There were copies of karyotypically normal X and Y-chromosomes present in most of the cells analyzed. Overall, some of the cells contained chromosomal abnormalities, with the most consistent being trisomy 5 and 20.
Images
Derivation
Human airway epithelial (HAE) cell line, NuLi-1, was derived from normal lung of a 36-year-old patient by dual retroviral infection with HPV-16E6/E7-LXSN [Pubmed: 1845902] and hTERT-LXSN [Pubmed: 9817205]. 
Genes Expressed
NuLi-1 cells (wild type) do not express the delta F508 cystic fibrosis-causing mutation
Comments

The cells do not undergo growth arrest in cell culture due to exogenous expression of the telomerase and HPV-16 E6/E7 genes. 

Another hTERT-immortalized cell line, derived from HAE of a cystic fibrosis patient is available as ATCC® CRL-4013™ (CuFi-1).  Both cell lines, when seeded on semi-permeable 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. 

At this time, ATCC does not include functionality tests like those outlined in Zabner et al [Pubmed: 12676769].

Complete Growth Medium
These cells are grown in a serum-free medium: Airway Epithelial Cell Basal Medium (ATCC PCS-300-030) and Bronchial Epithelial Cell Growth Kit (ATCC PCS-300-040) additives.
Atmosphere: air, 95%; CO2, 5%
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 or equivalent) at least 18 hours in advance then air-dried and rinsed 2-3 times with Dulbecco's Phosphate Buffered Saline. Volumes are given for a 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Subculture when the culture is about 90% confluence. Expected cell yield is between 4 x 104 and 5 x 104 viable cells/cm2.
  2. Remove and discard culture medium.
  3. Add 2.0 to 3.0 mL of 0.25% (w/v) trypsin-0.53 mM EDTA solution to the flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 5 to 10 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Place at 37°C to facilitate dispersal.
  4. To stop trypsinization, add 2.0 to 3.0 mL of 1% FBS in Dulbecco's Phosphate Buffered Saline and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a 15 mL centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
  6. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1.5 to 2.0 x 104 viable cells/cm2 is recommended.
  7. Incubate cultures at 37°C.
Subcultivation Ratio: 1:3
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 Animal Cells: A Manual of Basic Techniques by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
Cryopreservation
Freeze medium: Complete culture medium + 30% FBS + 10% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: 5% CO2 in air recommended
Temperature: 37°C
STR Profile
CSF1PO: 11
D13S317: 10,12
D16S539: 8,13
D5S818: 11,13
D7S820: 12,13
TH01: 9.3
TPOX: 8,11
vWA: 14,18
Amelogenin: X,Y
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
Deposited As
Homo sapiens
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.

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