PMID- 10420047
OWN - NLM
STAT- MEDLINE
DCOM- 19990902
LR  - 20171101
IS  - 1015-3837 (Print)
IS  - 1015-3837 (Linking)
VI  - 14
IP  - 4
DP  - 1999 Jul-Aug
TI  - Improvement of fetal cell recovery from maternal blood: suitable density gradient 
      for FACS separation.
PG  - 229-33
AB  - OBJECTIVE: To improve the recovery of fetal nucleated erythrocytes (NRBCs) from 
      maternal blood for noninvasive prenatal genetic diagnosis. METHODS: Blood samples 
      were obtained from 10 women at 8-22 weeks of gestation. Samples were split and 
      mononuclear cells were isolated using 1.083 and 1.090 g/ml of Percoll solution. 
      Flow sorting with antibody to fetal hemoglobin and fluorescence in situ 
      hybridization (FISH) analysis were used to evaluate the number of fetal cells 
      recovered. RESULTS: In samples separated with the 1.090 density gradient, the 
      yield of true gamma-hemoglobin-positive cells (median 21.0, range 2.2-303.8) was 
      1.9 times higher than that in the 1.083 density (median 11.1, range 1.1-87.5), 
      although it took 2. 1-fold longer time to flow sort the gamma-hemoglobin-positive 
      cells. In 7 out of 10 cases, the number of gamma-hemoglobin-positive cells 
      recovered from the 1.090 density gradient was 3 times or greater than that from 
      1.083 gradient. After FISH analysis, we detected a median of 13.3 (range 
      2.2-98.8) fetal NRBCs per 10-ml maternal blood in the 1.090 density gradient, 
      whereas a median of 11.0 fetal NRBCs were detected in the 1.083 gradient (range 
      1.1-35.0). The number of fetal NRBCs in the 1.090 density was significantly 
      higher than that in the 1.083. CONCLUSION: Increased Percoll density results in 
      improved fetal cell recovery in fresh posttermination maternal samples. The 
      increased yield of fetal cells using this gradient may permit better noninvasive 
      detection of fetal chromosome as well as DNA abnormalities in maternal blood.
FAU - Sekizawa, A
AU  - Sekizawa A
AD  - Division of Genetics, Departments of Pediatrics, Obstetrics and Gynecology, New 
      England Medical Center, Tufts University School of Medicine, Boston, Mass. 02111, 
      USA.
FAU - Farina, A
AU  - Farina A
FAU - Zhen, D K
AU  - Zhen DK
FAU - Wang, J Y
AU  - Wang JY
FAU - Falco, V M
AU  - Falco VM
FAU - Elmes, S
AU  - Elmes S
FAU - Bianchi, D W
AU  - Bianchi DW
LA  - eng
GR  - N01-HD-4-3204/HD/NICHD NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Switzerland
TA  - Fetal Diagn Ther
JT  - Fetal diagnosis and therapy
JID - 9107463
RN  - 9007-49-2 (DNA)
RN  - 9034-63-3 (Fetal Hemoglobin)
SB  - IM
MH  - Aneuploidy
MH  - Cell Separation/*methods
MH  - DNA/blood/genetics
MH  - Erythroblasts/cytology/metabolism
MH  - Female
MH  - Fetal Blood/*cytology/metabolism
MH  - Fetal Diseases/blood/diagnosis/genetics
MH  - Fetal Hemoglobin/metabolism
MH  - Flow Cytometry/*methods
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Karyotyping
MH  - Male
MH  - Pregnancy/*blood
MH  - Prenatal Diagnosis/*methods
EDAT- 1999/07/27 00:00
MHDA- 1999/07/27 00:01
CRDT- 1999/07/27 00:00
PHST- 1999/07/27 00:00 [pubmed]
PHST- 1999/07/27 00:01 [medline]
PHST- 1999/07/27 00:00 [entrez]
AID - 20927 [pii]
AID - 10.1159/000020927 [doi]
PST - ppublish
SO  - Fetal Diagn Ther. 1999 Jul-Aug;14(4):229-33. doi: 10.1159/000020927.