title: Natural killer cell differentiation from hematopoietic stem cells: a comparative analysis of heparin- and stromal cell-supported methods.
abstract: Natural killer (NK) cells differentiated from hematopoietic stem cells (HSCs) may have significant clinical benefits over NK cells from adult donors, including the ability to choose alloreactive donors and potentially more robust in vivo expansion. Stromal-based methods have been used to study the differentiation of NK cells from HSCs. Stroma and cytokines support NK cell differentiation, but may face considerable regulatory hurdles. A recently reported clinical-grade, heparin-based method could serve as an alternative. How the stromal-based and heparin-based approaches compare in terms of NK cell generating efficiency or function is unknown. We show that compared with heparin-based cultures, stroma significantly increases the yield of HSC-derived NK cells by differentiating less-committed progenitors into the NK lineage. NK cells generated by both approaches were similar for most NK-activating and -inhibiting receptors. Although both approaches resulted in a phenotype consistent with CD56(bright) stage IV NK cells, heparin-based cultures favored the development of CD56(+)CD16(+) cells, whereas stroma produced more NK cell immunoglobulin-like receptor-expressing NK cells, both of which are markers of terminal maturation. At day 21, stromal-based cultures demonstrated significantly more IL-22 production, and both methods yielded similar amounts of IFN-γ production and cytotoxicity by day 35. These findings suggest that heparin-based cultures are an effective replacement for stroma and may facilitate clinical trials testing HSC-derived NK cells.
authors.affiliation: Department of Pediatrics, Division of Blood and Marrow Transplantation, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Dezell
authors.forename: Yong-Oon
authors.initials: YO
authors.lastname: Ahn
authors.forename: Jan
authors.initials: J
authors.lastname: Spanholtz
authors.forename: Hongbo
authors.initials: H
authors.lastname: Wang
authors.forename: Matthew
authors.initials: M
authors.lastname: Weeres
authors.forename: Scott
authors.initials: S
authors.lastname: Jackson
authors.forename: Sarah
authors.initials: S
authors.lastname: Cooley
authors.forename: Harry
authors.initials: H
authors.lastname: Dolstra
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Miller
authors.forename: Michael R
authors.initials: MR
authors.lastname: Verneris
mesh_terms: D018952:Antigens, CD34 / Q000096:biosynthesis / Q000276:immunology; D015415:Biomarkers / Q000032:analysis; D019002:CD56 Antigen / Q000096:biosynthesis / Q000276:immunology; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D007378:Interleukins / Q000096:biosynthesis / Q000276:immunology; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D054340:Receptors, KIR / Q000096:biosynthesis / Q000276:immunology; D017154:Stromal Cells / Q000378:metabolism*; D000097043:Interleukin-22
chemical_list: D018952:Antigens, CD34; D015415:Biomarkers; D019002:CD56 Antigen; D006493:Heparin; D007371:Interferon-gamma; D007378:Interleukins; D054340:Receptors, KIR; D000097043:Interleukin-22