Subzero nonfreezing storage of rat hepatocytes using UW solution and 1.4-butanediol.

II- functional testing on rewarming and gene expression of urea cycle enzymes 

Autores: Guibert Edgardo E, Almada Luciana L, Mamprin María Eugenia, Bellarosa Cristina, Pizarro María Dolores, Tiribelli Claudio, Rodríguez Joaquín V

Resumen

In the present study we have analyzed the viability and metabolic competence of isolated rat hepatocytes subjected first, to subzero nonfreezing storage (up to 120 h at -4º C) in modified University of Wisconsin (UW) solution with 8% 1,4-butanediol, and then to a normothermic rewarming step (KHR media, 37º C, up to 120 min, carbogen atmosphere). Results were compared with hepatocytes stored up to 120 h at 0º C in modified UW solution and with freshly isolated hepatic cells. We have found that only cell suspensions stored in subzero nonfreezing conditions were able to finish the rewarming period with a viability comparable with the control group. Also, we have investigated the enzyme activities and the relative expression at messenger RNAs levels of two of the Urea cycle (UC) enzymes: Carbamyl phosphate synthetase I (CPSI) and ornithine transcarbamylase (OTC), during 60 min of rewarming. Results were compared with the ammonium removal efficiency of the three groups. In conclusion: These data indicated that hepatocytes preserved under cold or subzero conditions up to 120 h followed by 60 min of rewarming, maintain UC enzymes at levels similar to freshly isolated hepatocytes, allowing their use in bioartificial liver devices.

Palabras clave: Subzero cold storage gene expression urea cycle enzymes.

2011-07-13   |   439 visitas   |   Evalua este artículo 0 valoraciones

Vol. 8 Núm.2. Abril-Junio 2009 Pags. 129-133 Ann Hepatol 2009; 8(2)