Peighambarzadeh Farzaneh, Najafalizadeh Anahita, Esmaeil Nafiseh, Rezaei Abbas, Ashrafi Farzaneh, Ganjalikhani Hakemi Mazdak
Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Hematology Division, Department of Internal Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Avicenna J Med Biotechnol. 2020 Jan-Mar;12(1):17-23.
Regarding to the increase of cancer deaths in recent years and disability of common therapies to eradicate cancers, as well as expansion of Natural Killer (NK) cell therapy, it seems so vital to find new useful therapies against cancers. Breast cancer is the second main cause of cancer death among women. As it is impossible for a majority of patients to receive NK cell therapy, an attempt was made to establish a low-cost and efficient method for expanding and activating NK cells against breast cancer cell line (MCF7).
NK cells were isolated from Peripheral Blood Mononuclear Cells (PBMCs) applying either MACS based NK cell enrichment kit or antibodies and complement as cytotoxic method. Then, the NK cells were cultured in Stem Cell Growth Medium (SCGM) with feeder layer (irradiated PBMCs) along with PHA or OKT3. IL-2, IL-15 and IL-21 were used to expand NK cells and finally their cytotoxic activity was investigated by flow cytometry.
Highly pure NK cells were obtained and no significant difference between the two isolation methods was found. Using IL-2 plus IL-15, the number of NK cells increased up to100 fold after 16 days. No significant effect was observed after IL-21 treatment.
Our data indicated that cytotoxicity method can be considered a low-cost alternative for NK cell isolation kits. It seems that culturing NK cells for 14 days in either PHA or OKT3 supplemented SCGM medium would be more effective than culturing for 16 days in the presence of IL-21.
鉴于近年来癌症死亡人数的增加以及常规疗法在根除癌症方面的不足，同时随着自然杀伤（NK）细胞疗法的发展，寻找新的有效抗癌疗法显得至关重要。乳腺癌是女性癌症死亡的第二大主要原因。由于大多数患者无法接受NK细胞疗法，因此尝试建立一种低成本且高效的方法来扩增和激活针对乳腺癌细胞系（MCF7）的NK细胞。
使用基于磁珠分选的NK细胞富集试剂盒或抗体及补体作为细胞毒性方法，从外周血单个核细胞（PBMCs）中分离NK细胞。然后，将NK细胞在含有饲养层（经辐照的PBMCs）的干细胞生长培养基（SCGM）中与PHA或OKT3一起培养。使用IL-2、IL-15和IL-21来扩增NK细胞，最后通过流式细胞术研究其细胞毒性活性。
获得了高纯度的NK细胞，两种分离方法之间未发现显著差异。使用IL-2加IL-15，16天后NK细胞数量增加了100倍。IL-21处理后未观察到显著效果。
我们的数据表明，细胞毒性方法可被视为NK细胞分离试剂盒的低成本替代方法。在添加PHA或OKT3的SCGM培养基中培养NK细胞14天似乎比在IL-21存在下培养16天更有效。
