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Agitation increases expansion of cord blood hematopoietic cells and promotes their differentiation into myeloid lineage

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Abstract

Mechanical stress caused by agitation is one of the factors that can affect hematopoietic stem cell expansion in suspension bioreactors. Therefore, we have investigated the effects of agitation on umbilical cord blood hematopoietic stem cell (UCB-HSC) growth and differentiation. A comparison was made between various agitation rates (20, 40 and 60 rpm) in spinner-flask and cells cultured in glass petri dish as a static culture. Moreover, the fluid dynamic at various agitation rates of spinner-flask was analyzed to determine shear stress. The spinner-flask contained a rotational moving mixer with glass ball and was kept in tissue culture incubator. To reduce consumption of cytokines, UCB-serum was used which widely decreased the costs. Our results determined that, agitation rate at 40 rpm promoted UCB-HSCs expansion and their colony forming potential. Myeloid progenitors were the main type of cells at 40 rpm agitation rate. The results of glucose consumption and lactic acid production were in complete agreement with colony assay and expansion data and indicated the superiority of culture in spinner-flask when agitated at 40 rpm over to other agitation speeds and also static culture. Cell viability and colony count was affected by changing the agitation speed. We assume that changes in cell growth resulted from the effect of shear stress directly on cell viability, and indirectly on signaling pathways that influence the cells to differentiate.

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Acknowledgments

The authors thank Mr. Fazel S. Sahraneshin for his excellent technical assistance and helpful advices at flow cytometry data. This study was funded by a grant provided from Royan Institute.

Conflict of interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Department of Chemical and Petroleum Engineering, Sharif University of Technology, Tehran, 11155-9465, Iran

    Hasti Hosseinizand & Mohammad J. Abdekhodaie

  2. Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

    Hasti Hosseinizand & Marzieh Ebrahimi

Authors
  1. Hasti Hosseinizand
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  2. Marzieh Ebrahimi
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  3. Mohammad J. Abdekhodaie
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Correspondence to Marzieh Ebrahimi or Mohammad J. Abdekhodaie.

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Hosseinizand, H., Ebrahimi, M. & Abdekhodaie, M.J. Agitation increases expansion of cord blood hematopoietic cells and promotes their differentiation into myeloid lineage. Cytotechnology 68, 969–978 (2016). https://doi.org/10.1007/s10616-015-9851-3

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  • DOI: https://doi.org/10.1007/s10616-015-9851-3

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