CRYOPRESERVATION OF RAT NEURONAL CELLS BY SLOW COOLING METHODOLOGY

JUAN DE PAZ L.; M. CELESTE ROBERT; BERARDI F.; BALABAN C.L.; GRAF D.; SILVIA GAZZIN; CLAUDIO TIRIBELLI; RODRIGUEZ J.V.

Viña del MAr

Taller; 3ER Taller de Órganos Artificiales, Biomateriales e Ingeniería de Tejidos; 2013

SLABO Sociedad Latinoamericana de Órganos Artificiales, Biomateriales e Ingeniería de Tejidos

CRYOPRESERVATION OF RAT NEURONAL CELLS BY SLOW COOLING METHODOLOGY Juan de Paz Leonardo1,2, Robert M.Celeste1, Berardi Florencia1,Balaban Cecilia L.1, Graf Daniel2, Gazzin Silvia3, Tiribelli Claudio3, Rodriguez Joaquín V.1 1Centro Binacional (Argentina-Italia) de Investigaciones en Criobiología Clínica y Aplicada (CAIC), UNR.2 Servicio de Electrónica y Óptica, FCByF, UNR, Argentina. 3Fondazione Italiana Fegato (Italian Liver Foundation), Centro Studi Fegato, Trieste, Italy. * jrodrig@fbioyf.unr.edu.ar ABSTRACT Slow cooling methodology has been widely applied for cryopreservation of multiple mammalian tissues. Nevertheless, relatively low success has been achieved in the case of nervous tissue. Usually applied methodologies are adapted and designed based on observations made on unrelated cell types. Our work deals with the optimization of a slow cooling methodology with spontaneous ice nucleation applied to primary rat neuronal cells. Parameters such as cooling rate and liquid nitrogen plunge temperature were evaluated in relation with post-rewarming viability and viable cell yield. In-culture studies were also performed and the general development of the culture was monitored for fourteen days. Results suggest that, in our presented protocol, most favorable conditions were achieved with cells cooled at 3.1 ± 0.5 ºC/min until sample temperature reaches ~ -50ºC and then plunged in liquid nitrogen for storage.