About this Biochemistry, Genetics and Molecular Biology article

Fetal Erythroblast Isolation Up to Purity from Cord Blood and Their Culture in Vitro by Giammaria Sitar; Silvia Garagna; Maurizio Zuccotti; Cristina Falcinelli; Laura Montanari; Alessandro Alfei; Giovanbattista Ippoliti; Carlo Alberto Redi; Remigio Moratti; Edoardo Ascari; Antonino Forabosco is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

Background: Erythroblasts have been the most encouraging candidate cell type for noninvasive prenatal genetic investigation. We previously showed that human erythroblasts can be recovered from bone marrow and blood bank buffy coats by a physical cell separation. In the present study, we modified our previous methodology, taking into account the peculiar behavior of erythroblasts in response to modifications of pH and osmolality of the separation medium. Methods: Twenty to forty milliters of cord blood were initially centrifuged on Ficoll/diatrizoate (1.085 g/ml). The interphase cells were further separated on a continuous density gradient (1.040-1.085 g/ml). Two different gradients were initially compared: the first was iso-osmolar and neutral, whereas the second also contained an ionic strength gradient and a pH gradient (triple gradient). A subsequent monocyte depletion was performed by using magnetic microbeads coated with anti-CD14 monoclonal antibody (mAb), and erythroblasts were purified by sedimentation velocity. Purified cells were investigated by analyses with fluorescence-activated cell sorting (FACS) and fluorescence in situ hybridization (FISH) and immunocytochemistry w

Author

Giammaria Sitar; Silvia Garagna; Maurizio Zuccotti; Cristina Falcinelli; Laura Montanari; Alessandro Alfei; Giovanbattista Ippoliti; Carlo Alberto Redi; Remigio Moratti; Edoardo Ascari; Antonino Forabosco

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; John Wiley & Sons Ltd.; Wiley (ISSN 0196-4763)

Published

1999

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)