Domogala Anna, Madrigal J Alejandro, Saudemont Aurore
Anthony Nolan Research Institute, London, United Kingdom; University College London, London, United Kingdom.
Anthony Nolan Research Institute, London, United Kingdom; University College London, London, United Kingdom.
Cytotherapy. 2016 Jun;18(6):754-9. doi: 10.1016/j.jcyt.2016.02.008. Epub 2016 Apr 15.
Natural killer (NK) cells offer the potential for a powerful cellular immunotherapy because they can target malignant cells without being direct effectors of graft-versus-host disease. We have previously shown that high numbers of functional NK cells can be differentiated in vitro from umbilical cord blood (CB) CD34(+) cells. To develop a readily available, off-the-shelf cellular product, it is essential that NK cells differentiated in vitro can be frozen and thawed while maintaining the same phenotype and functions.
We evaluated the phenotype and function of fresh and frozen NK cells differentiated in vitro. We also assessed whether the concentration of NK cells at the time of freezing had an impact on cell viability.
We found that cell concentration of NK cells at the time of freezing did not have an impact on their viability and on cell recovery post-thaw. Moreover, freezing of differentiated NK cells in vitro did not affect their phenotype, cytotoxicity and degranulation capacity toward K562 cells, cytokine production and proliferation.
We are therefore able to generate large numbers of functional NK cells from CB CD34(+) cells that maintain the same phenotype and function post-cryopreservation, which will allow for multiple infusions of a highly cytotoxic NK cell product.
自然杀伤（NK）细胞具有强大的细胞免疫治疗潜力，因为它们能够靶向恶性细胞，而不会引发移植物抗宿主病。我们之前已经证明，大量功能性NK细胞可在体外从脐带血（CB）CD34(+)细胞分化而来。为开发一种随时可用的现成细胞产品，体外分化的NK细胞在冷冻和解冻后仍保持相同的表型和功能至关重要。
我们评估了体外分化的新鲜和冷冻NK细胞的表型和功能。我们还评估了冷冻时NK细胞的浓度是否对细胞活力有影响。
我们发现冷冻时NK细胞的细胞浓度对其活力和解冻后的细胞回收率没有影响。此外，体外分化的NK细胞冷冻不会影响其表型、对K562细胞的细胞毒性和脱颗粒能力、细胞因子产生及增殖。
因此，我们能够从CB CD34(+)细胞中生成大量功能性NK细胞，这些细胞在冷冻保存后保持相同的表型和功能，这将允许多次输注高细胞毒性的NK细胞产品。
