Improved Cultivation of Amniotic Fluid Chorial and Placental Cells for Prenatal Diagnosis

Overview

Long-term follow up amniotic fluid cell cultures brought evidence that in amniotic fluid three types of cellsexist, with regard to the rapidity of adherence to the surface of tissue culture flasks. In addition to well knowncells, adhering within 1 - 5 days of cultivation, the authors revealed cells rapidly adhering to the surface of transportculture vessels within the range of 1 - 5 hours, during transport of aspirated amniotic fluid to the laboratory andthe late adhering cells, which adhere to the culture vessel after 5 - 12 days of cultivation. Rapidly and late adheringamniotic fluid cells are lost by the current method of amniocyte cultivation. Both types of cells provide successfulparallel cultures, with 100% cultivation and diagnostic success from the 13th week to the end of gestation.Combination of cultivation of all types of amniotic fluid cells with the cultivation of microfragments of chorionicor placental villi ensures a 100% diagnostic success from the end of Ist to the end of the IIIrd trimester. The time,necessary for obtaining large colonies in vitro and to perform first subcultivation is identical in all types of amnioticfluid cells as well as in chorionic or placental villi cells. In all these cells, a tendency of slower growth after the 30thweek of gestation is apparent. The described methods provide a reliable prenatal diagnosis, the greatest possibilityto detect chromosomal mosaicism and the chance to finalize results of prenatal diagnosis in the early phase of thesecond trimeter and to ensure cytogenetic analysis of high risk pregnancies with IUGR or severe congenitalanomalies in the IIIrd trimester. This method offers a diagnostic background for new meth ods of biochemical andultrasound screening in the 9th - 14th week of gestation.

Key words:
prenatal diagnosis, rapidly adhering amniotic fluid cells, slowly adhering amniotic fluid cells, 1st,