Research Project
3495903
A physiological three-dimensional long-term culture system has been developed for the growth of bone marrow. This system involves the establishment of stromal cells on a suspended nylon filtration screen, the growth of stromal cells to sub-confluence, and the inoculation of stroma with fresh or cryopreserved bone marrow cells. Stromal elements distribute themselves throughout the meshwork, secrete matrix proteins and establish an intricate microenvironment for the support of hematopoiesis. Active production of myeloid and erythroid cells is seen for at least twelve weeks. Preliminary analysis of this system reveals the adherent zone to be actively proliferating and consisting of multilineage immature cells. This adherent layer supports active proliferation of CFU-GM, CFU-Gemm, and erythroid progenitor cells, as indicated by standard methylcellulose and plasma clot assay systems. As in the in vivo state the non-adherent zone contains mostly mature cells or late-stage precursors. Multilineage hematologic expression in adherent and non-adherent zones has been confirmed by cytofluorographic analysis and standard differential counts. These findings indicate that the three- dimensional nylon mesh system is a novel in vitro method which the three- dimensional nylon mesh system is a novel in vitro method which supports the growth of several hematologic lineages concurrently as well as the proliferation of various rat, monkey, and human bone marrow stem cells.
