Effects of cryopreservation and hypothermic storage on cell viability and enzyme activity in recombinant encapsulated cells overexpressing alpha-L-iduronidase.
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.
Mayer
Fabiana Quoos
FQ
Centro de Terapia Gênica, Centro de Pesquisas, Hospital de Clínicas de Porto Alegre, Rua: Ramiro Barcelos 2350, Porto Alegre, RS, Brazil.
Baldo
Guilherme
G
de Carvalho
Talita Giacomet
TG
Lagranha
Valeska Lizzi
VL
Giugliani
Roberto
R
Matte
Ursula
U
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
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D007068:Iduronidase