Cryopreservation of granule cells from the postnatal rat hippocampus

Although primary cultures of neurons are essential methods for cell biological and pharmacological researches, many animals must be sacrificed for each experiment. Here we introduce a novel system to cryopreserve hippocampal granule cells (GCs) prepared from postnatal rats. Being thawed after as long as 60 days of cryopreservation, GCs expressed the mature neuronal marker MAP-2 and elongated single tau-1-positive axons and multiple tau-1-negative dendrites. These properties closely resembled intact GCs in primary cultures, providing the advantage of being able to repeatedly prepare stable cultures with a single sacrifice of animals.