Physioxia improves the selectivity of hematopoietic stem cell expansion cultures

Kyomi J Igarashi; Iwo Kucinski; Yan Yi Chan; Tze-Kai Tan; Hwei Minn Khoo; David Kealy; Joydeep Bhadury; Ian Hsu; Pui Yan Ho; Kouta Niizuma; John W Hickey; Garry Nolan; Katherine S Bridge; Agnieszka Czechowicz; Berthold Gottgens; Hiromitsu Nakauchi; Adam C Wilkinson

doi:10.1182/bloodadvances.2023009668

Physioxia improves the selectivity of hematopoietic stem cell expansion cultures

Kyomi J Igarashi, Iwo Kucinski, Yan Yi Chan, Tze-Kai Tan, Hwei Minn Khoo, David Kealy, Joydeep Bhadury, Ian Hsu, Pui Yan Ho, Kouta Niizuma, John W Hickey, Garry Nolan, Katherine S Bridge, Agnieszka Czechowicz, Berthold Gottgens, Hiromitsu Nakauchi, Adam C Wilkinson^*

^*Corresponding author for this work

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Hematopoietic stem cells (HSCs) are a rare hematopoietic cell type that can entirely reconstitute the blood and immune systems following transplantation. Allogeneic HSC transplantation (HSCT) is used clinically as a curative therapy for a range of hematolymphoid diseases, but remains a high-risk therapy due to potential side effects including poor graft function and graft-vs-host disease (GvHD). Ex vivo HSC expansion has been suggested as an approach to improve hematopoietic reconstitution from low-cell dose grafts. Here, we demonstrate that we can improve the selectivity of polyvinyl alcohol (PVA)-based mouse HSC cultures through the use of physioxic culture conditions. Single-cell transcriptomic analysis confirmed inhibition of lineage-committed progenitor cells in physioxic cultures. Long-term physioxic expansion also afforded culture-based ex vivo HSC selection from whole bone marrow, spleen, and embryonic tissues. Furthermore, we provide evidence that HSC-selective ex vivo cultures deplete GvHD-causing T cells and that this approach can be combined with genotoxic-free antibody-based conditioning HSCT approaches. Our results offer a simple approach to improve PVA-based HSC cultures and the underlying molecular phenotype, as well as highlight the potential translational implications of selective HSC expansion systems for allogeneic HSCT.

Original language	English
Journal	Blood Advances
Early online date	21 Feb 2023
DOIs	https://doi.org/10.1182/bloodadvances.2023009668
Publication status	E-pub ahead of print - 21 Feb 2023

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10.1182/bloodadvances.2023009668

Physioxia improves the selectivity of hematopoietic stem cell expansion cultures
© 2023 American Society of Hematology. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details
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Cite this

Igarashi, K. J., Kucinski, I., Chan, Y. Y., Tan, T.-K., Khoo, H. M., Kealy, D., Bhadury, J., Hsu, I., Ho, P. Y., Niizuma, K., Hickey, J. W., Nolan, G., Bridge, K. S., Czechowicz, A., Gottgens, B., Nakauchi, H., & Wilkinson, A. C. (2023). Physioxia improves the selectivity of hematopoietic stem cell expansion cultures. Blood Advances. Advance online publication. https://doi.org/10.1182/bloodadvances.2023009668

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abstract = "Hematopoietic stem cells (HSCs) are a rare hematopoietic cell type that can entirely reconstitute the blood and immune systems following transplantation. Allogeneic HSC transplantation (HSCT) is used clinically as a curative therapy for a range of hematolymphoid diseases, but remains a high-risk therapy due to potential side effects including poor graft function and graft-vs-host disease (GvHD). Ex vivo HSC expansion has been suggested as an approach to improve hematopoietic reconstitution from low-cell dose grafts. Here, we demonstrate that we can improve the selectivity of polyvinyl alcohol (PVA)-based mouse HSC cultures through the use of physioxic culture conditions. Single-cell transcriptomic analysis confirmed inhibition of lineage-committed progenitor cells in physioxic cultures. Long-term physioxic expansion also afforded culture-based ex vivo HSC selection from whole bone marrow, spleen, and embryonic tissues. Furthermore, we provide evidence that HSC-selective ex vivo cultures deplete GvHD-causing T cells and that this approach can be combined with genotoxic-free antibody-based conditioning HSCT approaches. Our results offer a simple approach to improve PVA-based HSC cultures and the underlying molecular phenotype, as well as highlight the potential translational implications of selective HSC expansion systems for allogeneic HSCT.",

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AU - Igarashi, Kyomi J

AU - Kucinski, Iwo

AU - Chan, Yan Yi

AU - Tan, Tze-Kai

AU - Khoo, Hwei Minn

AU - Kealy, David

AU - Bhadury, Joydeep

AU - Hsu, Ian

AU - Ho, Pui Yan

AU - Niizuma, Kouta

AU - Hickey, John W

AU - Nolan, Garry

AU - Bridge, Katherine S

AU - Czechowicz, Agnieszka

AU - Gottgens, Berthold

AU - Nakauchi, Hiromitsu

AU - Wilkinson, Adam C

PY - 2023/2/21

Y1 - 2023/2/21

N2 - Hematopoietic stem cells (HSCs) are a rare hematopoietic cell type that can entirely reconstitute the blood and immune systems following transplantation. Allogeneic HSC transplantation (HSCT) is used clinically as a curative therapy for a range of hematolymphoid diseases, but remains a high-risk therapy due to potential side effects including poor graft function and graft-vs-host disease (GvHD). Ex vivo HSC expansion has been suggested as an approach to improve hematopoietic reconstitution from low-cell dose grafts. Here, we demonstrate that we can improve the selectivity of polyvinyl alcohol (PVA)-based mouse HSC cultures through the use of physioxic culture conditions. Single-cell transcriptomic analysis confirmed inhibition of lineage-committed progenitor cells in physioxic cultures. Long-term physioxic expansion also afforded culture-based ex vivo HSC selection from whole bone marrow, spleen, and embryonic tissues. Furthermore, we provide evidence that HSC-selective ex vivo cultures deplete GvHD-causing T cells and that this approach can be combined with genotoxic-free antibody-based conditioning HSCT approaches. Our results offer a simple approach to improve PVA-based HSC cultures and the underlying molecular phenotype, as well as highlight the potential translational implications of selective HSC expansion systems for allogeneic HSCT.

AB - Hematopoietic stem cells (HSCs) are a rare hematopoietic cell type that can entirely reconstitute the blood and immune systems following transplantation. Allogeneic HSC transplantation (HSCT) is used clinically as a curative therapy for a range of hematolymphoid diseases, but remains a high-risk therapy due to potential side effects including poor graft function and graft-vs-host disease (GvHD). Ex vivo HSC expansion has been suggested as an approach to improve hematopoietic reconstitution from low-cell dose grafts. Here, we demonstrate that we can improve the selectivity of polyvinyl alcohol (PVA)-based mouse HSC cultures through the use of physioxic culture conditions. Single-cell transcriptomic analysis confirmed inhibition of lineage-committed progenitor cells in physioxic cultures. Long-term physioxic expansion also afforded culture-based ex vivo HSC selection from whole bone marrow, spleen, and embryonic tissues. Furthermore, we provide evidence that HSC-selective ex vivo cultures deplete GvHD-causing T cells and that this approach can be combined with genotoxic-free antibody-based conditioning HSCT approaches. Our results offer a simple approach to improve PVA-based HSC cultures and the underlying molecular phenotype, as well as highlight the potential translational implications of selective HSC expansion systems for allogeneic HSCT.

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