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Flow sorting of fetal erythroblasts using intracytoplasmic anti‐fetal haemoglobin: Preliminary observations on maternal samples
Author(s) -
Zheng YunLing,
Demaria Maryann,
Zhen Dongkai,
Vadnais Theresa J.,
Bianchi Diana W.
Publication year - 1995
Publication title -
prenatal diagnosis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.956
H-Index - 97
eISSN - 1097-0223
pISSN - 0197-3851
DOI - 10.1002/pd.1970151004
Subject(s) - fetus , fluorescence in situ hybridization , biology , microbiology and biotechnology , flow cytometry , staining , fluorescein isothiocyanate , monoclonal antibody , andrology , pathology , antibody , immunology , pregnancy , chromosome , medicine , genetics , fluorescence , gene , physics , quantum mechanics
Abstract Monoclonal antibody to fetal haemoglobin ( a 2 γy 2 ) has been proposed as a fetal‐specific reagent. We developed an intracellular staining protocol that combines fluorescein isothiocyanate or phycoerythrin conjugated anti‐γ with the DNA binding dye Hoechst 33342 to identify and flow sort fetal erythroblasts from maternal blood. Our preliminary observations on anti‐γ‐positive cells sorted from four different pregnant women are described here, using fluorescence in situ hybridization (FISH) with chromosome‐specific probes to identify fetal cells. Our data demonstrate that far fewer candidate fetal cells are sorted with this protocol than by current cell surface staining methods that employ the monoclonal antibody CD71. This results in increased fetal cell sorting purities. With this protocol, standard FISH techniques require modification due to the rigorous fixation with 4 per cent paraformaldehyde. Our initial data indicate the promise of this approach.