Expansion of megakaryocyte progenitors from human umbilical cord blood using a new two-step separation procedure

Citation
M. Pick et al., Expansion of megakaryocyte progenitors from human umbilical cord blood using a new two-step separation procedure, BR J HAEM, 103(3), 1998, pp. 639-650
Citations number
66
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Hematology,"Cardiovascular & Hematology Research
Journal title
BRITISH JOURNAL OF HAEMATOLOGY
ISSN journal
0007-1048 → ACNP
Volume
103
Issue
3
Year of publication
1998
Pages
639 - 650
Database
ISI
SICI code
0007-1048(199812)103:3<639:EOMPFH>2.0.ZU;2-W
Abstract
Cord blood (CB) transplantation is primarily performed in children, rather than in adults, due to the low number of haemopoietic progenitor cells obta ined from the small volume of a single CB collection. Prolonged thrombocyto penia is a major problem following CB transplantation. Efforts are currentl y underway to expand the number of CB progenitor cells ex vivo, in order to enable transplantation in adults and to decrease the period of thrombocyto penia. In this study we investigated different techniques for enrichment an d expansion of megakaryocyte (Mk) progenitor cells and haemopoietic stem ce lls from CB. CBs from 20 normal deliveries were depleted of red blood cells (RBC) by dividing each sample and testing cell separation on 3% gelatin. H espan, Ficoll-Paque or a two-step 3% gelatin followed by Ficoll-Paque separ ation. The two-step procedure was found to be superior to the other methods in enrichment of the Mk progenitor cells (CFU Mk) (34.3-fold), while at th e same time retaining the number of myeloid and erythroid progenitors, CD34 (+) and CD41(+) cells. In short-term (14d) liquid culture of non-adherent n ucleated cells isolated by gelatin and Ficoll-Paque, a 40-fold expansion of clonable Mk progenitor cells was obtained in the presence of thrombopoieti n (r-hu-TPO) and stem cell factor (r-hu-SCF). In similar cultures of isolat ed CD34(+) cells, a 100-fold clonable Mk progenitor was obtained at day 14. Therefore this new technique may facilitate the ex vivo expansion of Mk pr ogenitor cells and be adopted for future use in CB transplantation.