Fortuin J J, Kleijer W J
Clin Chim Acta. 1978 Jan 2;82(1-2):79-83. doi: 10.1016/0009-8981(78)90028-1.
Normal fibroblasts and amniotic fluid cells, and cells from patients with Mucopolysaccharidosis type I and II, were cultured in the presence of 35SO4. After harvesting by trypsinization, the radioactivity was recovered from the intracellular and pericellular pool of glycosaminoglycans. The amount of incorporated radioactivity in these respective pools was 1 : 2.3 in normal fibroblasts and 1 : 7.2 in control amniotic fluid cells. Incorporation in the pericellular pool was not elevated in cells from patients with Mucopolysaccharidosis type I and II, in contrast to incorporation in the intracellular pool. Studies on different methods of harvesting showed that reliable prenatal analysis can be performed only if the pericellular pool is removed by trypsinization. Amniotic fluid cells from a pregnancy carrying a fetus affected with Hurler's disease revealed the expected increased level of 35SO4 incorporation if the cells were trypsinized, but the intracellular accumulation of glycosaminoglycans was obscured by the pericellular pool if the cells were harvested by scraping.
将正常成纤维细胞、羊水细胞以及患有I型和II型黏多糖贮积症患者的细胞置于含有35SO4的环境中培养。通过胰蛋白酶消化法收获细胞后,从细胞内和细胞周围的糖胺聚糖池中回收放射性物质。在这些相应的池中,正常成纤维细胞中掺入的放射性物质的量之比为1:2.3,对照羊水细胞中为1:7.2。与细胞内池中的掺入情况相反,I型和II型黏多糖贮积症患者细胞的细胞周围池中的掺入并未增加。对不同收获方法的研究表明,只有通过胰蛋白酶消化法去除细胞周围池,才能进行可靠的产前分析。如果对患有Hurler病胎儿的妊娠羊水细胞进行胰蛋白酶消化,会发现35SO4掺入水平如预期增加,但如果通过刮取收获细胞,细胞周围池会掩盖糖胺聚糖的细胞内积累。