Bioactive poly(ethylene glycol) hydrogels to recapitulate the HSC niche and facilitate HSC expansion in culture.
Hematopoietic stem cells (HSCs) have been used therapeutically for decades, yet their widespread clinical use is hampered by the inability to expand HSCs successfully in vitro. In culture, HSCs rapidly differentiate and lose their ability to self-renew. We hypothesize that by mimicking aspects of the bone marrow microenvironment in vitro we can better control the expansion and differentiation of these cells. In this work, derivatives of poly(ethylene glycol) diacrylate hydrogels were used as a culture substrate for hematopoietic stem and progenitor cell (HSPC) populations. Key HSC cytokines, stem cell factor (SCF) and interferon-γ (IFNγ), as well as the cell adhesion ligands RGDS and connecting segment 1 were covalently immobilized onto the surface of the hydrogels. With the use of SCF and IFNγ, we observed significant expansion of HSPCs, ∼97 and ∼104 fold respectively, while maintaining c-kit(+) lin(-) and c-kit(+) Sca1(+) lin(-) (KSL) populations and the ability to form multilineage colonies after 14 days. HSPCs were also encapsulated within degradable poly(ethylene glycol) hydrogels for three-dimensional culture. After expansion in hydrogels, ∼60% of cells were c-kit(+), demonstrating no loss in the proportion of these cells over the 14 day culture period, and ∼50% of colonies formed were multilineage, indicating that the cells retained their differentiation potential. The ability to tailor and use this system to support HSC growth could have implications on the future use of HSCs and other blood cell types in a clinical setting.
Department of Bioengineering, Rice University, Houston, Texas.
Maude L
ML
Cuchiara
Department of Internal Medicine, Yale Cardiovascular Research Center, Vascular Biology and Therapeutics Program and Yale Stem Cell Center, Yale University School of Medicine, New Haven, Connecticut.|Departments of Pediatrics and Molecular and Cellular Biology, Children's Nutrition Research Center and Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas.
Süleyman
S
Coşkun
Department of Bioengineering, Rice University, Houston, Texas.
Omar A
OA
Banda
Department of Bioengineering, Rice University, Houston, Texas.
Kelsey L
KL
Horter
Department of Internal Medicine, Yale Cardiovascular Research Center, Vascular Biology and Therapeutics Program and Yale Stem Cell Center, Yale University School of Medicine, New Haven, Connecticut.|Departments of Pediatrics and Molecular and Cellular Biology, Children's Nutrition Research Center and Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas.
Karen K
KK
Hirschi
Department of Bioengineering, Rice University, Houston, Texas. jennifer.l.west@duke.edu.|Department of Biomedical Engineering, Duke University, Room 1427, FCIEMAS, 101 Science Dr., Box 90281, Durham, North Carolina, 27708. jennifer.l.west@duke.edu.
Jennifer L
JL
West
D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D020100:Hydrogels; D007371:Interferon-gamma / Q000378:metabolism; D011092:Polyethylene Glycols; D019089:Stem Cell Factor / Q000378:metabolism
D003470:Culture Media; D020100:Hydrogels; D019089:Stem Cell Factor; D011092:Polyethylene Glycols; D007371:Interferon-gamma
expansion; hematopoietic stem cell; hydrogel; interferon gamma; poly(ethylene glycol); stem cell factor