Hematopoiesis on nylon mesh templates. I. Long-term culture of rat bone marrow cells

Rodent hematopoietic cells have been perpetuated in long-term bone marrow culture (LTBMC), provided that they were plated onto a pre-established layer of bone marrow stromal cells (fibroblasts, reticular cells, adipocytes, macrophages, etc.). This monolayer-type system supports the self-renewal of murine pluripotent stem cells and produces substantial numbers of progenitors and mature cells of the myeloid lineage. In an effort to increase the growth potential of cells in LTBMC, an adherent matrix of stromal cells was established on a pretreated nylon screen template. Subsequent seeding of hematopoietic cells onto this matrix occurred both on the surface of the adherent layer and in the interstices formed by the developing stromal cells. A three dimensional growth pattern of hematopoietic colonies and clusters was observed. Mature cells and late stage precursors of the myeloid and erythroid series were observed in the non-adherent layer for the duration of the experiment (39 weeks). Cells similar in appearance to small lymphocytes were also seen in both the non-adherent and adherent layers. Cells associated with the adherent layer of this LTBMC system displayed the ability to reconstitute hematopoiesis in the irradiated host. It is suggested that because of its three-dimensional nature the nylon mesh LTBMC system possesses a greater hematologic potential per unit area than monolayer-type LTBMC.