A new preadipose cell line derived from newborn mouse calvaria can promote the proliferation of pluripotent hemopoietic stem cells in vitro

Abstract

A clonal preadipose cell line MC3T3-G2/PA6, established from newborn mouse calvaria, responds to glucocorticoids and converts to adipose cells in a fashion similar to bone marrow preadipocytes. We investigated the effect of the cells on in vitro hemopoiesis of mouse bone marrow cells by cocultivation. When bone marrow cells were inoculated into confluent cultures of MC3T3-G2/PA6 cells (10(4)-10(6) cells/25-cm2 flask), the number of hemopoietic stem cells (CFU-S) significantly increased during 7-day cultivation in proportion to inoculum size. Under these conditions, active replication of CFU-S was maintained for several weeks until MC3T3-G2/PA6 cell layers detached from the substratum. This capacity of the MC3T3-G2/PA6 line was unique because other established cell lines, including the MTF preadipose line, failed to support CFU-S growth. When bone marrow cells were not allowed to contact the MC3T3-G2/PA6 cell layer, only a small number of CFU-S survived for 7 days. Moreover, MC3T3-G2/PA6 cell-conditioned medium did not show any growth-promoting activity for CFU-S. These results indicate that the MC3T3-G2/PA6 cell line has the ability to promote the proliferation of CFU-S through a short range cell-to-cell interaction by providing an in vitro microenvironment probably similar to that for in vivo hemopoiesis.