Select growth of human T lymphocytes in single phase semisolid culture

J. Shen, F. Wilson, M. Shifrine, M. Eric Gershwin

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Selective growth and clonal proliferation of human T lymphocytes can be achieved by using a single-phase semi-solid methylcellulose system without the requirement of preincubation with lectins. Significant proliferation, however, depends upon the continued presence of Con A or PHA, but not pokeweed mitogen or lipopolysaccharide within the methylcellulose. This procedure eliminates nonspecific agglutination by lectins and allows for direct visualization of colonies and their specific removal and subsequent cloning in liquid phase. Optimal growth and production of colonies greater than 40-cell size require 3 to 9 days. Individual cells can be identified on the basis of E rosette formation and absence of surface immunoglobulin or ability to phagocytize latex particles. Moreover, proliferation is inhibited by antithymocyte but not anti-B cell sera and can be demonstrated with peripheral blood T and MOLT-4 cells, but not with B or Raji cells. Finall, colony formation is not enhanced by the presence of 2-mercaptoethanol. The clonal proliferation of human T lymphocytes has wide application in the study of both antigen recognition and lymphocyte alterations in specific diseases.

Original languageEnglish (US)
Pages (from-to)1299-1305
Number of pages7
JournalJournal of Immunology
Volume119
Issue number4
StatePublished - 1977

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Methylcellulose
T-Lymphocytes
Lectins
Growth
Rosette Formation
Pokeweed Mitogens
B-Cell Antigen Receptors
Mercaptoethanol
Agglutination
Microspheres
Cell Size
Lipopolysaccharides
Organism Cloning
B-Lymphocytes
Lymphocytes
Antigens
Serum

ASJC Scopus subject areas

  • Immunology

Cite this

Select growth of human T lymphocytes in single phase semisolid culture. / Shen, J.; Wilson, F.; Shifrine, M.; Gershwin, M. Eric.

In: Journal of Immunology, Vol. 119, No. 4, 1977, p. 1299-1305.

Research output: Contribution to journalArticle

Shen, J, Wilson, F, Shifrine, M & Gershwin, ME 1977, 'Select growth of human T lymphocytes in single phase semisolid culture', Journal of Immunology, vol. 119, no. 4, pp. 1299-1305.
Shen, J. ; Wilson, F. ; Shifrine, M. ; Gershwin, M. Eric. / Select growth of human T lymphocytes in single phase semisolid culture. In: Journal of Immunology. 1977 ; Vol. 119, No. 4. pp. 1299-1305.
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