T-lymphocyte colony formation by thymic and peripheral lymphocytes in methylcellulose culture.
A new method for one-step and one-layer culture of T-lymphocyte colonies (TLCs) from murine thymocytes and peripheral T-cells in methylcellulose is described. TLCs are detected from day 4 of culture and may contain up to 10(4) cells per colony. As few as 1 X 10(3) cells per culture can be seeded, and presented data indicate that individual colonies are the progenies of single cells. The frequency of colony-forming cells (CFCs) is about one per 6 000 thymic or splenic cells and one per 3 000 lymph node cells. A series of experiments was performed to study the nature of the CFC and the colony-cells, and it is concluded that the CFC is a Thy-1.2. positive, non-adherent, relatively hydrocortisone resistant, radiosensitive T-cell, the vast majority being in G1 or G0 phase, while colony-cells are Thy-1.2. positive, predominantly Lyt-2 positive cells in rapid cell-cycle. This method for growing thymic cells makes it possible to study differentiation and functional restriction of subsets of thymocytes on a clonal basis, and the culture system may also be a valuable source of monoclonally expanded lymphoid cells for future immunological studies.
Röpke
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Claësson
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D000818:Animals; D000950:Antigens, Ly; D000954:Antigens, Surface; D002454:Cell Differentiation; D002455:Cell Division; D002999:Clone Cells / Q000166:cytology; D003584:Cytological Techniques; D007700:Kinetics; D008221:Lymphoid Tissue / Q000166:cytology; D008747:Methylcellulose; D051379:Mice; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology; D018800:Thy-1 Antigens; D013950:Thymus Gland / Q000166:cytology
D000950:Antigens, Ly; D000954:Antigens, Surface; D018800:Thy-1 Antigens; D008747:Methylcellulose