Biopolym. Cell. 1997; 13(1):70-74.
Cell Biology
Gentle and effective mehod to permeabilize the animal and human cell membrane
1Negrutskii B. S.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680


The permeabilized cell system appears one of the most promising methods to study recently discovered subcellular organization of some metabolic processes and the functional mechanisms involved. The existing permeabilization procedures offer rather harsh membrane treatment which leads to the loss of cell integrity. The plant glycoside saponin was used to permeabilize human and animal cells in this work. The saponin treatment is found to provide both essential conservation of intracetlular organization and unlimited access of exogenous macromolecutes to the inside of cell. The permeabilization conditions are slightly different for immobilized cells and cells in suspension. High functional activity of permeabilized cells as well as the availability of the interior of cells to fluorescently labeled macromolecules are demonstrated.


[1] Srere PA. Complexes of sequential metabolic enzymes. Annu Rev Biochem. 1987;56:89-124.
[2] Spivey HO, Merz JM. Metabolic compartmentation. Bioessays. 1989;10(4):127-30.
[3] Negrutskii BS, Deutscher MP. Channeling of aminoacyl-tRNA for protein synthesis in vivo. Proc Natl Acad Sci U S A. 1991;88(11):4991-5.
[4] Heppel LA, Makan N. Methods for rapidly altering the permeability of mammalian cells. J Supramol Struct. 1977;6(3):399-409.
[5] Ryazanov AG, Ovchinnikov LP, Spirin AS. Development of structural organization of protein-synthesizing machinery from prokaryotes to eukaryotes. Biosystems. 1987;20(3):275-88.
[6] Melan MA, Sluder G. Redistribution and differential extraction of soluble proteins in permeabilized cultured cells. Implications for immunofluorescence microscopy. J Cell Sci. 1992;101 ( Pt 4):731-43.
[7] Newmeyer DD, Finlay DR, Forbes DJ. In vitro transport of a fluorescent nuclear protein and exclusion of non-nuclear proteins. J Cell Biol. 1986;103(6 Pt 1):2091-102.
[8] Jacob MC, Favre M, Bensa JC. Membrane cell permeabilization with saponin and multiparametric analysis by flow cytometry. Cytometry. 1991;12(6):550-8.
[9] Bangham AD, Horne RW, Glauert AM, Dingle JT, Lucy JA. Action of saponin on biological cell membranes. Nature. 1962;196:952-5.
[10] Wassler M, Jonasson I, Persson R, Fries E. Differential permeabilization of membranes by saponin treatment of isolated rat hepatocytes. Release of secretory proteins. Biochem J. 1987;247(2):407-15.
[11] Lin A, Krockmalnic G, Penman S. Imaging cytoskeleton--mitochondrial membrane attachments by embedment-free electron microscopy of saponin-extracted cells. Proc Natl Acad Sci U S A. 1990;87(21):8565-9.
[12] Voelker DR. Characterization of phosphatidylserine synthesis and translocation in permeabilized animal cells. J Biol Chem. 1990;265(24):14340-6.
[13] Burgess GM, McKinney JS, Fabiato A, Leslie BA, Putney JW Jr. Calcium pools in saponin-permeabilized guinea pig hepatocytes. J Biol Chem. 1983;258(24):15336-45.