Bioengineered niches that recreate physiological extracellular matrix organisation to support long-term haematopoietic stem cells

• Published in: Nature communications Vol. 15; no. 1; pp. 5791 - 18
• Main Authors: Donnelly, Hannah, Ross, Ewan, Xiao, Yinbo, Hermantara, Rio, Taqi, Aqeel F., Doherty-Boyd, W. Sebastian, Cassels, Jennifer, Tsimbouri, Penelope. M., Dunn, Karen M., Hay, Jodie, Cheng, Annie, Meek, R. M. Dominic, Jain, Nikhil, West, Christopher, Wheadon, Helen, Michie, Alison M., Peault, Bruno, West, Adam G., Salmeron-Sanchez, Manuel, Dalby, Matthew J.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 10-07-2024; Nature Publishing Group; Nature Portfolio
• Subjects: 13; 13/100; 631/80/79/750; 639/166/985; 639/638/298/923/1027; 692/308/2171; Animals; Antigens, CD34 - metabolism; Bioengineering; Bioengineering - methods; Blood; Blood diseases; Bone marrow; CD34 antigen; Cell culture; Cell Differentiation; Collagen Type I - metabolism; CRISPR; Culture media; Disorders; Extracellular matrix; Extracellular Matrix - metabolism; Hematological diseases; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - metabolism; Humanities and Social Sciences; Humans; Hydrogels - chemistry; Hypoxia-Inducible Factor 1, alpha Subunit - genetics; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism; Hypoxia-inducible factor 1a; Mechanical properties; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - metabolism; Mice; Mice, Inbred C57BL; multidisciplinary; Nestin; Nestin - genetics; Nestin - metabolism; Physiology; Polymers; Science; Science (multidisciplinary); Stem Cell Niche; Stem cell transplantation; Stem cells; Stromal cells
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-50054-0

Long-term reconstituting haematopoietic stem cells (LT-HSCs) are used to treat blood disorders via stem cell transplantation. The very low abundance of LT-HSCs and their rapid differentiation during in vitro culture hinders their clinical utility. Previous developments using stromal feeder layers, defined media cocktails, and bioengineering have enabled HSC expansion in culture, but of mostly short-term HSCs and progenitor populations at the expense of naive LT-HSCs. Here, we report the creation of a bioengineered LT-HSC maintenance niche that recreates physiological extracellular matrix organisation, using soft collagen type-I hydrogels to drive nestin expression in perivascular stromal cells (PerSCs). We demonstrate that nestin, which is expressed by HSC-supportive bone marrow stromal cells, is cytoprotective and, via regulation of metabolism, is important for HIF-1α expression in PerSCs. When CD34
+ve
HSCs were added to the bioengineered niches comprising nestin/HIF-1α expressing PerSCs, LT-HSC numbers were maintained with normal clonal and in vivo reconstitution potential, without media supplementation. We provide proof-of-concept that our bioengineered niches can support the survival of CRISPR edited HSCs. Successful editing of LT-HSCs ex vivo can have potential impact on the treatment of blood disorders.
The ability to maintain blood stem cells (HSCs) in vitro would allow us to provide better therapies for blood diseases. Here, the authors report that polymer-organised extracellular proteins, coupled to soft environments mimicking bone marrow stiffness, allow stromal cells to maintain HSCs in vitro.