Effects of different culture media on optimization of primary neuronal cell culture for in vitro models assay

Abstract

Background: In vitro model studies are becoming increasingly popular for experimental research designs. They include isolation and expansion of cells of a particular tissue, such as the nervous tissue which contributes to understanding the underlying mechanisms in many pathologies. It enables the scrutinization of intracellular signaling pathways responsible for cell death. OBJECTIVES: In the literature, there are different methods for the isolation and culture of rat embryonic cortical neurons. However, this study developed a feasible, rapid and easily performable method. METHODS: Isolation of neurons was performed without using enzymatic digestion. Primary cortical cultures neurite outgrowth and neuron numbers per field of common mediums were compared for neuronal cells isolation and expansion. In this study, three different culture mediums were considered: Medium I: Neurobasal medium, B-27 and L-glutamine; Medium II: DMEM, FBS and L-glutamine; and Medium III: DMEM/F-12, FBS and L-glutamine. RESULTS: High survival rate and number of neurons was obtained with the current method. The best neuronal growth was achieved by Medium I, while Medium II and III had moderate effect on the neurite outgrowth. CONCLUSIONS: Enzyme-free treatment was introduced and Medium I was used as an alternative method for optimal neuron isolation and expansion. The neuronal cultures are similar to nervous tissue in physiological aspects. Hence, Medium I is more similar to the in vivo condition compared to Mediums II and III.