How useful is the in vitro expansion of fetal CD34+ progenitor cells from maternal blood samples for diagnostic purposes?
Fetal cells present in the maternal circulation are a potential source of fetal DNA that can be used for the development of a prenatal diagnostic test. Since their numbers are very low, amplification of fetal cells has been discussed for a long time. So far, most studies have focused on culturing fetal erythroid cells. In this study, we evaluated whether limiting numbers of fetal haemopoietic progenitor cells present in an excess of maternal cells were able to overgrow the maternal component. Therefore, we used a model system in which limiting numbers of male CD34+ umbilical cord blood cells were diluted in 400 000 female CD34+ peripheral blood cells. The number of XY positive cells derived from umbilical cord blood was determined using two-colour in situ hybridization with X and Y chromosomal probes. We demonstrated a 1500-fold relative expansion of male umbilical cord blood cells over the peripheral blood component after three weeks of liquid culture, which also corresponded to the extent of expansion of CD34+ cells derived from 20-week fetal blood. However, application of the same culture protocol to maternal blood samples obtained at 7-16 weeks of gestation showed no preferential growth of fetal haemopoietic progenitor cells. This study, therefore, suggests that fetal primitive haemopoietic progenitor cells do either not circulate in maternal blood before 16 weeks of gestation, or require different combinations/concentrations of cytokines for their in vitro expansion.
Jansen
M W
MW
Department of Obstetrics and Gynecology, Academic Hospital Rotterdam Dijkzigt, Erasmus University, Rotterdam, The Netherlands. jansen@ikg.fgg.eur.nl
Korver-Hakkennes
K
K
van Leenen
D
D
Brandenburg
H
H
Wildschut
H I
HI
Wladimiroff
J W
JW
Ploemacher
R E
RE
D018952:Antigens, CD34 / Q000378:metabolism*; D002455:Cell Division; D002469:Cell Separation; D002478:Cells, Cultured; D005069:Evaluation Studies as Topic; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000502:physiology; D005315:Fetal Diseases / Q000175:diagnosis*; D005434:Flow Cytometry; D005865:Gestational Age; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D018189:Immunomagnetic Separation; D017403:In Situ Hybridization; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D011247:Pregnancy / Q000097:blood*; D011296:Prenatal Diagnosis; D015203:Reproducibility of Results; D012732:Sex Determination Analysis
D018952:Antigens, CD34