cAMP included during cell suspension preparation improves survival of dopaminergic neurons in vitro

The physical process of cell suspension preparation from embryonic ventral mesencephala (VM) may be responsible for the low numbers of dopaminergic (DA) neurons that survive following neural transplantation or in vitro culture. In particular, the disruption of cell to extracellular matrix attachment may result in cell death through deactivation of a cAMP-dependent protein kinase involved in cell survival signalling. In an attempt to reduce this death, dibutyryl cAMP was included in all solutions from explant collection to final dissociation. Pretreatment with 700 microM dibutyryl cAMP resulted in 90% survival of the DA neurons originally plated, compared with only 40% in the untreated cultures, after 5 days in vitro. Treatment of VM explants in this manner may result in major improvements in neural transplantation as a technique for the treatment of Parkinson's disease.