Novel decellularized liver matrix-alginate hybrid gel beads for the 3D culture of hepatocellular carcinoma cells

• Published in: International journal of biological macromolecules Vol. 109; pp. 1154 - 1163
• Main Authors: Sun, Dongsheng, Liu, Yang, Wang, Huihui, Deng, Fei, Zhang, Ying, Zhao, Shan, Ma, Xiaojun, Wu, Huijian, Sun, Guangwei
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2018
• Subjects: Alginate gel bead; alginates; cell culture; Decellularized liver matrix; dose response; drugs; gelation; gels; grinding; Hepatocellular carcinoma; hepatoma; liquids; liver; Matrix metalloproteinase; metalloproteinases; metastasis; neoplasm cells; nitrogen; protein synthesis; screening; Three-dimensional (3D) cell culture; u-plasminogen activator; Urokinase plasminogen activator; viability; DLM-ALG beads; Urokinase plasminogen activator; DLM; tPA; H&E; uPA; Three-dimensional (3D) cell culture; HCC; Hepatocellular carcinoma; ALG beads; Matrix metalloproteinase; ECM; MMP; phospho-ERK (p-ERK); Decellularized liver matrix; ESEM; PAI-1; Alginate gel bead; TIMP; ERK
• ISSN: 0141-8130; 1879-0003; 1879-0003
• DOI: 10.1016/j.ijbiomac.2017.11.103

•DLM–ALG beads were developed to mimic native hepatocellular carcinoma microenvironments.•The invasion and metastasis potential of HCC cells in DLM–ALG beads was enhanced than that in conventional ALG beads.•DLM–ALG beads might serve as a novel in vitro 3D culture system for HCC research. 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.