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The WI-38 human diploid cell line was derived by Leonard Hayflick from normal embryonic （3 months gestation） lung tissue. [22194] WI-38 cells have a finite lifetime of 50 plus or minus 10 population doublings with a doubling time of 24 hours. [22197] This line was the first human diploid cell line to be used in human vaccine preparation. [26220] 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.
ATCC 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%. Atmosphere: air, 95%; carbon dioxide （CO2）, 5% Temperature: 37.0°C
Subculturing:
Trypsin and medium should be prewarmed to 37C and the pH should be adjusted to 7.5 before use. Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature （or at 37C） until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks. Subc*tion Ratio: A subc*tion ratio of 1:2 to 1:4 is recommended Medium Renewal: 2 to 3 times per week
Preservation:
growth medium, 95%; DMSO, 5%
Doubling Time:
24 hrs
Related Products:
Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2003 recommended serum:ATCC 30-2020
References:
21405: 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 22194: Hayflick L, Moorhead PS. The serial c*tion of human diploid cell strains. Exp. Cell Res. 25: 585-621, 1961. PubMed: 13905659 22197: Hayflick L. The limited in vitro lifetime of human diploid cell strains. Exp. Cell Res. 37: 614-636, 1965. PubMed: 14315085 26220: Hayflick L, et al. Preparation of poliovirus vaccines in a human fetal diploid cell strain. Am. J. Hyg. 75: 240-258, 1962. PubMed: 13905660 26223: . First International Conference on Vaccines against Viral and Rickettsial Diseases of Man. Pan Am. Health Organ. Off. Doc. 147: 581, 1967. 29375: 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 32244: 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 32288: 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 32582: 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
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