Cell sorting and microchemistry of cultured human fibroblasts: applications in genetics and aging research.
Cell sorting is a way to isolate viable subpopulations of cells present in a mixture. The drawback of the isolation method is the shortage of material for subsequent biochemical determinations. We have employed a combination of (ultra-) microchemistry and cell sorting to overcome this problem. The methods enable determinations of protein and several enzyme activities on triton extracts of 5,000-10,000 sorted cells. In addition, using ultramicromethods we could determine enzyme activity in single sorted cells. This combination of methods is used for clinical genetic studies on heterozygote detection in Fabry's disease, an X-linked genetic disease. Moreover, microchemistry is used to study enzyme activities in sorted autofluorescent "aged" fibroblasts.
D000375:Aging; D002478:Cells, Cultured; D000795:Fabry Disease / Q000235:genetics*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D005434:Flow Cytometry / Q000379:methods; D006580:Genetic Carrier Screening; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D006801:Humans; D011506:Proteins / Q000032:analysis; D012867:Skin / Q000166:cytology; D012879:Skin Physiological Phenomena; D001616:beta-Galactosidase / Q000378:metabolism
D011506:Proteins; D005954:Glucosephosphate Dehydrogenase; D001616:beta-Galactosidase