Generation of Multilayered 3D Structures of HepG2 Cells Using a Bio-printing Technique
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Background/Aims: Chronic liver disease is a major widespread cause of death, and whole liver transplantation is the only definitive treatment for patients with end-stage liver diseases. However, many problems, including donor shortage, surgical complications and cost, hinder their usage. Recently, tissue-engineering technology provided a potential breakthrough for solving these problems. Three-dimensional (3D) printing technology has been used to mimic tissues and organs suitable for transplantation, but applications for the liver have been rare. Methods: A 3D bioprinting system was used to construct 3D printed hepatic structures using alginate. HepG2 cells were cultured on these 3D structures for 3 weeks and examined by fluorescence microscopy, histology and immunohistochemistry. The expression of liverspecific markers was quantified on days 1, 7, 14, and 21. Results: The cells grew well on the alginate scaffold, and liver-specific gene expression increased. The cells grew more extensively in 3D culture than two-dimensional culture and exhibited better structural aspects of the liver, indicating that the 3D bioprinting method recapitulates the liver architecture. Conclusions: The 3D bioprinting of hepatic structures appears feasible. This technology may become a major tool and provide a bridge between basic science and the clinical challenges for regenerative medicine of the liver.
Abstract
Background/Aims: Chronic liver disease is a major widespread cause of death, and whole liver transplantation is the only definitive treatment for patients with end-stage liver diseases. However, many problems, including donor shortage, surgical complications and cost, hinder their usage. Recently, tissue-engineering technology provided a potential breakthrough for solving these problems. Three-dimensional (3D) printing technology has been used to mimic tissues and organs suitable for transplantation, but applications for the liver have been rare. Methods: A 3D bioprinting system was used to construct 3D printed hepatic structures using alginate. HepG2 cells were cultured on these 3D structures for 3 weeks and examined by fluorescence microscopy, histology and immunohistochemistry. The expression of liverspecific markers was quantified on days 1, 7, 14, and 21. Results: The cells grew well on the alginate scaffold, and liver-specific gene expression increased. The cells grew more extensively in 3D culture than two-dimensional culture and exhibited better structural aspects of the liver, indicating that the 3D bioprinting method recapitulates the liver architecture. Conclusions: The 3D bioprinting of hepatic structures appears feasible. This technology may become a major tool and provide a bridge between basic science and the clinical challenges for regenerative medicine of the liver.
참고문헌 (34)
- Agarwal S, "Efficient differentiation of functional hepatocytes from human embryonic stem cells", Stem Cells, 26, pp.1117~1127 (2008)
- Sancho-Bru P, "Directed differentiation of murine-induced pluripotent stem cells to functional hepatocytelike cells", J Hepatol, 54, pp.98~107 (2011)
- Shiraki N, "Differentiation of mouse and human embryonic stem cells into hepatic lineages", Genes Cells, 13, pp.731~746 (2008)
- Sullivan GJ, "Generation of functional human hepatic endoderm from human induced pluripotent stem cells", Hepatology, 51, pp.329~335 (2010)
- Basma H, "Differentiation and transplantation of human embryonic stem cell-derived hepatocytes", Gastroenterology, 136, pp.990~999 (2009)
- Duan Y, "Differentiation and enrichment of hepatocyte-like cells from human embryonic stem cells in vitro and in vivo", Stem Cells, 25, pp.3058~3068 (2007)
- Gai H, "Generation of murine hepatic lineage cells from induced pluripotent stem cells", Differentiation, 79, pp.171~181 (2010)
- Touboul T, "Generation of functional hepatocytes from human embryonic stem cells under chemically defined conditions that recapitulate liver development", Hepatology, 51, pp.1754~1765 (2010)
- He ZY, "Murine embryonic stem cell-derived hepatocytes correct metabolic liver disease after serial liver repopulation", Int J Biochem Cell Biol, 44, pp.648~658 (2012)
- Soto-Gutiérrez A, "Reversal of mouse hepatic failure using an implanted liver-assist device containing ES cell-derived hepatocytes", Nat Biotechnol, 24, pp.1412~1419 (2006)
- Teratani T, "Direct hepatic fate specification from mouse embryonic stem cells", Hepatology, 41, pp.836~846 (2005)
- Baharvand H, "Differentiation of human embryonic stem cells into hepatocytes in 2D and 3D culture systems in vitro", Int J Dev Biol, 50, pp.645~652 (2006)
- Gieseck RL, "Hannan NR, Bort R, et al. Maturation of induced pluripotent stem cell derived hepatocytes by 3D-culture", PLoS One, 9, pp.e86372~ (2014)
- Godoy P, "Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME", Arch Toxicol, 87, pp.1315~1530 (2013)
- Mironov V, "Organ printing: computer-aided jet-based 3D tissue engineering", Trends Biotechnol, 21, pp.157~161 (2003)
- Williams DF, "On the mechanisms of biocompatibility", Biomaterials, 29, pp.2941~2953 (2008)
- Bailey MJ, "The use of solid physical models for the study of macromolecular assembly", Curr Opin Struct Biol, 8, pp.202~208 (1998)
- Shimizu TS, "Molecular model of a lattice of signalling proteins involved in bacterial chemotaxis", Nat Cell Biol, 2, pp.792~796 (2000)
- Symes MD, "Integrated 3D-printed reactionware for chemical synthesis and analysis", Nat Chem, 4, pp.349~354 (2012)
- Chia HN, "Recent advances in 3D printing of biomaterials", J Biol Eng, 9, pp.4~ (2015)
- Jones N, "Science in three dimensions: the print revolution", Nature, 487, pp.22~23 (2012)
- Murphy SV, "3D bioprinting of tissues and organs", Nat Biotechnol, 32, pp.773~785 (2014)
- Dhawan A, "Hepatocyte transplantation for liver-based metabolic disorders", J Inherit Metab Dis, 29, pp.431~434 (2006)
- Du Y, "Human hepatocytes with drug metabolic function induced from fibroblasts by lineage reprogramming", Cell Stem Cell, 14, pp.394~403 (2014)
- Huang P, "Induction of functional hepatocytelike cells from mouse fibroblasts by defined factors", Nature, 475, pp.386~389 (2011)
- Huang P, "Direct reprogramming of human fibroblasts to functional and expandable hepatocytes", Cell Stem Cell, 14, pp.370~384 (2014)
- Sekiya S, "Direct conversion of mouse fibroblasts to hepatocyte-like cells by defined factors", Nature, 475, pp.390~393 (2011)
- Boland T, "Cell and organ printing 2: fusion of cell aggregates in three-dimensional gels", Anat Rec A Discov Mol Cell Evol Biol, 272, pp.497~502 (2003)
- Huh D, "Ingber DE. Reconstituting organ-level lung functions on a chip", Science, 328, pp.1662~1668 (2010)
- Sonntag F, "Design and prototyping of a chip-based multi-micro-organoid culture system for substance testing, predictive to human (substance) exposure", J Biotechnol, 148, pp.70~75 (2010)
- Duan B, "3D bioprinting of heterogeneous aortic valve conduits with alginate", gelatin hydrogels, 2013, pp.101, 1255~1264
- Jakab K, "Tissue engineering by selfassembly and bio-printing of living cells", Biofabrication, 2, pp.022001~ (2010)
- Marga F, "Toward engineering functional organ modules by additive manufacturing", Biofabrication, 4, pp.022001~ (2012)
- Kwon YJ, "Clinical implications of advances in liver regeneration", Clin Mol Hepatol, 21, pp.7~13 (2015)