Abstract Based on the successful use of a basement membrane-like extracellular matrix (ECM) as a physiological substrate for the growth of adult rat oligodendrocytes, we have applied the system to oligodendroglia of human origin. ECM is excreted and deposited on a culture dish by endothelial cells. Glial cells isolated from fresh human brains by a Percoll density gradient were maintained on ECM for several weeks in vitro. Cells identified as oligodendrocytes by anti-galactocerebroside (GalC) could be transferred and replated on ECM-coated plates. In one case, GalC-positive cells isolated from a biopsy of an 8-year old girl were able to incorporate [ 3H]thymidine as revealed by autoradiography. ECM can be used successfully for the growth and in vitro maintenance of human oligodendrocytes.