HEMATOPOIETIC SUPPORTIVE FUNCTION OF HUMAN BONE-MARROW STROMAL CELL-LINES ESTABLISHED BY A RECOMBINANT SV40-ADENOVIRUS VECTOR

We have previously reported the establishment of a variety of human bo ne marrow stromal cell lines using a recombinant SV40-adenovirus vecto r. Using this vector, we obtained more clonal stromal cells. Here, we have characterized these cells and analyzed their capacity to support the proliferation and differentiation of hematopoietic cells. The stro mal cells were cocultured with nonadherent human bone marrow cells use d as hematopoietic cells. The total numbers of hematopoietic cells and CFU-GM in culture were counted every week. Two of the six stromal cel l lines, AA101 and HAS303, supported the proliferation and differentia tion of hematopoietic cells and CFU-GM for more than 9 weeks. Further, granulocytes, macrophages, and megakaryocytes were detected when cocu ltured with these cells. When hematopoietic cells were cocultured but separated from the two stromal cell lines by a 0.45-mu m millipore mem brane to prevent their attachment, almost all CFU-GM disappeared withi n 7 weeks. The supportive Stromal cells produced GM-CSF and IL-6. Howe ver, other cell lines producing these humoral factors did not support hematopoietic cell proliferation for such a long time. These findings suggest that these established human bone marrow stromal cell lines wi ll be useful, in that analysis of their supportive function in hematop oietic cell proliferation and differentiation through cell-to-cell int eraction will shed some light on this area.