Department of Pediatrics, Division of Blood and Marrow Transplantation, University of Minnesota, Minneapolis, Minnesota, USA.
Biol Blood Marrow Transplant. 2012 Apr;18(4):536-45. doi: 10.1016/j.bbmt.2011.11.023. Epub 2011 Dec 7.
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.
自然杀伤 (NK) 细胞由造血干细胞 (HSCs) 分化而来，与来自成人供体的 NK 细胞相比，可能具有显著的临床益处，包括选择同种反应性供体的能力和潜在更强的体内扩增能力。基于基质的方法已被用于研究 NK 细胞从 HSCs 的分化。基质和细胞因子支持 NK 细胞的分化，但可能面临相当大的监管障碍。最近报道的一种临床级别的、基于肝素的方法可能是一种替代方法。基于基质的方法和基于肝素的方法在 NK 细胞生成效率或功能方面的比较尚不清楚。我们表明，与基于肝素的培养物相比，基质通过将低分化祖细胞分化为 NK 谱系，显著增加了 HSC 衍生 NK 细胞的产量。两种方法产生的 NK 细胞在大多数 NK 激活和抑制受体方面相似。虽然两种方法都产生了与 CD56(bright) 期 IV NK 细胞一致的表型，但基于肝素的培养物有利于 CD56(+)CD16(+)细胞的发育，而基质产生了更多表达 NK 细胞免疫球蛋白样受体的 NK 细胞，这两者都是终末成熟的标志物。在第 21 天，基于基质的培养物表现出显著更多的 IL-22 产生，并且两种方法在第 35 天产生了相似数量的 IFN-γ产生和细胞毒性。这些发现表明，基于肝素的培养物是基质的有效替代品，并且可能促进测试 HSC 衍生 NK 细胞的临床试验。
