Time-lapse cinemicrography has been used to study cell behaviour and movement in long- and short-term amniotic fluid cultures from various types of fetus. These included several with a neural tube defect (9 anencephalics, 5 spina bifidas, 2 encephaloceles), 1 with an abdominal wall lesion (omphalocele) and 9 controls with no open lesion. In short-term (less than 3 days) amniotic fluid cultures from normal fetuses, non-adherent squamous cells dominate, but there is a small population of cells which, in the longer term (approximately 4 weeks), will form colonies of epithelioid and fibroblastic cells. In addition, there is a further group, called 'AF' by Hoehn et al. (1974), which have the ability to form syncitial strands and are often multinucleated; these cells seem to be trophoblastic in origin. In contrast to the controls, short-term anencephalic cultures contained many adherent cells; these were mainly latex-particle-phagocytosing macrophages and neural cells. In long-term cultures, the neural cells dominate and differentiate into a range of recognizable forms whose type and behaviour seem to depend on cell density, time in culture and extent of cell-cell contacts. In the spina bifida and encephalocele cultures, there were initially far fewer adherent cells than in the anencephalic ones, but, after approximately 2 weeks in vitro, a range of neural cell types could be recognized in addition to those seen in the controls. In the context of prenatal diagnosis, the presence of neural cells certainly indicates that the fetus has a NTD, but the difficulty of standardizing culture conditions implies that any differential diagnosis on the basis or morphology and movement will be difficult. If, as seems likely, the AF cells of Hoehn et al. (1974) are indeed trophoblastic, amniotic fluids from the second trimester may provide a useful source of these cells.

Amniotic fluid cells from normal and abnormal fetuses (neural tube defects and abdominal wall lesions) were examined uncultured and after short-term culture for macrophages. Morphology, adherence, phagocytosis, presence of Fc receptors and non-specific esterase were studied. The existence of macrophages in normal and abnormal fluids was confirmed, although the percentage and the absolute numbers varied greatly from specimen to specimen. The most marked increases in total macrophages were in cases of anencephaly. The significance of these macrophages and their value in prenatal diagnosis are discussed.

The cell morphology of long-term cultures of amniotic fluid cells from 10 fetuses with a neural tube defect (NTD) and three with omphalocele was examined and compared to 30 long-term cultures of normal amniotic fluids as well as a long-term culture of human fetal brain. Cultures from the amniotic fluids of the fetuses with NTD and omphalocele showed cells with the same general characteristics as normal amniotic fluid cells. However, the cultures of amniotic fluid cells from NTD pregnancies had an additional cell type also seen in fetal brain culture. This was a neuroblast-like cell, with small rounded refractile morphology and long branching processes forming clusters of varying sizes which lay on top of large flat cells. These neuroblast-like cells diminished in number with time in culture and were not present in subcultures. Their possible neuronal origin is discussed.

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Amniotic fluid cells have been widely used in prenatal diagnosis; however, there is great heterogeneity of the cells and their origin. In this study we analyze the karyotype and release of human chorionic gonadotropin (hCG), human chorionic somatomammotropin (hCS), free estriol (E 3), prolactin (PRL) and progesterone (P) of amniotic fluid cells from primary cultures of six normal and two anencephalic fetuses. In all the amniotic fluid samples there was release of hCG; in one amniotic fluid, in which several tetraploid colonies were found. PRL and P were also released. The heterogeneity of amniotic fluid cell morphology and their hormone release in culture was confirmed. The presence of hormones like hCG supports the trophoblastic origin of some amniotic fluid cells from normal and anencephalic fetuses. Other hormones, such as PRL and P could be used in the differential diagnosis between the karyotype of fetal membranes and the true fetal karyotype. Amniotic fluid cell cultures used in prenatal diagnosis yielded second trimester placental cells without any elaborate methods that could be used as cell models for hormone studies.