Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells (hESCs) and and Induced Pluripotent Stem Cells (iPSCs)
Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells (hESCs) and and Induced Pluripotent Stem Cells (iPSCs)
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DOI：
10.3791/50337
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发表时间：
2013-04-01
影响因子：
1.2
通讯作者：
Kaufman, Dan S.
中科院分区：
文献类型：
--
作者：
Bock, Allison M.;Knorr, David;Kaufman, Dan S.
我们提出了一种使用无饲养层方法从未分化的hESC和iPSC中获得自然杀伤（NK）细胞的方法。这种方法在培养4周后产生高水平的NK细胞，并且可以用人工抗原呈递细胞进行进一步的2-log扩增。在该系统中开发的hESC和iPSC衍生的NK细胞具有成熟的表型和功能。大量可遗传修饰的NK细胞的产生适用于基本机制以及抗肿瘤研究。萤火虫荧光素酶在hESC衍生的NK细胞中的表达允许非侵入性方法来跟踪NK细胞植入、分布和功能。我们还描述了一种双重成像方案，允许单独监测两个不同的细胞群，以更清楚地表征它们在体内的相互作用。这种衍生、扩增和双重体内成像的方法为产生NK细胞及其评价提供了可靠的方法，这对于改善目前的NK细胞过继疗法是必要的。
We present a method for deriving natural killer (NK) cells from undifferentiated hESCs and iPSCs using a feeder-free approach. This method gives rise to high levels of NK cells after 4 weeks culture and can undergo further 2-log expansion with artificial antigen presenting cells. hESC-and iPSC-derived NK cells developed in this system have a mature phenotype and function. The production of large numbers of genetically modifiable NK cells is applicable for both basic mechanistic as well as anti-tumor studies. Expression of firefly luciferase in hESC-derived NK cells allows a non-invasive approach to follow NK cell engraftment, distribution, and function. We also describe a dual-imaging scheme that allows separate monitoring of two different cell populations to more distinctly characterize their interactions in vivo. This method of derivation, expansion, and dual in vivo imaging provides a reliable approach for producing NK cells and their evaluation which is necessary to improve current NK cell adoptive therapies.