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Purification and characterisation of the in vitro colony forming cell in monkey hemopoietic tissue
Author(s) -
Moore M. A. S.,
Williams N.,
Metcalf D.
Publication year - 1972
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1040790213
Subject(s) - in vitro , haematopoiesis , biology , bone marrow , spleen , population , microbiology and biotechnology , lymphocyte , density gradient , thymidine , cell , immunology , stem cell , biochemistry , medicine , physics , environmental health , quantum mechanics
Buoyant density gradient separation of Rhesus monkey bone marrow, spleen and blood leukocytes has demonstrated a reproducible and homogeneous light density distribution profile of cells capable of forming hemopoietic colonies in agar culture ( in vitro colony forming cells — CFC). High resolution density gradient separation performed on a light density fraction of bone marrow produced on average a 100‐fold enrichment of in vitro CFC with the most enriched fractions containing the majority of the in vitro CFC population present in the original marrow. Fractions were routinely obtained in which up to 23% of cells formed colonies and 33% were capable of proliferating to some degree upon stimulation. Tritiated thymidine suiciding showed the active proliferative status of the in vitro CFC and application of autoradiography and morphological characterisation to highly enriched density fractions has shown that the in vitro CFC in normal marrow is a transitional lymphocyte. Single cell transfer experiments have shown that in vitro CFC's formed colonies containing both granulocytes and macrophages, formally demonstrating the clonal origin of in vitro colonies and the common origin of granulocytes and macrophages.