An examination of different Percoll density gradients and magnetic activated cell sorting (MACS) for the enrichment of fetal erythroblasts from maternal blood.
OBJECTIVE
To increase the yield of fetal erythroblasts from the blood of pregnant women by optimising and simplifying the enrichment procedure.


METHODS
By using an experimental system whereby cord blood was mixed with normal adult blood, we evaluated the recovery of fetal erythroblasts using different single or double percoll density gradients with or without subsequent enrichment by Magnetic Activated Cell Sorting (MACS).


RESULTS
The recovery of erythroblasts on single density gradients increased from 3%-68% of the input erythroblasts with an increase in the density of percoll from 1075 g/l-1098 g/l. Although the use of a double density gradient did eliminate most lymphocytes, it was accompanied by a significant loss of erythroblasts. The combination with MACS enrichment, however, lead to a almost 200 fold increase in the purity of erythroblasts.


CONCLUSIONS
Erythroblasts are preferentially retained by higher density gradients. To enhance their purity a subsequent more specific enrichment step, such as MACS is advisable.
Smits
G
G
Department of Obstetrics and Gynaecology, University of Basel, Switzerland.
Holzgreve
W
W
Hahn
S
S
D001772:Blood Cell Count; D002499:Centrifugation, Density Gradient / Q000379:methods*; D004900:Erythroblasts / Q000737:chemistry*; D005260:Female; D005312:Fetal Blood / Q000166:cytology; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D011205:Povidone / Q000737:chemistry; D011247:Pregnancy / Q000097:blood*; D011296:Prenatal Diagnosis / Q000379:methods*; D012822:Silicon Dioxide
C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone