Selective modulation of the cyclin B/CDK1 and cyclin D/CDK4 complexes during in vitro human megakaryocyte development

Alessandra Bassini, Sabina Pierpaoli, Elisabetta Falcieri, Marco Vitale, Lia Guidotti, Silvano Capitani, Giorgio Zauli

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Mammalian megakaryocyte development is characterized by a progressive accumulation of cells exhibiting a polylobated nucleus with a polyploid DNA content. In this study human megakaryocytes were obtained from CD34+ haemopoietic progenitors by in vitro liquid culture in the presence of 100 ng/ml of recombinant thrombopoietin (TPO). Ultrastructural examination of polyploid megakaryocytes showed the presence of a large number of centrioles, the breakdown of the nuclear envelope, and the progressive chromatin condensation, all aspects characteristic of mitosis. At both indirect immunofluorescence and Western blot analyses, cyclin B and its related cyclin-dependent kinase (CDK)1, which forms the mitosis promoting factor (MPF), showed an increased expression in maturating megakaryoblasts and megakaryocytes (day 8 of culture) with respect to freshly isolated CD34+ progenitors. This expression tended to decline in fully developed megakaryocytes (day 15 of culture). The amount of cyclin D and of the related CDK4, governing the G1 phase of the cell cycle, increased during megakaryocyte development, maintaining high levels of expression also in mature megakaryocytes. These results indicate that megakaryocyte polyploidization depends on a true, although incomplete, mitotic process, and that cyclin D/CDK4 probably plays a crucial role throughout megakaryocytopoiesis.

Original languageEnglish
Pages (from-to)820-828
Number of pages9
JournalBritish Journal of Haematology
Volume104
Issue number4
Publication statusPublished - 1999

Fingerprint

Cyclin D
Cyclin B
Megakaryocytes
Human Development
Polyploidy
Maturation-Promoting Factor
Megakaryocyte Progenitor Cells
CDC2 Protein Kinase
Thrombopoiesis
Centrioles
Thrombopoietin
In Vitro Techniques
Nuclear Envelope
G1 Phase
Indirect Fluorescent Antibody Technique
Mitosis
Chromatin
Cell Cycle
Western Blotting
DNA

Keywords

  • CDK
  • Cyclins
  • Megakaryocytes
  • TPO

ASJC Scopus subject areas

  • Hematology

Cite this

Bassini, A., Pierpaoli, S., Falcieri, E., Vitale, M., Guidotti, L., Capitani, S., & Zauli, G. (1999). Selective modulation of the cyclin B/CDK1 and cyclin D/CDK4 complexes during in vitro human megakaryocyte development. British Journal of Haematology, 104(4), 820-828.

Selective modulation of the cyclin B/CDK1 and cyclin D/CDK4 complexes during in vitro human megakaryocyte development. / Bassini, Alessandra; Pierpaoli, Sabina; Falcieri, Elisabetta; Vitale, Marco; Guidotti, Lia; Capitani, Silvano; Zauli, Giorgio.

In: British Journal of Haematology, Vol. 104, No. 4, 1999, p. 820-828.

Research output: Contribution to journalArticle

Bassini, A, Pierpaoli, S, Falcieri, E, Vitale, M, Guidotti, L, Capitani, S & Zauli, G 1999, 'Selective modulation of the cyclin B/CDK1 and cyclin D/CDK4 complexes during in vitro human megakaryocyte development', British Journal of Haematology, vol. 104, no. 4, pp. 820-828.
Bassini A, Pierpaoli S, Falcieri E, Vitale M, Guidotti L, Capitani S et al. Selective modulation of the cyclin B/CDK1 and cyclin D/CDK4 complexes during in vitro human megakaryocyte development. British Journal of Haematology. 1999;104(4):820-828.
Bassini, Alessandra ; Pierpaoli, Sabina ; Falcieri, Elisabetta ; Vitale, Marco ; Guidotti, Lia ; Capitani, Silvano ; Zauli, Giorgio. / Selective modulation of the cyclin B/CDK1 and cyclin D/CDK4 complexes during in vitro human megakaryocyte development. In: British Journal of Haematology. 1999 ; Vol. 104, No. 4. pp. 820-828.
@article{16c6ffa2e39a4adabd54b971420fa210,
title = "Selective modulation of the cyclin B/CDK1 and cyclin D/CDK4 complexes during in vitro human megakaryocyte development",
abstract = "Mammalian megakaryocyte development is characterized by a progressive accumulation of cells exhibiting a polylobated nucleus with a polyploid DNA content. In this study human megakaryocytes were obtained from CD34+ haemopoietic progenitors by in vitro liquid culture in the presence of 100 ng/ml of recombinant thrombopoietin (TPO). Ultrastructural examination of polyploid megakaryocytes showed the presence of a large number of centrioles, the breakdown of the nuclear envelope, and the progressive chromatin condensation, all aspects characteristic of mitosis. At both indirect immunofluorescence and Western blot analyses, cyclin B and its related cyclin-dependent kinase (CDK)1, which forms the mitosis promoting factor (MPF), showed an increased expression in maturating megakaryoblasts and megakaryocytes (day 8 of culture) with respect to freshly isolated CD34+ progenitors. This expression tended to decline in fully developed megakaryocytes (day 15 of culture). The amount of cyclin D and of the related CDK4, governing the G1 phase of the cell cycle, increased during megakaryocyte development, maintaining high levels of expression also in mature megakaryocytes. These results indicate that megakaryocyte polyploidization depends on a true, although incomplete, mitotic process, and that cyclin D/CDK4 probably plays a crucial role throughout megakaryocytopoiesis.",
keywords = "CDK, Cyclins, Megakaryocytes, TPO",
author = "Alessandra Bassini and Sabina Pierpaoli and Elisabetta Falcieri and Marco Vitale and Lia Guidotti and Silvano Capitani and Giorgio Zauli",
year = "1999",
language = "English",
volume = "104",
pages = "820--828",
journal = "British Journal of Haematology",
issn = "0007-1048",
publisher = "John Wiley & Sons, Ltd (10.1111)",
number = "4",

