Enrichment of human heterokaryons by ficoll gradient for complementation analysis of iduronate sulfatase deficiency |
| |
Authors: | Lorraine R Eisenberg Barbara R Migeon |
| |
Institution: | (1) Department of Pediatrics, Johns Hopkins University School of Medicine, 21205 Baltimore, Maryland |
| |
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. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |