The use ofin vitro expanded erythroid cells in a model system for the isolation of fetal cells from maternal blood
| Autor: | Juriy W. Wladimiroff, Helen Brandenburg, Hartmut Beug, Peter A. In 't Veld, Mieke W. J. C. Jansen, Hajo I. J. Wildschut, Marieke von Lindern |
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| Rok vydání: | 1999 |
| Předmět: |
Fetus
Pathology medicine.medical_specialty Magnetic-activated cell sorting medicine.diagnostic_test Obstetrics and Gynecology Nucleated Red Blood Cell Biology Umbilical cord Flow cytometry Andrology Red blood cell medicine.anatomical_structure Erythroblast medicine Genetics (clinical) Fluorescence in situ hybridization |
| Zdroj: | Prenatal Diagnosis. 19:323-329 |
| ISSN: | 1097-0223 0197-3851 |
| Popis: | The development of a non-invasive prenatal diagnostic test using fetal nucleated red blood cells (NRBCs) isolated from the maternal circulation is hampered by the low frequency of these cells in maternal blood, requiring extensive enrichment procedures before any analytical procedure can be performed. In order to improve and simplify these procedures, we have used in vitro expanded erythroid cells derived from male umbilical cord blood in a model system for the isolation of fetal NRBCs from maternal blood. Erythroblast cells were expanded in vitro to high cell numbers and were immunophenotypically identical to fetal NRBCs isolated from maternal blood. Magnetic activated cell sorting (MACS) isolation procedures were optimized using in vitro expanded male NRBCs diluted up to 1 in 400,000 with female peripheral blood mononucleated cells. The number of recovered male cells was determined using two-colour fluorescence in situ hybridization with X and Y chromosomal probes. Using this model system, an NRBC isolation technique is described. It is based on a one-step MACS enrichment protocol for CD71 positive cells, which showed a significant (Wilcoxon signed ranks test, p |
| Databáze: | OpenAIRE |
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