Generation of transgene-free hematopoietic stem cells from human induced pluripotent stem cells

Piau, Olivier; Brunet-Manquat, Mathias; L’Homme, Bruno; Petit, Laurence; Birebent, Brigitte; Linard, Christine; Moeckes, Laetitia; Zuliani, Thomas; Lapillonne, Hélène; Benderitter, Marc; Douay, Luc; Chapel, Alain; Guyonneau-Harmand, Laurence; Jaffredo, Thierry

doi:10.1016/j.stem.2023.11.002

Cited by 10 publications

(1 citation statement)

References 64 publications

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“…hPSCs, such as hESCs, are ideal seed cells for obtaining large numbers of HSPCs and for scalable manufacturing of human hematopoietic or BM organoids. Accumulating evidence demonstrates that hESCs can differentiate into HSPCs and multilineage blood cells [ 36 , 37 ]. Therefore, we used a stepwise induction protocol to generate human CD34 + HSPCs from hESCs.…”

Section: Discussionmentioning

confidence: 99%

Establishment of human hematopoietic organoids for evaluation of hematopoietic injury and regeneration effect

Chen,

Li,

et al. 2024

Stem Cell Res Ther

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Background Human hematopoietic organoids have a wide application value for modeling human bone marrow diseases, such as acute hematopoietic radiation injury. However, the manufacturing of human hematopoietic organoids is an unaddressed challenge because of the complexity of hematopoietic tissues. Methods To manufacture hematopoietic organoids, we obtained CD34+ hematopoietic stem and progenitor cells (HSPCs) from human embryonic stem cells (hESCs) using stepwise induction and immunomagnetic bead-sorting. We then mixed these CD34+ HSPCs with niche-related cells in Gelatin-methacryloyl (GelMA) to form a three-dimensional (3D) hematopoietic organoid. Additionally, we investigated the effects of radiation damage and response to granulocyte colony-stimulating factor (G-CSF) in hematopoietic organoids. Results The GelMA hydrogel maintained the undifferentiated state of hESCs-derived HSPCs by reducing intracellular reactive oxygen species (ROS) levels. The established hematopoietic organoids in GelMA with niche-related cells were composed of HSPCs and multilineage blood cells and demonstrated the adherence of hematopoietic cells to niche cells. Notably, these hematopoietic organoids exhibited radiation-induced hematopoietic cell injury effect, including increased intracellular ROS levels, γ-H2AX positive cell percentages, and hematopoietic cell apoptosis percentages. Moreover, G-CSF supplementation in the culture medium significantly improved the survival of HSPCs and enhanced myeloid cell regeneration in these hematopoietic organoids after radiation. Conclusions These findings substantiate the successful manufacture of a preliminary 3D hematopoietic organoid from hESCs-derived HSPCs, which was utilized for modeling hematopoietic radiation injury and assessing the radiation-mitigating effects of G-CSF in vitro. Our study provides opportunities to further aid in the standard and scalable production of hematopoietic organoids for disease modeling and drug testing.

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Section: Discussionmentioning

confidence: 99%