Abstract
In this study we show that the ganglioside content and pattern of human skin fibroblasts change along the process of cell subculture progression by varying the cell density.
GM3, GD3 and GD1a were components of the total cell ganglioside mixtures extracted from cells, but GD1a was in all the extracts a minor component or very scant. Other gangliosides present in traces were not characterised. The fibroblast ganglioside content of 52 pools of cells obtained from 5 different cell lines cultured at variable cell density ranged from 2.0 to 13.1 nmoles per mg of cell protein. The molar ratio between GM3 and GD3 varied from 418 to 0.6 in the ganglioside mixtures, as determined by densitometric quantitative analysis after thin layer chromatographic separation.
Both the ganglioside content and the GM3/GD3 molar ratio were constant along several passages of subculture progression performed by plating cells collected at confluence. Instead, when the subculture progression was performed by plating cells collected at a few days after reaching confluence, a progressive increase of the ganglioside content was observed. GD3 increased proportionally more than GM3 so that a progressive decrease of the ratio between GM3 and GD3 was observed. In some experiments, GD3 was very scant at the beginning of the progression, while it was near 30% after 5 passages under these conditions. The progressive increase of GD3 along the high density cell population subculture progression was associated to a moderate increase of the mRNA GD3 synthase. Published in 2003.
Similar content being viewed by others
References
Wiegandt H, New Compr Biochem 10, 199–260 (1985).
Schroeder L, London E, Brown D, Proc Natl Acad Sci USA 91, 12130–4 (1994).
Hakomori S-I, Handa K, Iwabuchi K, Yamamura S, Prinetti A, Glycobiology 8, xi–xix, (1998).
Prinetti A, Iwabuchi K, Hakomori S-I, J Biol Chem 274, 20916–24 (1999).
Kasahara K, Watanabe Y, Yamamoto T, Sanai Y, J Biol Chem 272, 29947–53 (1997).
Dawson G, Matalon R, Dorfman A, J Biol Chem 247, 5944–50 (1972).
Chigorno V, Tettamanti G, Sonnino S, J Biol Chem 271, 21738–44 (1996).
Yogeeswaran G, Hakomiri S-I, Biochemistry 14, 2151–6 (1975).
Ohsawa T, Exp Gerontol 24, 1–9 (1989).
Colombo I, Sottocornola E, Moretti S, Meloni MA, Pippia P, Berra B, Biochim Biophys Acta 1485, 214–24 (2000).
Dohi T, Nores G, Hakomori S-I, Cancer Res 48, 5680–5 (1988).
Acquotti D, Cantu' L, Ragg E, Sonnino S, Eur J Biochem 225, 271–88 (1994).
Hauttecoeur B, Sonnino S, Ghidoni R, Biochim Biophys Acta 833, 303–7 (1985).
Mauri L, Casellato R, Kirschner G, Sonnino S, Glycoconjugate J 16, 197–203 (1999).
Leroy JG, Ho WM, McBrinn MC, Zielke K, Jacob J, O'Brien JS, Pediatr Res 6, 752–7 (1972).
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ, J Biol Chem 193, 265–75 (1951).
Rago R, Mitchen S, Wilding G, Anal Biochem 191, 31–4 (1990).
Tettamanti G, Bonali F, Marchesini S, Zambotti V, Biochim Biophys Acta 296, 160170 (1973).
Partridge SM, Biochem J 42, 238–48 (1948).
Stahl E, Anisaldehyd-Schwefelsaure fur Steroide, Terpene Zucker und so w. Dunnschicht Chromatographie (Springer Verlag, Berlin, 1962), p. 498.
Nobile-Orazio E, Legname G, Daverio R, Carpo M, Giuliani A, Sonnino S, Scarlato G, Ann Neur 28, 190–4 (1990).
Wu G, Ledeen R, Anal Biochem 173, 368–75 (1988).
Taki T, Handa S, Ishiwaka D, Anal Biochem 223, 232–8 (1994).
Svennerholm L, Biochim Biophys Acta 24, 604–11 (1957).
Maggi A, Susanna L, Bettini E, Mantero G, Zucchi I, Mol Endocrinol 3, 1165–70 (1989).
Sambrook J, Russel DW, Molecular Cloning: A Laboratory Manual, 3rd ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2001).
Zucchi I, Montagna C, Susani L, Vezzoni P, Dulbecco R, Proc Natl Acad Sci USA 95, 1079–84 (1998).
Gibbs RA, Nguyen PN, McBride LJ, Koepf SM, Caskey CT, Proc Natl Acad Sci USA 86, 1919–23 (1989).
Svennerholm L, Adv Exp Biol Med 125, 11 (1980)
IUPAC-IUBMB Joint Commission on Biochemical Nomenclature, Pure Appl Chem 69, 2475–87 (1997); Carbohydr Res 312, 167‐75 (1998).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sciannamblo, M., Chigorno, V., Passi, A. et al. Changes of the ganglioside pattern and content in human fibroblasts by high density cell population subculture progression. Glycoconj J 19, 181–186 (2002). https://doi.org/10.1023/A:1024249707516
Issue Date:
DOI: https://doi.org/10.1023/A:1024249707516