Cellular and Molecular Life Sciences

, Volume 67, Issue 10, pp 1713–1722 | Cite as

Role of Chk1 in the differentiation program of hematopoietic stem cells

  • Laura CarrassaEmail author
  • Elisa Montelatici
  • Lorenza Lazzari
  • Stefano Zangrossi
  • Matteo Simone
  • Massimo Broggini
  • Giovanna Damia
Research Article


Hematopoietic stem cells (HSC) isolated from umbilical cord blood (UCB) were treated with ionizing radiation (IR) and sensitivity and IR induced checkpoints activation were investigated. No difference in the sensitivity and in the activation of DNA damage pathways was observed between CD133+ HSC and cells derived from them after ex vivo expansion. Chk1 protein was very low in freshly isolated CD133+ cells, and undetectable in ex vivo expanded UCB CD133+ cells. Chk1 was expressed only on day 3 of the ex vivo expansion. This pattern of Chk1 expression was corroborated in CD133+ cells isolated from peripheral blood apheresis collected from an healthy donor. Treatment with a specific Chk1 inhibitor resulted in a strong reduction in the percentage of myeloid precursors (CD33+) and an increase in the percentage of lymphoid precursors (CD38+) compared to untreated cells, suggesting a possible role for Chk1 in the differentiation program of UCB CD133+ HSC.


Stem and progenitor cells Umbilical cord blood Chk1 DNA damage Differentiation 



We thank Dr Patrick Casara and Dr John Hickman who respectively synthesized and kindly provided the Chk1 inhibitor compound. A special thank to JD Baggott that kindly edited the paper. This study was supported by grants from “Fondazione I1 Sangue”, Ministero della Salute, Istituto Superiore di Sanità, Sixth FP “Thercord”, Foundation Novussanguis and Jérome Lejeune. The generous contribution of the Italian Association for Cancer Research and of FIRB is also gratefully acknowledged.

Supplementary material

18_2010_274_MOESM1_ESM.tif (174 kb)
UCB CD133+ cell growth curve from isolation up to day 14 of ex vivo expansion with the cytokine cocktail. Mean ± SD of three experiments performed from UCB of three different donors. The bottom panel in the figure gives a larger view of the growth curve from day 0 to day 6 of ex vivo expansion (TIFF 174 kb)
18_2010_274_MOESM2_ESM.tif (146 kb)
Cytotoxic effect of IR in UCB CD133+ cells the day of isolation (♦), in the ex vivo expanded population at day 3 from isolation (■) and in the partly differentiated cells at day 14 from isolation (●). Data are expressed as percentages of the untreated control sample; mean ± SD of 3 different experiments done in triplicate (TIFF 146 kb)
18_2010_274_MOESM3_ESM.tif (567 kb)
Western blot analysis showing Chk1, pS317Chk1, pCdc25C, Cdc25C and Ran protein levels of UCB CD133+ cells, untreated or treated with the Chk1 inhibitor as described in Materials and Methods (TIFF 566 kb)
18_2010_274_MOESM4_ESM.tif (313 kb)
(A) Upper panel: Table representing the relative proportion of cells positive for the differentiation markers selected both in untreated and in Chk1 inhibitor AZD- 7762 treated cells at day 14 of the ex vivo expansion. Lower panel: Western Blot Analysis showing pS317 Chk1 and total Chk1 protein levels of UCB CD 133+ cells untreated or treated with AZD 7762. (B) Upper panel: Table representing the decrease in percentage of CD 33 positive HL 60 cells after treatment with CHIR-124 (120 nM) and AZD 7762 (500 nM). Lower panel: Western Blot Analysis showing pS317 Chk1 and total Chk1 protein levels of HL-60 cells untreated or treated with AZD 7762 and CHIR-124 (TIFF 312 kb)


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Copyright information

© Springer Basel AG 2010

Authors and Affiliations

  • Laura Carrassa
    • 1
    Email author
  • Elisa Montelatici
    • 2
  • Lorenza Lazzari
    • 2
  • Stefano Zangrossi
    • 2
  • Matteo Simone
    • 1
  • Massimo Broggini
    • 1
  • Giovanna Damia
    • 1
  1. 1.Laboratory of Molecular Pharmacology, Department of OncologyIstituto di Ricerche Farmacologiche “Mario Negri”MilanItaly
  2. 2.Cell Factory, Centre for Transfusion Medicine, Cell Therapy and Cryobiology, Department of Regenerative MedicineOspedale Maggiore PoliclinicoMilanItaly

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