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NCTC clone 929 [L cell, L-929, derivative of Strain L]

(ATCC® CCL-1™)
Organism  Mus musculus 
Tissue  subcutaneous connective tissue; areolar and adipose 
Product Format  frozen 
Morphology  fibroblast 
Culture Properties  adherent 
Biosafety Level  1
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.
Age  100 days 
Gender  male 
Strain  C3H/An 
Applications  This cell line can be used for toxicity testing.
This cell line is a suitable transfection host.
Storage Conditions  liquid nitrogen vapor phase 

Karyotype  modal chromosome number = 66; range = 65 to 68. There were approximately 20 to 30 marker chromosomes present in each metaphase spread. A high percentage of those markers were common to most analyzed cells. A long metacentric chromosome with secondary constriction was noted in 77/100 cells. Note: Cytogenetic information is based on initial seed stock at ATCC. Cytogenetic instability has been reported in the literature for some cell lines. 
Images   
Derivation  The parent L strain was derived from normal subcutaneous areolar and adipose tissue of a 100-day-old male C3H/An mouse.
NCTC clone 929 (Connective tissue, mouse) Clone of strain L was derived in March, 1948. Strain L was one of the first cell strains to be established in continuous culture, and clone 929 was the first cloned strain developed.

Clone 929 was established (by the capillary technique for single cell isolation) from the 95th subculture generation of the parent strain.
Clinical Data  male
mouse
100 days 
Antigen Expression  
Tumorigenic  Yes, in immunosuppressed mice 
Effects  Yes, in immunosuppressed mice
Virus Susceptibility  Vesicular stomatitis, Glasgow (Indiana)
Vesicular stomatitis, Orsay (Indiana)
Encephalomyocarditis virus
Virus Resistance  poliovirus 1, 2, 3; coxsackievirus B5; polyomavirus
Comments  This cell line has been tested and found negative for ectromelia virus (mousepox).
Complete Growth Medium  The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: horse serum to a final concentration of 10%.
Subculturing  Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
1.Remove and discard culture medium.
2.Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3.Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37℃ to facilitate dispersal.
4.Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5.Add appropriate aliquots of the cell suspension to new culture vessels.
6.Incubate cultures at 37℃.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation  Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions  Atmosphere: Air, 95%; carbon dioxide (CO2), 5%
Temperature: 37℃

Name of Depositor  WR Earle 
Year of Origin  March, 1948 
References  Kazazian HH Jr., et al. Restriction site polymorphism in the phosphoglycerate kinase gene on the X chromosome. Hum. Genet. 66: 217-219, 1984. PubMed: 6325324

Fisher EM, et al. Homologous ribosomal protein genes on the human X and Y chromosomes: escape from X inactivation and possible implications for Turner syndrome. Cell 63: 1205-1218, 1990. PubMed: 2124517

Sanford KK, et al. The growth in vitro of single isolated tissue cells. J. Natl. Cancer Inst. 9: 229-246, 1948.

Sugarman BJ, et al. Recombinant human tumor necrosis factor-alpha: effects on proliferation of normal and transformed cells in vitro. Science 230: 943-945, 1985. PubMed: 3933111

ASTM International Standard Practice for Direct Contact Cell Culture Evaluation of Materials for Medical Devices. West Conshohocken, PA:ASTM International;ASTM Standard Test Method F 0813-07.

ASTM International Standard Test Method for Agar Diffusion Cell Culture Screening for Cytotoxicity. West Conshohocken, PA:ASTM International;ASTM Standard Test Method F 0895-84 (Reapproved 2006).

U.S. Pharmacopeia General Chapters:Biological Reactivity Tests, in vitro. Rockville, MD: U.S. Pharmacopeia; USP USP34-NF29, 2011

Westfall BB, et al. The glycogen content of cell suspensions prepared from massive tissue culture: comparison of cells derived from mouse connective tissue and mouse liver. J. Natl. Cancer Inst. 14: 655-664, 1953. PubMed: 13233820

Earle WR, et al. Production of malignancy in vitro. IV. The mouse fibroblast cultures and changes seen in the living cells. J. Natl. Cancer Inst. 4: 165-212, 1943.

Earle WR, et al. The influence of inoculum size on proliferation in tissue cultures. J. Natl. Cancer Inst. 12: 133-153, 1951. PubMed: 14874126

Sanford KK, et al. The tumor-producing capacity of strain L mouse cells after 10 years in vitro. Cancer Res. 16: 162-166, 1956. PubMed: 13293658

Westfall BB, et al. The arginase and rhodanese activities of certain cell strains after long cultivation in vitro. J. Biophys. Biochem. Cytol. 4: 567-570, 1958. PubMed: 13587550

Papkoff J. Regulation of complexed and free catenin pools by distinct mechanisms. J. Biol. Chem. 272: 4536-4543, 1997. PubMed: 9020180

Hu SX, et al. Development of an adenovirus vector with tetracycline-regulatable human tumor necrosis factor alpha gene expression. Cancer Res. 57: 3339-3343, 1997. PubMed: 9269991

Yasin B, et al. Susceptibility of Chlamydia trachomatis to protegrins and defensins. Infect. Immun. 64: 709-713, 1996. PubMed: 8641770

Biological evaluation of medical devices. Part 5: Tests for in vitro cytotoxicity. Sydney, NSW, Australia:Standards Australia;Standards Australia AS ISO 10993.5-2002.

Plastics collapsible containers for human blood and blood components--Part 1: Conventional containers. Annex C. Biological tests.. Geneva (Switzerland):International Organization for Standardization/ANSI;ISO ISO 3826-1:2003.

Dentistry--Preclinical evaluation of biocompatibility of medical devices used in dentistry--Test methods for dental materials. Geneva (Switzerland):International Organization for Standardization/ANSI;ISO ISO 7405:1997.

Biological evaluation of medical devices--Part 5: Tests for in vitro cytotoxicity. Geneva (Switzerland):International Organization for Standardization/ANSI;ISO ISO 10993-5:1999.

Plastic containers for intravenous injection. Geneva (Switzerland):International Organization for Standardization/ANSI;ISO ISO 15747:2003.