WI-38 (ATCC® CCL-75)

Organism: Homo sapiens, human  /  Cell Type: fibroblast  /  Tissue: lung  /  Disease: normal

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Organism Homo sapiens, human
Tissue lung
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease normal
Age 3 months gestation fetus
Gender female
Ethnicity Caucasian
Applications
This cell line is a suitable transfection host.
This cell line can be used for viruscide testing.
Storage Conditions liquid nitrogen vapor phase
Karyotype normal diploid
Derivation
The WI-38 human diploid cell line was derived by Leonard Hayflick from normal embryonic (3 months gestation) lung tissue.
Clinical Data
3 months gestation fetus
Caucasian
female
Virus Susceptibility Vesicular stomatitis, Glasgow (Indiana)
Herpes simplex virus
Pseudorabies virus
Human poliovirus 1
Comments
WI-38 cells have a finite lifetime of 50 plus or minus 10 population doublings with a doubling time of 24 hours.

This line was the first human diploid cell line to be used in human vaccine preparation.

The 8th passage ampule from which this freeze was derived was found to contain a bacterial contaminant (a micrococcus). The cell line was subsequently cured by several passages in the presence of antibiotics.

Growth of the cells is enhanced by addition of tumor necrosis factor alpha (TNF alpha) to the medium.

This cell line is negative for reverse transcriptase.

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: fetal bovine serum to a final concentration of 10%.
Subculturing

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium 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-0.53mM EDTA solution to remove all traces of serum which 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°C 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 cell suspension to new culture vessels
  6. Place culture vessels in incubators at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
growth medium, 95%; DMSO, 5%
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
Amelogenin: X
CSF1PO: 10,12
D13S317: 11
D16S539: 11,12
D5S818: 10
D7S820: 9,11
THO1: 8,9.3
TPOX: 8
vWA: 19,20
Isoenzymes
G6PD, B
Population Doubling Time 24 hrs
Name of Depositor L Hayflick
Passage History
The 8th passage ampule from which this freeze was derived was found to contain a bacterial contaminant (a micrococcus). The cell line was subsequently cured by several passages in the presence of antibiotics.
References

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

Hayflick L, Moorhead PS. The serial cultivation of human diploid cell strains. Exp. Cell Res. 25: 585-621, 1961. PubMed: 13905659

Hayflick L. The limited in vitro lifetime of human diploid cell strains. Exp. Cell Res. 37: 614-636, 1965. PubMed: 14315085

Hayflick L, et al. Preparation of poliovirus vaccines in a human fetal diploid cell strain. Am. J. Hyg. 75: 240-258, 1962. PubMed: 13905660

First International Conference on Vaccines against Viral and Rickettsial Diseases of Man. Pan Am. Health Organ. Off. Doc. 147: 581, 1967.

Tomasek JJ, et al. Gelatinase A activation Is regulated by the organization of the polymerized actin cytoskeleton. J. Biol. Chem. 272: 7482-7487, 1997. PubMed: 9054450

Hoppe HC, et al. Identification of phosphatidylinositol mannoside as a mycobacterial adhesin mediating both direct and opsonic binding to nonphagocytic mammalian cells. Infect. Immun. 65: 3896-3905, 1997. PubMed: 9284169

Landers JE, et al. Translational enhancement of mdm2 oncogene expression in human tumor cells containing a stabilized wild-type p53 protein. Cancer Res. 57: 3562-3568, 1997. PubMed: 9270029

Chang K, Pastan I. Molecular cloning of mesothelin, a differentiation antigen present on mesothelium, mesotheliomas, and ovarian cancers. Proc. Natl. Acad. Sci. USA 93: 136-140, 1996. PubMed: 8552591

Debant A, et al. The multidomain protein trio binds the LAR transmembrane tyrosine phosphatase, contains a protein kinase domain, and has separate rac-specific and rho-specific guanine nucleotide exchange factor domains. Proc. Natl. Acad. Sci. USA 93: 5466-5471, 1996. PubMed: 8643598

Zhu X, et al. Cell cycle-dependent modulation of telomerase activity in tumor cells. Proc. Natl. Acad. Sci. USA 93: 6091-6095, 1996. PubMed: 8650224

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

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

Cross References

Nucleotide (GenBank) : AA762049 EST01 Human Lung cDNA Library Homo sapiens cDNA clone 3I35 5', mRNA sequence.

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

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

Hayflick L, Moorhead PS. The serial cultivation of human diploid cell strains. Exp. Cell Res. 25: 585-621, 1961. PubMed: 13905659

Hayflick L. The limited in vitro lifetime of human diploid cell strains. Exp. Cell Res. 37: 614-636, 1965. PubMed: 14315085

Hayflick L, et al. Preparation of poliovirus vaccines in a human fetal diploid cell strain. Am. J. Hyg. 75: 240-258, 1962. PubMed: 13905660

First International Conference on Vaccines against Viral and Rickettsial Diseases of Man. Pan Am. Health Organ. Off. Doc. 147: 581, 1967.

Tomasek JJ, et al. Gelatinase A activation Is regulated by the organization of the polymerized actin cytoskeleton. J. Biol. Chem. 272: 7482-7487, 1997. PubMed: 9054450

Hoppe HC, et al. Identification of phosphatidylinositol mannoside as a mycobacterial adhesin mediating both direct and opsonic binding to nonphagocytic mammalian cells. Infect. Immun. 65: 3896-3905, 1997. PubMed: 9284169

Landers JE, et al. Translational enhancement of mdm2 oncogene expression in human tumor cells containing a stabilized wild-type p53 protein. Cancer Res. 57: 3562-3568, 1997. PubMed: 9270029

Chang K, Pastan I. Molecular cloning of mesothelin, a differentiation antigen present on mesothelium, mesotheliomas, and ovarian cancers. Proc. Natl. Acad. Sci. USA 93: 136-140, 1996. PubMed: 8552591

Debant A, et al. The multidomain protein trio binds the LAR transmembrane tyrosine phosphatase, contains a protein kinase domain, and has separate rac-specific and rho-specific guanine nucleotide exchange factor domains. Proc. Natl. Acad. Sci. USA 93: 5466-5471, 1996. PubMed: 8643598

Zhu X, et al. Cell cycle-dependent modulation of telomerase activity in tumor cells. Proc. Natl. Acad. Sci. USA 93: 6091-6095, 1996. PubMed: 8650224

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

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