Interaction of albumin and phospholipid:cholesterol liposomes in growth of Mycoplasma spp

Mycoplasma spp., sterol and fatty acid auxotrophs, are conventionally grown in complex media containing high concentrations of serum. Serum supplies the required lipids, but its presence complicates studies on the metabolism and antigenicity of mycoplasmas as well as the membrane dynamics of these organisms. In the present work, fetal bovine serum was replaced with dilipidated albumin and liposomes containing high concentrations of cholesterol. The liposomes were produced from phosphatidylcholine which contained other lipid species, including phosphatidylethanolamine, phosphatidylglycerol, and cholesterol. Other liposomes containing cholesterol and one phospholipid yielded significantly less growth of Mycoplasma gallisepticum, indicating that several phospholipids are required to achieve growth levels comparable to those obtained with complex medium. The sources and concentrations of cholesterol, albumin, phosphatidylcholine, and other phospholipids and the interactions among them were important affectors of mycoplasmal growth. Optimal lipid and albumin conditions established for M. gallisepticum were then used to propagate five diverse Mycoplasma spp. to growth levels which equalled or surpassed those obtained with medium containing 17% fetal bovine serum.

[1]  J. Slotte,et al.  Serum albumin enhances the uptake of [3H]cholesterol from phosphatidylcholine vesicles by cultured human fibroblasts. , 1984, The Biochemical journal.

[2]  N. Somerson,et al.  Simple staining procedure permits rapid counting of mycoplasma colonies , 1984, Journal of clinical microbiology.

[3]  T. Sasaki,et al.  Utility of egg yolk medium for cultivation of Mycoplasma pneumoniae , 1983, Journal of clinical microbiology.

[4]  T. Sugiyama Medium components adsorbed to mycoplasmal cells. , 1983, The Yale journal of biology and medicine.

[5]  N. Somerson,et al.  Liposomes replace serum for cultivation of fermenting mycoplasmas , 1983, Applied and environmental microbiology.

[6]  A. Rodwell Mycoplasma gallisepticum requires exogenous phospholipid for growth , 1983 .

[7]  D. Jasper,et al.  Characterization of antigens from mycoplasmas of animal origin. , 1983, American journal of veterinary research.

[8]  C. Dahl,et al.  Coordinate regulation of unsaturated phospholipid, RNA, and protein synthesis in Mycoplasma capricolum by cholesterol. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[9]  W. Bredt,et al.  Influence of cell shape and surface charge on attachment of Mycoplasma pneumoniae to glass surfaces , 1983, Journal of bacteriology.

[10]  S. Rottem,et al.  The effect of phospholipase A2 on intact cells and isolated membranes of Mycoplasma gallisepticum. , 1982, European journal of biochemistry.

[11]  C. Nicolau,et al.  Expression of a beta-lactamase activity in Mycoplasma capricolum transfected with the liposome-encapsulated E.coli pBR 322 plasmid. , 1982, Biochemical and biophysical research communications.

[12]  D. Rose,et al.  Isolation of acholeplasmas and a mycoplasma from vegetables , 1982, Applied and environmental microbiology.

[13]  C. le Grimellec,et al.  Control of membrane lipids in Mycoplasma gallisepticum: effect on lipid order , 1981, Journal of bacteriology.

[14]  G. Snell The effects of different concentrations of glucose and horse serum on the growth of Mycoplasma gallisepticum in broth culture. , 1981, Journal of biological standardization.

[15]  N. Somerson,et al.  Cholesterol as a limiting factor in the growth of Mycoplasma pneumoniae , 1980, Applied and environmental microbiology.

[16]  S. Rottem,et al.  Phospholipid and cholesterol uptake by Mycoplasma cells and membranes. , 1980, Biochimica et biophysica acta.

[17]  C. Thorns,et al.  Studies on the effect of growth medium composition on the antigenicity of Mycoplasma bovis , 1980, Journal of Hygiene.

[18]  S. Razin Cholesterol uptake is dependent on membrane fluidity in mycoplasmas. , 1978, Biochimica et biophysica acta.

[19]  F. Melchers,et al.  Complete replacement of serum by albumin, transferrin, and soybean lipid in cultures of lipopolysaccharide-reactive B lymphocytes , 1978, The Journal of experimental medicine.

[20]  S. Razin,et al.  Cholesterol-phosphatidylcholine dispersions as donors of cholesterol to Mycoplasma membranes. , 1977, Biochimica et biophysica acta.

[21]  J. Lucy,et al.  Membrane cholesterol and cell fusion of hen and guinea-pig erythrocytes. , 1977, The Biochemical journal.

[22]  J. Tully,et al.  Pathogenic mycoplasmas: cultivation and vertebrate pathogenicity of a new spiroplasma. , 1977, Science.

[23]  J. Lucy,et al.  Mechanisms of cell fusion , 1975, Nature.

[24]  H. Mcconnell,et al.  Fusion of phospholipid vesicles with viable Acholeplasma laidlawii. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Bradbury,et al.  Studies on the adsorption of certain medium proteins to Mycoplasma gallisepticum and their influence on agglutination and haemagglutination reactions , 1972, Journal of Hygiene.

[26]  M. Kates Techniques of Lipidology , 1972 .

[27]  A. Rodwell The supply of cholesterol and fatty acids for the growth of mycoplasmas. , 1969, Journal of general microbiology.

[28]  C. Panos,et al.  Fatty acid composition, distribution, and requirements of two nonsterol-requiring mycoplasmas from complex but defatted growth media. , 1969, Biochemistry.

[29]  A. Rodwell THE NUTRITION AND METABOLISM OF MYCOPLASMA: PROGRESS AND PROBLEMS , 1967, Annals of the New York Academy of Sciences.

[30]  L. Hayflick Tissue cultures and mycoplasmas. , 1965, Texas reports on biology and medicine.

[31]  W. J. Dyer,et al.  A rapid method of total lipid extraction and purification. , 1959, Canadian journal of biochemistry and physiology.