title: Enrichment of human heterokaryons by Ficoll gradient for complementation analysis of iduronate sulfatase deficiency.
abstract: Ficoll gradients have been used to enrich for heterokaryons in cultures of human skin fibroblasts following polyethylene glycol (PEG) induced fusion. These gradients provide a simple and consistent method for obtaining populations of multinucleated cells, at least twofold greater than those resulting from fusion alone. Formation of glucose-6-phosphate dehydrogenase (G6PD) heteropolymers has been used as a functional assay for the presence of heterokaryons. Analysis of cell populations enriched for multinucleated cells has revealed complementation leading to iduronate sulfatase activity in heterokaryons derived from iduronate sulfatase-deficient fibroblasts expressing the Hunter and multiple sulfatase-deficiency mutations.
mesh_terms: D002499:Centrifugation, Density Gradient / Q000379:methods*; D005362:Ficoll; D005816:Genetic Complementation Test; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D006801:Humans; D006822:Hybrid Cells / Q000201:enzymology; D007066:Iduronate Sulfatase / Q000235:genetics*; D016532:Mucopolysaccharidosis II / Q000235:genetics; D010641:Phenotype; D012867:Skin / Q000166:cytology / Q000201:enzymology; D013429:Sulfatases / Q000235:genetics*
chemical_list: D005362:Ficoll; D005954:Glucosephosphate Dehydrogenase; D013429:Sulfatases; D007066:Iduronate Sulfatase