Microcultures of Dissociated Primary Central Nervous System Neurons

Abstract

This chapter discusses the microcultures of dissociated primary central nervous system (CNS) neurons. The development of a microculture system has been aimed at establishing a large number of separate cultures of primary CNS neurons. This chapter discusses preparation of a solution of dissociated primary cells from brain regions rich in neurons of known neurotransmitter content and to plate these cells onto 96-well plates treated with an artificial positively charged substratum. The neurons are grown under standard tissue culture techniques and biochemical procedures adapted for rapid and sensitive measurements of chemically identified neurons. The microculture approach utilizes multitipped pipetters, small 96-well plate incubators, multiwell plate centrifuges, a Titertek multiskan spectrophotometer, automatic injector systems for high-performance liquid chromatography (HPLC), computer-assisted scintillation counting, and statistical and graphic software analysis. The ability to grow and analyze the maturation of chemically identified primary neurons in microcultures permits routine study of up to 1000 cultures a week by a single research team.