Increasing 3D Matrix Rigidity Strengthens Proliferation and Spheroid Development of Human Liver Cells in a Constant Growth Factor Environment.
Mechanical forces influence the growth and shape of virtually all tissues and organs. Recent studies show that increased cell contractibility, growth and differentiation might be normalized by modulating cell tensions. Particularly, the role of these tensions applied by the extracellular matrix during liver fibrosis could influence the hepatocarcinogenesis process. The objective of this study is to determine if 3D stiffness could influence growth and phenotype of normal and transformed hepatocytes and to integrate extracellular matrix (ECM) stiffness to tensional homeostasis. We have developed an appropriate 3D culture model: hepatic cells within three-dimensional collagen matrices with varying rigidity. Our results demonstrate that the rigidity influenced the cell phenotype and induced spheroid clusters development whereas in soft matrices, Huh7 transformed cells were less proliferative, well-spread and flattened. We confirmed that ERK1 played a predominant role over ERK2 in cisplatin-induced death, whereas ERK2 mainly controlled proliferation. As compared to 2D culture, 3D cultures are associated with epithelial markers expression. Interestingly, proliferation of normal hepatocytes was also induced in rigid gels. Furthermore, biotransformation activities are increased in 3D gels, where CYP1A2 enzyme can be highly induced/activated in primary culture of human hepatocytes embedded in the matrix. In conclusion, we demonstrated that increasing 3D rigidity could promote proliferation and spheroid developments of liver cells demonstrating that 3D collagen gels are an attractive tool for studying rigidity-dependent homeostasis of the liver cells embedded in the matrix and should be privileged for both chronic toxicological and pharmacological drug screening.
Institut National de la Santé et de la Recherche Médicale (Inserm), UMR1085 Institut de Recherche sur la Santé l'Environnement et le Travail (IRSET); University of Rennes 1, SFR Biosit, F-35043, Rennes, France.
Jérémy
J
Bomo
Institut National de la Santé et de la Recherche Médicale (Inserm), UMR1085 Institut de Recherche sur la Santé l'Environnement et le Travail (IRSET); University of Rennes 1, SFR Biosit, F-35043, Rennes, France.
Frédéric
F
Ezan
Institut National de la Santé et de la Recherche Médicale (Inserm), UMR1085 Institut de Recherche sur la Santé l'Environnement et le Travail (IRSET); University of Rennes 1, SFR Biosit, F-35043, Rennes, France.
François
F
Tiaho
Institut National de la Santé et de la Recherche Médicale (Inserm), UMR1085 Institut de Recherche sur la Santé l'Environnement et le Travail (IRSET); University of Rennes 1, SFR Biosit, F-35043, Rennes, France.
Medjda
M
Bellamri
Institut National de la Santé et de la Recherche Médicale (Inserm), UMR1085 Institut de Recherche sur la Santé l'Environnement et le Travail (IRSET); University of Rennes 1, SFR Biosit, F-35043, Rennes, France.
Sophie
S
Langouët
Institut National de la Santé et de la Recherche Médicale (Inserm), UMR1085 Institut de Recherche sur la Santé l'Environnement et le Travail (IRSET); University of Rennes 1, SFR Biosit, F-35043, Rennes, France.
Nathalie
N
Theret
Institut National de la Santé et de la Recherche Médicale (Inserm), UMR1085 Institut de Recherche sur la Santé l'Environnement et le Travail (IRSET); University of Rennes 1, SFR Biosit, F-35043, Rennes, France.
Georges
G
Baffet
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D003094:Collagen / Q000737:chemistry; D003470:Culture Media / Q000737:chemistry*; D005782:Gels; D006244:Hardness; D022781:Hepatocytes / Q000502:physiology*; D006801:Humans; D008103:Liver Cirrhosis / Q000473:pathology; D020935:MAP Kinase Signaling System; D051381:Rats; D018874:Spheroids, Cellular / Q000502:physiology*
D003470:Culture Media; D005782:Gels; D003094:Collagen
BIOTRANSFORMATION; LIVER CELLS; MATRIX STIFFNESS; PROLIFERATION