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Journal of the Society for Gynecologic Investigation
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The Effect of the Elapsed Time Between Blood Draw and Processing on the Recovery of Fetal Cells From Maternal Blood

K. A. Dukes, PhD

DM-STAT Inc., Medford, Massachusetts; Department of Biostatistics, Boston University School of Public Health, Boston, Massachusetts; Department of Immunology, Baylor College of Medicine, Houston, Texas; Division of Genetics, Departments of Pediatrics, Obstetrics, and Gynecology, Tufts University School of Medicine, Boston, Massachusetts; Departments of Obstetrics and Gynecology, and Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas; Department of Obstetrics and Gynecology, University of Basel, Basel, Switzerland; Department of Obstetrics and Gynecology, Drexel University College of Medicine, Philadelphia, Pennsylvania; Kim.Dukes{at}dmstat.com

L. M. Sullivan, PhD

D. Lewis, PhD

K. L. Johnson, PhD

D. W. Bianchi, MD

J. L. Simpson, MD

W. Holzgreve, MD

S. Hahn, PhD

F. Z. Bischoff, PhD

L. G. Jackson, MD

DM-STAT Inc., Medford, Massachusetts; Department of Biostatistics, Boston University School of Public Health, Boston, Massachusetts; Department of Immunology, Baylor College of Medicine, Houston, Texas; Division of Genetics, Departments of Pediatrics, Obstetrics, and Gynecology, Tufts University School of Medicine, Boston, Massachusetts; Departments of Obstetrics and Gynecology, and Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas; Department of Obstetrics and Gynecology, University of Basel, Basel, Switzerland; Department of Obstetrics and Gynecology, Drexel University College of Medicine, Philadelphia, Pennsylvania

Objective: To test the hypothesis that a delay in initial fetal cell enrichment processing of maternal blood samples (defined as the time between blood draw and the initial density gradient centrifugation step) compromises the ability to recover fetal cells, we performed a randomized comparison of immediate (within 4 hours of draw) versus delayed (between 18-24 hours of draw) processing.

Methods: Four centers participated: two centers utilized flow cytometry (FLOW), and two centers utilized magnetic-activated cell sorting (MACS) techniques. Each center collected 34 samples. The outcome was the percentage of gamma positive ({gamma}+) cells for FLOW or the number of nucleated red blood cells (NRBCs) for MACS, found in the final enriched cell population. Both outcomes reflect cell properties that are potentially fetal in origin, thus making them representative of the ability to recover fetal cells.

Results: Our results did not support our hypothesis that delay in processing compromises fetal cell recovery. Instead, in MACS processing, we observed an increase in recovered NRBCs when blood sample processing was delayed compared with immediate processing. There was no significant difference in {gamma}+ cells with FLOW.

Conclusion: Time-related changes in the density of target cells, perhaps associated with their progress towards apoptosis during the delay period, may result in increased intact fetal cells with the study methods utilized.

Key Words: Fetal cells in maternal blood • noninvasive prenatal diagnosis • fetal cell recovery

Journal of the Society for Gynecologic Investigation, Vol. 11, No. 3, 154-165 (2004)
DOI: 10.1016/j.jsgi.2003.09.005
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