Abstract
Haematopoietic stem cells (HSCs) are a rare cell type that reconstitute the entire blood and immune systems after transplantation and can be used as a curative cell therapy for a variety of haematological diseases 1,2. However, the low number of HSCs in the body makes both biological analyses and clinical application difficult, and the limited extent to which human HSCs can be expanded ex vivo remains a substantial barrier to the wider and safer therapeutic use of HSC transplantation 3. Although various reagents have been tested in attempts to stimulate the expansion of human HSCs, cytokines have long been thought to be essential for supporting HSCs ex vivo 4. Here we report the establishment of a culture system that allows the long-term ex vivo expansion of human HSCs, achieved through the complete replacement of exogenous cytokines and albumin with chemical agonists and a caprolactam-based polymer. A phosphoinositide 3-kinase activator, in combination with a thrombopoietin-receptor agonist and the pyrimidoindole derivative UM171, were sufficient to stimulate the expansion of umbilical cord blood HSCs that are capable of serial engraftment in xenotransplantation assays. Ex vivo HSC expansion was further supported by split-clone transplantation assays and single-cell RNA-sequencing analysis. Our chemically defined expansion culture system will help to advance clinical HSC therapies.
Original language | English |
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Pages (from-to) | 127-133 |
Number of pages | 7 |
Journal | Nature |
Volume | 615 |
Issue number | 7950 |
DOIs | |
Publication status | Published - 22 Feb 2023 |
Bibliographical note
This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for detailsKeywords
- Humans
- Cell Proliferation/drug effects
- Clone Cells/cytology
- Cytokines
- Fetal Blood/cytology
- Hematopoietic Stem Cell Transplantation
- Hematopoietic Stem Cells/cytology
- Phosphatidylinositol 3-Kinases/metabolism
- Cell Culture Techniques/methods
- Albumins
- Caprolactam
- Polymers
- Receptors, Thrombopoietin
- Transplantation, Heterologous
- Single-Cell Gene Expression Analysis