}

TY - JOUR

T1 - Selective modulation of the cyclin B/CDK1 and cyclin D/CDK4 complexes during in vitro human megakaryocyte development

AU - Bassini, Alessandra

AU - Pierpaoli, Sabina

AU - Falcieri, Elisabetta

AU - Vitale, Marco

AU - Guidotti, Lia

AU - Capitani, Silvano

AU - Zauli, Giorgio

PY - 1999

Y1 - 1999

N2 - Mammalian megakaryocyte development is characterized by a progressive accumulation of cells exhibiting a polylobated nucleus with a polyploid DNA content. In this study human megakaryocytes were obtained from CD34+ haemopoietic progenitors by in vitro liquid culture in the presence of 100 ng/ml of recombinant thrombopoietin (TPO). Ultrastructural examination of polyploid megakaryocytes showed the presence of a large number of centrioles, the breakdown of the nuclear envelope, and the progressive chromatin condensation, all aspects characteristic of mitosis. At both indirect immunofluorescence and Western blot analyses, cyclin B and its related cyclin-dependent kinase (CDK)1, which forms the mitosis promoting factor (MPF), showed an increased expression in maturating megakaryoblasts and megakaryocytes (day 8 of culture) with respect to freshly isolated CD34+ progenitors. This expression tended to decline in fully developed megakaryocytes (day 15 of culture). The amount of cyclin D and of the related CDK4, governing the G1 phase of the cell cycle, increased during megakaryocyte development, maintaining high levels of expression also in mature megakaryocytes. These results indicate that megakaryocyte polyploidization depends on a true, although incomplete, mitotic process, and that cyclin D/CDK4 probably plays a crucial role throughout megakaryocytopoiesis.

AB - Mammalian megakaryocyte development is characterized by a progressive accumulation of cells exhibiting a polylobated nucleus with a polyploid DNA content. In this study human megakaryocytes were obtained from CD34+ haemopoietic progenitors by in vitro liquid culture in the presence of 100 ng/ml of recombinant thrombopoietin (TPO). Ultrastructural examination of polyploid megakaryocytes showed the presence of a large number of centrioles, the breakdown of the nuclear envelope, and the progressive chromatin condensation, all aspects characteristic of mitosis. At both indirect immunofluorescence and Western blot analyses, cyclin B and its related cyclin-dependent kinase (CDK)1, which forms the mitosis promoting factor (MPF), showed an increased expression in maturating megakaryoblasts and megakaryocytes (day 8 of culture) with respect to freshly isolated CD34+ progenitors. This expression tended to decline in fully developed megakaryocytes (day 15 of culture). The amount of cyclin D and of the related CDK4, governing the G1 phase of the cell cycle, increased during megakaryocyte development, maintaining high levels of expression also in mature megakaryocytes. These results indicate that megakaryocyte polyploidization depends on a true, although incomplete, mitotic process, and that cyclin D/CDK4 probably plays a crucial role throughout megakaryocytopoiesis.

KW - CDK

KW - Cyclins

KW - Megakaryocytes

KW - TPO

UR - http://www.scopus.com/inward/record.url?scp=0033029073&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033029073&partnerID=8YFLogxK

M3 - Article

C2 - 10192445

AN - SCOPUS:0033029073

VL - 104

SP - 820

EP - 828

JO - British Journal of Haematology

JF - British Journal of Haematology

SN - 0007-1048

IS - 4

ER -