Embryo‐fetal erythroid cell selection from celomic fluid allows earlier prenatal diagnosis of hemoglobinopathies

Abstract

Celocentesis can be a reliable procedure for earlier prenatal diagnosis of fetal monogenic diseases.

Identification of embryo–fetal cells in celomic fluid using morphological and short‐tandem repeats analysis

“…The diagnostic procedure for contaminated CF samples requires selection of pure fetal cells of type 2/3/5 and 8 to remove contamination and makes celocentesis a reliable procedure for early prenatal diagnosis of Mendelian disorders and chromosomal analysis by standard methodology or microarray comparative genomic hybridization (CGH) . Our experimental results, described in this manuscript, and the 353 prenatal diagnosis performed by celocentesis for hemoglobinopaties, without any diagnostic errors, allow us to propose the procedure of the fetal cell selection to solve the contamination in the CF samples. Celocentesis remains an attractive technique for early prenatal diagnosis of genetic disorders as demonstrated by a growing demand from couples at risk for hemoglobinopathies coming from all regions of Italy.…”

“…Their source is likely to be because of contamination from tissues crossed during the sampling; however, their possible origin from maternal–fetal celomic circulation cannot be excluded. Four types of embryo–fetal cells have been identified: one type could be fibroblast cells, another with pseudopodia could be neuronal cell precursors and two others are megaloblasts (CD 71+/CD 45−) previously reported …”

“…The robustness and reproducibility of this methodology to classify these cells in relation to their morphology were demonstrated by the correct association of their morphology to their origin in all pooled cells analyzed. As recently reported, 302 CF were examined for prenatal diagnosis of hemoglobinopathies by applying a flow work which required microscopic assessment and cell selection by micromanipulation of fetal cells based on their morphology. The results obtained by celocentesis in this clinical diagnostic trial were all subsequently confirmed with amniocentesis or CVS.…”

“…The present study has demonstrated the feasibility to distinguish between embryo–fetal and maternal cells in CF. It is now possible to isolate embryo–fetal cells after positive selection using antibodies or individually by micromanipulator picking‐up making it possible to solve concerns about maternal DNA contamination . The third problem relates to uncertainty of the safety of the technique.…”

“…Four types of embryo-fetal cells have been identified: one type could be fibroblast cells, another with pseudopodia could be neuronal cell precursors and two others are megaloblasts (CD 71+/CD 45À) previously reported. 26,27 Hematopoiesis in the embryo starts in the 2nd or 3rd week of gestation when yolk sac blood islands develop clones of primitive nucleated erythroblasts or megaloblasts. [28][29][30] Detailed analysis of these embryonic cells revealed a prevalent megaloblast pattern with CD71+/CD45À membrane expression, consistent with embryonic erythroid precursors.…”