Scientific Research Center for Translational Medicine, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China; University of Chinese Academy of Sciences, Beijing 100049, China; School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China.
Scientific Research Center for Translational Medicine, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China.
Int J Biol Macromol. 2018 Apr 1;109:1154-1163. doi: 10.1016/j.ijbiomac.2017.11.103. Epub 2017 Nov 20.
Developing reliable three-dimensional (3D) cell culture systems that can mimic native tumor microenvironments is necessary for investigating the mechanism of hepatocellular carcinoma (HCC) metastasis and screen therapeutic drugs. In the present study, we developed decellularized liver matrix-alginate (DLM-ALG) hybrid gel beads. DLM powder was prepared by optimized decellularization methods and liquid nitrogen grinding. DLM-ALG beads were generated by dropping alginate solution containing DLM powder into a gelling bath. DLM powder concentration in alginate solution was ≤1% (w/v) and had no effect on the sphericity and mechanical stability of the beads. In addition, HCCLM3 cells cultured in 1% (w/v) DLM-ALG beads presented gradually enhanced viability during in vitro culture. The protein expression of urokinase plasminogen activator system and activity of matrix metalloproteinases (MMPs) of HCCLM3 cells, including MMP2 and MMP9, were more significantly promoted in DLM-ALG beads compared with that in conventional ALG beads without DLM powder. Moreover, the dose-dependent increase in HCCLM3 cell MMP activities was observed along with the DLM powder concentration in 0.5% and 1% DLM-ALG groups. Therefore, DLM-ALG beads might serve as a novel 3D culture system for exploring the mechanisms of HCC metastasis and screening therapeutic drugs.
开发能够模拟天然肿瘤微环境的可靠的三维（3D）细胞培养系统对于研究肝细胞癌（HCC）转移的机制和筛选治疗药物是必要的。在本研究中，我们开发了去细胞化的肝基质-藻酸盐（DLM-ALG）混合凝胶珠。通过优化的去细胞化方法和液氮研磨制备 DLM 粉末。通过将含有 DLM 粉末的藻酸盐溶液滴入凝胶浴中生成 DLM-ALG 珠。藻酸盐溶液中 DLM 粉末的浓度≤1%（w/v），并且对珠的球形度和机械稳定性没有影响。此外，在体外培养过程中，培养于 1%（w/v）DLM-ALG 珠中的 HCCLM3 细胞的活力逐渐增强。与不含 DLM 粉末的常规 ALG 珠相比，HCCLM3 细胞中尿激酶纤溶酶原激活系统的蛋白表达和基质金属蛋白酶（MMPs）的活性，包括 MMP2 和 MMP9，在 DLM-ALG 珠中得到了更显著的促进。此外，随着 0.5%和 1%DLM-ALG 组中 DLM 粉末浓度的增加，HCCLM3 细胞 MMP 活性呈剂量依赖性增加。因此，DLM-ALG 珠可能成为探索 HCC 转移机制和筛选治疗药物的新型 3D 培养系统。
