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Establishment and characterization of cell liens from the walker carcinoma 256 able to grow in suspension culture and deficient in thymidien kinase

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Summary

A cell line from the Walker carcinosarcoma 256 of the rat has been established in suspension culture in medium with 5% bovien calf serum for over 350 generations, with an average population doubling time of 17 h, a plating efficiency of 56%, a colony forming efficiency of 32%, and a good capacity to form colonies in soft agar. The cells are morphologically indistinguishable from those in the solid tumor and ascites as checked by transmission and scanning electron microscopy. The karyotype is characterized by a modal number of 65 chromosomes and by the presence of a marker metacentric chromosome. The cells express thymidine kinase, λ-glutamyl transpeptidase, and alkaline phosphatase; are aggultinable by concanavalin A; and can be synchronized by the triple thymidine block. They induce primary tumors, both subcutaneously (solid) and intraperitoneally (ascitic), in the rat; are able to metastasize upon injection by the tail vein; and invade the chorioallantoic membrane of the chick embryo. Cells in suspension can be transferred to monolayers, considerably decreasing their tumorigenicity without affecting the other parameters studied, and can be switched back to suspension culture. DNA-mediated transfection showed that DNA from these cells can transform the NIH-3T3 line. Upon growth of the monolayers in a BrdUr-containing medium, a sub-line was establihed that was cloned into a thymidine kinase-deficient line unable to grow, in HAT medium and with properties otherwise similar to those of the parental wild type cells.

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Authors and Affiliations

  1. Departmento de Biologia Celular, Facultad de Ciencias, Universidad Central de Venezuela, Apartado 21201, 1020, Caracas, Venezuela

    Francisco Arvelo, Auri Yabrudi, Maria Elena Delgado & Néstor González-Cadavid

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  1. Francisco Arvelo
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  2. Auri Yabrudi
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  3. Maria Elena Delgado
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  4. Néstor González-Cadavid
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Additional information

This work was supported by grants from the “Consejo Nacional de Investigaciones Cientificas y Tecnologicas' (CONICIT) and from the ‘Consejo de Desarrollo Cientifico y Humanistico’ of the Central University of Venezuela.

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Arvelo, F., Yabrudi, A., Delgado, M.E. et al. Establishment and characterization of cell liens from the walker carcinoma 256 able to grow in suspension culture and deficient in thymidien kinase. In Vitro 20, 549–565 (1984). https://doi.org/10.1007/BF02639771

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