title: Effects of cryopreservation and hypothermic storage on cell viability and enzyme activity in recombinant encapsulated cells overexpressing alpha-L-iduronidase.
abstract: Here, we show the effects of cryopreservation and hypothermic storage upon cell viability and enzyme release in alginate beads containing baby hamster kidney cells overexpressing alpha-L-iduronidase (IDUA), the enzyme deficient in mucopolysaccharidosis type I. In addition, we compared two different concentrations of alginate gel (1% and 1.5%) in respect to enzyme release from the beads and their shape and integrity. Our results indicate that in both alginate concentrations, the enzyme is released in lower amounts compared with nonencapsulated cells. Alginate 1% beads presented increased levels of IDUA release, although this group presented more deformities when compared with alginate 1.5% beads. Importantly, both encapsulated groups presented higher cell viability after long cryopreservation period and hypothermic storage. In addition, alginate 1.5% beads presented higher enzyme release after freezing protocols. Taken together, our findings suggest a benefic effect of alginate upon cell viability and functionality. These results may have important application for treatment of both genetic and nongenetic diseases using microencapsulation-based artificial organs.
authors.affiliation: Centro de Terapia Gênica, Centro de Pesquisas, Hospital de Clínicas de Porto Alegre, Rua: Ramiro Barcelos 2350, Porto Alegre, RS, Brazil.
authors.forename: Fabiana Quoos
authors.initials: FQ
authors.lastname: Mayer
authors.forename: Guilherme
authors.initials: G
authors.lastname: Baldo
authors.forename: Talita Giacomet
authors.initials: TG
authors.lastname: de Carvalho
authors.forename: Valeska Lizzi
authors.initials: VL
authors.lastname: Lagranha
authors.forename: Roberto
authors.initials: R
authors.lastname: Giugliani
authors.forename: Ursula
authors.initials: U
authors.lastname: Matte
mesh_terms: D000464:Alginates / Q000737:chemistry; D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D018914:Cells, Immobilized / Q000201:enzymology; D003080:Cold Temperature; D006224:Cricetinae; D015925:Cryopreservation; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D007068:Iduronidase / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism*; D007668:Kidney / Q000166:cytology*; D008059:Mucopolysaccharidosis I / Q000201:enzymology; D015854:Up-Regulation
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D007068:Iduronidase