Preparation and characterization of Cryptococcus neoformans synchronous culture

We have developed a method for preparation of synchronous culture in Cryptococcus neoformans. The method is based on age fractionation of exponentially growing asynchronous culture through differential sedimentation in 10–20% (w/v) lactose gradient. C. neoformans capsule thickness should be reduced to a minimum to ensure most accurate age fractionation, which is necessary to obtain a higher degree of synchrony. The C. neoformans synchronous culture system has revealed important characteristics with respect to cellular morphology, DNA content and cell volume distribution. The method can be used for further cell cycle studies. [Copyright &y& Elsevier]