In vitro proliferation of haemopoietic cells in the presence of adherent cell layers. I. Culture conditions and strain dependence.

The culture system, in which a marrow-derived adherent cell population, established in vitro, exerts a long-term promoting influence on proliferation of haemopoietic cells, is reproduced. Essential parameters of the system are investigated; it is confirmed that the system is critically dependent on horse serum, and on the in vitro age of the adherent cell layer. The growth-promoting effect on haemopoietic cells seems to be independent of the number of marrow cells per culture flask initially inoculated into the cultures to establish the adherent cell layer. In vitro established marrow-derived adherent cell layers from RFM (H-2f) and BALB/c (H-2d) mice can promote the long-term proliferation of syngeneic and allogenic haemopoietic cells; haemopoietic marrow cells from C3H (H-2k) cannot be maintained on syngeneic or allogeneic (BALB/c, H-2d) adherent cell layers; adherent cell layers of C3H (H-2k) can maintain haemopoietic cells of the H-2d (BALB/c) genotype. This culture system does not reproduce the in vivo phenomenon of allogeneic resistance. The relevance of these findings to the suggestion that the growth-promoting activity of adherent marrow cells on haemopoietic stem cells in vitro duplicates aspects of the in vivo haemopoietic microenvironment is discussed.