Comparison of methods for erythroblast selection: Application to selecting fetal erythroblasts from maternal blood

• Published in: Cytometry (New York, N.Y.) Vol. 45; no. 4; pp. 267 - 276
• Main Authors: Collarini, Ellen J., Cain, Carol A., Gammon, Dawn, Harriman, Bill, Magee, Kerry, Du, Gehua, Schueler, Paula A., Mahoney, Walt C.
• Format: Journal Article
• Language: English
• Published: New York John Wiley & Sons, Inc 01-12-2001
• Subjects: Acetazolamide - pharmacology; Centrifugation, Density Gradient - methods; erythroblasts; Erythroblasts - cytology; Erythroblasts - drug effects; Female; Fetal Blood - cytology; Fetal Blood - drug effects; fetal cells in maternal blood; Fetomaternal Transfusion; Gestational Age; Hemolysis; Humans; immunomagnetic selection; Immunomagnetic Separation - methods; In Vitro Techniques; Pregnancy - blood; prenatal diagnosis; Prenatal Diagnosis - methods; Receptors, Transferrin - immunology
• ISSN: 0196-4763; 1097-0320
• DOI: 10.1002/1097-0320(20011201)45:4<267::AID-CYTO10023>3.0.CO;2-D

Background
Many methods have been employed to obtain fetal cells from maternal blood for prenatal diagnostics, but there has been little work done that compares the efficacy of different methods. This study presents a comparison of two commonly used methods for selecting erythroblasts with selection directly from whole blood.
Methods
Erythroblasts were isolated from maternal blood by either differential lysis or density separation, followed by selection with an antibody to the transferrin receptor. These methods were compared with antibody selection directly from whole blood. The total yield of erythroblasts was determined for each method.
Results
Red cell lysis is not recommended because the lysis step cannot be well controlled. Density separation followed by antibody selection works well. However, a faster and simpler method, antibody selection directly from whole blood using Immunicon Ferrofluid and magnetic separators, works as well and has the potential to yield even more cells.
Conclusions
Considering the need for a simple and quick method for selecting fetal cells from maternal blood, we suggest selection directly from whole blood. Cytometry 45:267–276, 2001. © 2001 Wiley‐Liss, Inc.