STUDY OF THE IMPACT OF THE PRO-INFLAMMATORY RESPONSE ON THE DIFFERENTIATION OF HUMAN DOPAMINERGIC PRECURSORS

MARIA FARIAS; XIANMIN ZENG; VICTORIA GRADASCHI ; CARINA FERRARI; SHIRLEY D WENKER; CORINA GARCIA; FERNANDO PITOSSI

Buenos Aires

Congreso; Reunión Conjunta de Sociedades Biomédicas; 2017

SOCIEDAD ARGENTINA DE INVESTIGACIÓN CLÍNICA

Parkinson ´s Disease (PD) is a neurodegenerative disorder characterizedby the loss of dopaminergic (DA) neurons of the substantianigra pars compacta. Transplantation of dopamine-producing cellsinto the striatum has already shown its effectiveness in animal modelsand clinical trials. Here, we aimed to study the host primary responserelated to the graft of human DA precursors (DA14) in vivoand the impact of pro-inflammatory factors on the viability and differentiationprocess of DA14 cells in vitro. DA differentiation protocolfrom human neural stem cells was standardized and characterizedby immunofluorescence for the following markers: beta III Tubulin(neuronal cells), GFAP (astroglial cells), Foxa2 (DA precursors) andTyrosine hydroxylase (TH) (DA neurons). We analysed cell populationat two stages: DA14 (Foxa2: 71,5%; TH: 5,2%) and final stage,DA28 (beta III Tubulin: 59,4%; TH: 17,4%; GFAP: 3,9%). For invivo assay, DA14 were transplanted into the striatum of adult immunosuppressedrats and host-primary response was analysed byimmunofluorescence. Preliminary results showed host-MHCII andGFAP positive cells within the graft after 7 days of surgery. In orderto study the effect of microglial activation on DA14 maturation invitro, BV2 microglial cells were activated with bacterial lipopolysaccharides(LPS). DA14 cells were then exposed to conditioned media(CM) from basal and activated BV2 cells for 4 days and TH positivecells were determined at DA28 stage. TH cell counting revealed thatexposure of DA14 to CM from activated microglia decreased thenumber of DA neurons at final stage (P