Density distribution analysis of In vivo and In vitro colony forming cells in bone marrow

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Abstract
The technique of buoyant density separation in gradients of Bovine Serum Albumin has been used to separate hemopoietic cell populations in mouse bone marrow that form in vivo spleen colonies and in vitro colonies of granulocytes and macrophages in an agar culture system. The density distribution profiles showed a number of reproducible density subpopulations of both in vivo and in vitro colony forming cells (C.F.C.'s). The mean density of in vitro C.F.C.'s exceeded that of the in vivo but overlap of the density profiles of the two populations was evident. Density‐related differences in seeding efficiency of in vivo C.F.C.'s were observed.Freund's adjuvant treatment increased marrow and spleen in vitro C.F.C. populations. Marrow density profiles obtained three and seven days after adjuvant showed a progressive increase in in vitro C.F.C.'s in a restricted density region with no associated elevation of in vivo activity.The antimitotic agent, vinblastine, revealed differences in mitotic activity between the two cell populations, reducing the in vitro C.F.C. population to .07% and the in vivo to 5% of normal in 24 hours. Density separation of vinblastine‐treated marrow produced density regions devoid of in vitro activity but containing in vivo in vivo C.F.C.'s which, upon transfer to irradiated recipients, regenerated both in vivo and in vitro density distribution profiles.

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