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Rapid tissue culture and microbiochemical methods for analyzing colonially grown fibroblasts from normal, Lesch‐Nyhan and Tay‐Sachs patients and amniotic fluid cells
Author(s) -
Richardson B. J.,
Cox D. M.
Publication year - 1973
Publication title -
clinical genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 102
eISSN - 1399-0004
pISSN - 0009-9163
DOI - 10.1111/j.1399-0004.1973.tb01163.x
Subject(s) - amniocentesis , hypoxanthine , amniotic fluid , lesch–nyhan syndrome , prenatal diagnosis , hexosaminidase , tay sachs disease , cell culture , biology , fetus , enzyme , microbiology and biotechnology , gangliosidosis , andrology , medicine , hypoxanthine guanine phosphoribosyltransferase , pregnancy , biochemistry , genetics , disease , gene , mutant
A means of reducing the time interval between amniocentesis and biochemical analysis for the prenatal diagnosis of biochemical diseases is presented. Fibroblasts from nomal individuals, patients with Lesch‐Nyhan syndrome and patients with Tay‐Sachs disease, and normal amniotic fluid cells, were grown in isolation in MicroTest II tissue culture plates. Hypoxanthine‐guanine phosphoribosyl transferase (HGPRTase) and hexosaminidase A were studied in independently derived colonies arising after a median culture period of 14 days. Colony six ranged from 2,000 to 8,000 cells. HGPRTase activity, expressed as the ratio of hypoxanthine‐C14 to adenine‐113 uptake, was assayed using differential scintillation spectrometry. Hexosaminidase A was investigated by micro‐electrophoresis. Discrimination of enzyme deficient from non‐deficient colonies was accomplished in each case and both enzymes were detectable in amniotic fluid cell colonies. The potential of the method for prenatal diagnosis of the two diseases studied and of biochemical diseases in general is discussed.