Optimization of nucleated red blood cell (NRBC) recovery from maternal blood collected using both layers of a double density gradient

Belén Prieto, Rebeca Alonso, Ana Paz, Mercedes Cándenas, Rafael Venta, Jack H. Ladenson, Francisco V. Álvarez

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

The isolation of fetal nucleated red blood cells (NRBC) from maternal blood represents a promising approach to non-invasive prenatal diagnosis. However, the number of fetal NRBC in maternal circulation is quite low and therefore difficult to isolate. An enrichment procedure in which both layers from a double density 1.077/1.107 g/ml gradient are collected was optimized, followed by MACS selection using non-commercial monoclonal antibodies. The influence of the delay in processing maternal blood on the NRBC distribution in both interfaces of the gradient was also studied in cord blood and peripheral maternal blood samples. A significant increase in the number of NRBC isolated from maternal blood was achieved by collecting both layers of the double density gradient compared with the previous protocol in which only the lower layer was recovered. Cord blood samples showed significant differences in the number of NRBC recovered when processed at 24 instead of within 3 h. This effect was also observed in the number of NRBC collected only from the upper layer of peripheral maternal blood samples. Therefore, in order to minimize the target cell losses, it is advisable to process the maternal blood samples as soon as possible.

Original languageEnglish
Pages (from-to)187-193
Number of pages7
JournalPrenatal Diagnosis
Volume21
Issue number3
DOIs
StatePublished - Apr 17 2001

Keywords

  • Double density gradient
  • Fetal cell enrichment
  • Fetal nucleated red blood cells

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