PMID: 6397572Jan 1, 1984Paper

Segregation and characterization of lymphohematopoietic stromal elements

Kroc Foundation Series
D A CrouseJ G Sharp

Abstract

In the mouse, long-term maintenance of multipotent hematopoietic stem cells in vitro currently requires the establishment of an appropriate adherent layer. When established employing culture medium supplemented with 20% horse serum and 10(-6) M hydrocortisone, the initial adherent layer contains primitive stem cells that are the major contributor of multipotent stem cells (CFUs) assayed subsequently in the supernatant regardless of the addition of fresh bone marrow cells. In contrast, when the adherent layer is established employing 25% fetal calf serum without hydrocortisone, few if any stem cells survive in the adherent layer. Such cultures are dependent upon a recharge with fresh bone marrow as a source of CFUs. A comparison of the latter stem cell-depleted system with the former intact system permits an evaluation of the relative contributions of the adherent layer and stem cells to the long-term maintenance of hematopoiesis in vitro. Studies of the effects of irradiation of the donor animal for the adherent layer and using the intact system demonstrated a reduction in the supernatant CFUs production that was dose- and time-related and evident at doses of 100 and 500 rads. If the adherent layer itself was irradiated immedia...Continue Reading

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