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

BACKGROUND: In vitro model studies are becoming increasinglypopular for experimental research designs. They includeisolation and expansion of cells of a particular tissue, suchas the nervous tissue which contributes to understanding theunderlying mechanisms in many pathologies. It enables thescrutinization of intracellular signaling pathways responsiblefor cell death. OBJECTIVES: In the literature, there are differentmethods for the isolation and culture of rat embryonic corticalneurons. However, this study developed a feasible, rapidand easily performable method. METHODS: Isolation of neuronswas performed without using enzymatic digestion. Primarycortical cultures neurite outgrowth and neuron numbers perfield of common mediums were compared for neuronal cellsisolation and expansion. In this study, three different culturemediums 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 obtainedwith the current method. The best neuronal growth wasachieved by Medium I, while Medium II and III had moderateeffect on the neurite outgrowth. CONCLUSIONS: Enzyme-freetreatment was introduced and Medium I was used as an alternativemethod for optimal neuron isolation and expansion. Theneuronal cultures are similar to nervous tissue in physiologicalaspects. Hence, Medium I is more similar to the in vivo conditioncompared to Mediums II and III.

Keywords:

cortical neurons , isolation , primary cell culture , rat