Water-based ferrofluids (magnetic fluids) with double-layer steric stabilization by short monocarboxylic acids (lauric and myristic acids) are considered to be a potential source of magnetic nanoparticles in brain cancer (glioblastoma) treatment. Structure characterization in the absence of an external magnetic field is performed, including transmission electron microscopy, magnetization analysis, and small-angle neutron scattering with contrast variation. It is shown that despite the good stability of the systems a significant part of the magnetite nanoparticles are in aggregates, whose inner structure depends on the stabilizer used. In particular, an incomplete coating of magnetite particles is concluded in the case of myristic acid stabilization. The ferrofluids keep their structure unchanged when added to the cancer cell medium. The intracellular accumulations of magnetite from the ferrofluids added to cancer cell cultures as well as its cytotoxicity with respect to human brain cells are investigated.