In:
European Journal of Haematology, Wiley, Vol. 81, No. 3 ( 2008-09), p. 196-208
Abstract:
Aims: To study the gene profile in cord blood (CB)‐associated megakaryopoiesis. Methods: In vitro differentiation of megakaryocytes (Mks) was carried out using human CB CD34 + cells under the stimulation of recombinant human interleukin‐3, stem cell factor and thrombopoietin for 7 d, followed by thrombopoietin only for further 3 d. Lineage‐specific differentiation of Mk was examined by the expression of CD41 using flow cytometry and confocal microscopy. Total cellular RNA was extracted from day‐0 CD34 + , day‐10 CD41 + and CD41 − populations were isolated by immunomagnetic sorting respectively. Microarray was performed, and the data were analyzed using the GeneChip TM Operating System, Spotfire software and Genomatix BiblioSphere. Results: Flow cytometric analysis showed 19.44 ± 3.05% CD41 + cells at day 10 of culture. The purity of CD41 + population was enriched to 95.70 ± 4.19% after sorting. Gene expression profiling revealed an upregulation of 285 and downregulation of 53 unique genes in the CD41 + cells compared with CD41 − and CD34 + cells. Platelet‐associated genes, such as thrombospondin 1, platelet glycoprotein IIIa, etc., were highly expressed in CD41 + cells but not in CD41 − cells and CD34 + cells. Moreover, some genes that have not been reported to be associated with CB‐derived megakaryopoiesis, such as Cbl‐interacting proteins Sts‐1, protocadherin 21, etc., are found to be highly expressed in the CD41 + cells from this study. Conclusions: This study reveals a global gene expression profile of in vitro human CB‐derived megakaryopoiesis at day 10. Some of these genes may play regulatory roles during the development of CB‐derived megakaryopoeisis.
Type of Medium:
Online Resource
ISSN:
0902-4441
,
1600-0609
DOI:
10.1111/ejh.2008.81.issue-3
DOI:
10.1111/j.1600-0609.2008.01104.x
Language:
English
Publisher:
Wiley
Publication Date:
2008
detail.hit.zdb_id:
2027114-1
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