Laboratory of Biomaterials, Institute for Life and Medical Sciences, Kyoto University, 53 Kawara-cho Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
ACS Biomater Sci Eng. 2023 May 8;9(5):2672-2682. doi: 10.1021/acsbiomaterials.2c01420. Epub 2023 Apr 4.
As one imaging method to evaluate monocyte-macrophage differentiation, cationized gelatin nanospheres incorporating a molecular beacon (MB) (cGNS) were designed. Cationized gelatin nanospheres (cGNS) of different apparent sizes were prepared by the conventional coacervation method, and then the MB of CD204 was incorporated into cGNS to prepare cGNS. When three types of cGNS were cultured with human monocytoma (THP-1) cells, the cGNS with a 110 nm diameter showed the highest MB delivery efficiency. In addition, no influence on the monocyte-macrophage differentiation was observed in terms of CD204 gene expression and cell viability. After incubation with cGNS incorporating CD204 MB (cGNS), THP-1 cells were stimulated by phorbol 12-myristate 13-acetate (PMA) for monocyte differentiation into macrophages. The fluorescence intensity of macrophages increased with the incubation time. In contrast, the fluorescence intensity of macrophages incubated with MB alone was not changed. On the other hand, there was no change in the fluorescence intensity of original THP-1 cells cultured with cGNS. It is concluded that the cGNS are promising to trace the differentiation of THP-1 cells into macrophages in their live condition.
作为评估单核细胞-巨噬细胞分化的一种成像方法，设计了结合分子信标的阳离子化明胶纳米球（cGNS）。通过常规凝聚法制备了不同表观尺寸的阳离子化明胶纳米球（cGNS），然后将 CD204 的分子信标（MB）掺入 cGNS 中制备 cGNS。当三种类型的 cGNS 与人类单核细胞瘤（THP-1）细胞共培养时，直径为 110nm 的 cGNS 显示出最高的 MB 传递效率。此外，CD204 基因表达和细胞活力方面没有观察到对单核细胞-巨噬细胞分化的影响。用 CD204 MB 结合的 cGNS（cGNS）孵育 THP-1 细胞后，用佛波醇 12-肉豆蔻酸 13-醋酸盐（PMA）刺激单核细胞分化为巨噬细胞。巨噬细胞的荧光强度随孵育时间的增加而增加。相比之下，单独用 MB 孵育的巨噬细胞的荧光强度没有变化。另一方面，用 cGNS 培养的原始 THP-1 细胞的荧光强度没有变化。结论是，cGNS 有望在活细胞条件下追踪 THP-1 细胞向巨噬细胞的分化。
