Semaphorins comprise a family of molecules implicated in the guiding of growing axons and neuronal progenitor cells. Further, semaphorins have been suggested to play a role in cancer metastasis. Neuropilins 1 and 2 are cell surface receptors for soluble class 3 semaphorins. Plexins are direct receptors for membrane-bound semaphorins and, by binding to neuropilins, coreceptors necessary for class 3 semaphorin signaling. We here report that human malignant glioma cell lines express neuropilins 1 and 2 mRNA and protein, as well as either plexin A1, A2, or B1. Further, all glioma cell lines express SEMA3A and SEMA3C and exhibit SEMA3A binding sites. Exogenous SEMA3A expressed in 293 or U87MG cells has no collapsing or chemorepulsive activities on glioma cells as determined by F-actin staining and collagen coculture assays. In summary, human glioma cells express class 3 semaphorins and receptors for soluble and membrane-bound semaphorins, suggesting a possible role of the semaphorin/neuropilin system in the interactions of human malignant glioma with the host's central nervous and immune systems. GLIA 42:379–389, 2003. © 2003 Wiley-Liss, Inc.