The Properties of hNT Cells Following Transplantation into the Subventricular Zone of the Neonatal Forebrain

Abstract

Neurons derived from the human teratocarcinoma cell line (hNT) establish structural polarity and a fully mature phenotype following transplantation into the rodent brain. Here we describe the transplantation of hNT cells into the anterior part of neonatal subventricular zone (SVZa), which is a prolific region of neuronal progenitor cells. Ordinarily, the progeny of endogenous or homotopically transplanted SVZa cells migrate to the olfactory bulb (OB) along a restricted pathway, the rostral migratory stream (RMS), and differentiate into interneurons. To compare the phenotype of cultured hNT cells to their transplanted cohorts, hNT cells labeled by the fluorescent dye PKH26 were cultured for 1 day and stained with cell-type-specific antibodies. Clusters as well as individual hNT cells were immunoreactive for TuJ1, an antibody that recognizes neuron-specific class III β-tubulin. The distribution and phenotype of the transplanted hNT cells were examined. The majority of transplanted PKH26-labeled hNT cells were found at their site of implantation in the SVZa, while a small proportion of the transplanted hNT cells was situated in the migratory pathway leading to the OB and in the subependymal zone and granule cell layer of the olfactory bulb. Many of the transplanted hNT cells, both within the SVZa and within the RMS, revealed a neuronal phenotype. Collectively, these results reveal the capacity of hNT cells to respond, at least partially, to cues that ordinarily govern the migration of SVZa-derived cells and maintain their neuronal identity.