Survival and distribution of transplanted human IMR-32 neuroblastoma cells

Abstract

The visualisation of transplanted cell lines is essential to determine both their viability and possible functional properties. Fluorescent latex microspheres were used to label cultured human neuroblastoma IMR-32 cells prior to transplantation. IMR-32 cells were first rendered amitotic by treatment with mitomycin C and bromodeoxyuridine and subsequently incubated with fluorescent microspheres for 3 days. Cell suspensions were prepared from these cultures and transplanted into the cortex and hippocampus of male Sprague-Dawley rats bearing ibotenate lesions of forebrain cholinergic projections. The animals were perfused at 4, 8 and 12 weeks post-transplantation and tissue was prepared for electron and light microscopy. IMR-32 cells containing fluorescent microspheres were clearly visualised in cryostat sections at all time points. Greater survival was seen in the hippocampus, with evidence of migration of cells from the site of implantation. Macrophages were seen at the electron and light microscope level, and were distinct from the discrete fluorescent labelled IMR-32 cells. Ultrastructurally, transplanted IMR-32 cells resembled cells in vitro, with microspheres clearly distinguished within the cytoplasm. Fluorescent microspheres provide a simple and direct labelling technique suitable for long-term transplant experiments using characterised cell lines.