One of the most prevalent degenerative disorders of the nervous system is Parkinson's disease. The etiology of this disease is for the most part unknown, although it is posited to arise from an interaction of genetic and environmental factors. Although in vivo animal studies have been used to examine the effects of a number of Parkinson-inducing compounds, there is little information on reliable in vitro methodologies that can recapitulate the previously observed in vivo results. Here, we describe a method for generating mixed and chimeric neuron/glial cultures of postnatal substantia nigra (SN), independent of other monoaminergic nuclei in the ventral midbrain. Since many toxins do not affect regions of the midbrain except the SN, use of whole ventral midbrain from embryos can dilute any measurement of cell death. By specifically culturing ventrolateral midbrain containing the substantia nigra, one can more directly target the effects of dopaminergic toxins. In addition, this method can be used to test potential therapies for amelioration of Parkinson's disease.