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Reconstruction of bone marrow microenvironment for expansion of hematopoietic stem cells by a histone deacetylase inhibitor

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Abstract

Ex vivo expansion of hematopoietic stem cells (HSCs) is an approach for overcoming cell insufficiency for umbilical cord blood transplantation. It was suggested that in common ex vivo cultures, the stemness specificity of HSCs is rapidly reducing due to DNA hypermethylation. Here, Nicotinamide (NAM), a DNA methyltransferase and histone deacetylase inhibitor, is used with a bioengineered Bone Marrow-like niche (BLN) for HSC ex vivo expansion. The CFSE cell proliferation assay was used for tracking HSCs division. qRT-PCR was conducted to assay the HOXB4 mRNA expression levels. The morphology of BLN-cultured cells was analyzed using scanning electron microscopy (SEM). NAM boosted the induction of HSC proliferation in the BLN group compared to the control group. In addition, the ability of HSCs to colonize was more significant in the BLN group than in the control group. Our data suggest that the presence of NAM in bioengineered niches promotes HSC proliferation. The presented approach showed that small molecules could be used in the clinical setting to overcome the limited number of CD34+ cells in cord blood units.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Abbreviations

BM:

Bone marrow

BLN:

BM-like niche

BFU-E:

Burst-Forming Unit-Erythroid

CFU-GEMM:

Colony-Forming Unit—Granulocyte, Erythroid, Macrophage, Megakaryocyte

CFU-GM:

Colony-Forming Unit—Granulocyte–Macrophage

CFSE:

CarboxyFluorescein diacetate N- Succinilamidyle Ester

DBM:

Demineralized Bone Matrix

HSC:

Hematopoietic Stem Cell

HDI:

Histone Deacetylase Inhibitors

MSC:

Mesenchymal Stem cell

NAM:

Nicotinamide

NAM-CY:

Nicotinamide, and cytokines

SEM:

Scanning Electron Microscopy

TCC:

Traditional culture condition

2D:

2-Dimensional

3D:

3-Dimensional

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Funding

This study was supported by High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.

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Authors and Affiliations

  1. Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, IBTO Building, Hemmat Expressway, Tehran, Iran

    Maryam Haj Ali Askari, Majid Shahabi & Mahin Nikougoftar Zarif

  2. Department of Hematology and Blood Bank, Iran University of Medical Sciences, Tehran, Iran

    Amir Asri Kojabad

  3. Department of Medicine, Center for Hematology and Regenerative Medicin, 14183, Stockholm, Sweden

    Mahin Nikougoftar Zarif

  4. Cell Therapy Department, XNKtheraeutics Company, Stockholm, Sweden

    Mahin Nikougoftar Zarif

Authors
  1. Maryam Haj Ali Askari
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  2. Majid Shahabi
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  3. Amir Asri Kojabad
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  4. Mahin Nikougoftar Zarif
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Contributions

MHAA, MS and MNZ wrote the main manuscript text and AAK prepared figures. All authors reviewed the manuscript.

Corresponding author

Correspondence to Mahin Nikougoftar Zarif.

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Askari, M.H.A., Shahabi, M., Kojabad, A.A. et al. Reconstruction of bone marrow microenvironment for expansion of hematopoietic stem cells by a histone deacetylase inhibitor. Cytotechnology 75, 195–206 (2023). https://doi.org/10.1007/s10616-022-00564-w

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