Enrichment of Fetal Nucleated Red Blood CellsCirculating in the Peripheral Blood of Pregnant Womenfor the Purpose of Non-invasive Prenatal Diagnosis
Authors:
I. Hromadníková 1; L. Sedláčková 2; M. Mrštinová 3; K. Štechová 1; KaramanovS. 5; J. Kofer 5; M. Macek 4
Authors‘ workplace:
II. dětská klinika 2. LF UK, FN Motol, Praha, přednosta doc. MUDr. J. Vavřinec, CSc. 2 Laboratoř imunologie, KlinLab, Praha 3 Gynek. -porod. klinika FN Motol, Praha, přednosta doc. MUDr. K. Citterbart, CSc. 4 Oddělení lékařské genetiky, FN Motol, Praha, p
1
Published in:
Ceska Gynekol 2000; (1): 33-37
Category:
Overview
Objective:
Enrichment of nucleated red blood cells (NRBCs) from maternal blood for non-invasiveprenatal diagnosis.Design: Pilot study.Setting: 2 nd Clinic of Paediatrics, University Hospital Motol, Prague, Czech Republic.Methods: Mononuclear cells were isolated from 13 - 28 ml of peripheral maternal blood between 13and 37 weeks of gestation. Leukocytes from maternal peripheral blood were depleted from mono-nuclear cells by treatment with anti-CD14 and anti-CD45 microbeads and high-gradient magneticcell separation (MACS) on VarioMACS. NRBCs were sorted from CD14 - /CD45 - fraction by positiveselection using anti-CD71 microbeads on MiniMACS. All sorting steps were analysed by three-co-lour cytometric analysis with FACScan flow cytometer.Results: In 68 out of 78 pregnant woman (87 %) NRBCs were found in range 2x10 5 - 1,02 x 10 6.NRBC were enriched with an average enrichment rate of 138-fold ranging from 4 - 526 fold. In ourcohort of pregnant woman the number of isolated NRBCs was individual. We identified NRBCsfrom the 13 th week of gestation.Conclusion: The aim of the study is to establish and standardise the method of enrichment ofNRBCs from maternal blood samples and verify the applicability of this alternative source fornon-invasive prenatal diagnosis.
Key words:
nucleated red blood cells, high-gradient magnetic cell separation (MACS), flow cyto-metry (FACS), prenatal diagnosis, fetal cells.
Labels
Paediatric gynaecology Gynaecology and obstetrics Reproduction medicineArticle was published in
Czech Gynaecology
2000 Issue 1
Most read in this issue
- Laparoscopic Excision of Endometriosis of theRectovaginal Septum by Diomed Laser
- Hormonal Substitution Treatment in Women Operatedon Account of Endometriosis and Adenomyosis:Prospective Follow-up Study - Part I
- Paracervical Anaesthesia in Hysteroscopy andTranscervical Surgery
- Statistics for Medical Doctors - Part I