Abstract
Background aims. Previously, we have shown that human decidua-derived mesenchymal stromal cells (DMSC) are mesenchymal stromal cells (MSC) with a clonal differentiation capacity for the three embryonic layers. The endodermal capacity of DMSC was revealed by differentiation into pulmonary cells. In this study, we examined the hepatic differentiation of DMSC. Methods. DMSC were cultured in hepatic differentiation media or co-cultured with murine liver homogenate and analyzed with phenotypic, molecular and functional tests. Results and Conclusions. DMSC in hepatic differentiation media changed their fibroblast morphology to a hepatocyte-like morphology and later formed a 3-dimensional (3-D) structure or hepatosphere. Moreover, the hepatocyte-like cells and the hepatospheres expressed liver-specific markers such as synthesis of albumin (ALB), hepatocyte growth factor receptor (HGFR), α-fetoprotein (AFP) and cytokeratin-18 (CK-18), and exhibited hepatic functions including glycogen storage capacity and indocyanine green (ICG) uptake/secretion. Human DMSC co-cultured with murine liver tissue homogenate in a non-contact in vitro system showed hepatic differentiation, as evidenced by expression of AFP and ALB genes. The switch in the expression of these two genes resembled liver development. Indeed, the decrease in AFP and increase in ALB expression throughout the co-culture were consistent with the expression pattern observed during normal liver organogenesis in the embryo. Interestingly, AFP and ALB expression was significantly higher when DMSC were co-cultured with injured liver tissue, indicating that DMSC respond differently under normal and pathologic micro-environmental conditions. In conclusion, DMSC-derived hepatospheres and DMSC co-cultured with liver homogenate could be suitable in vitro models for toxicologic, developmental and pre-clinical hepatic regeneration studies.
Acknowledgements
The authors thank to Dr I. Vegh for critical reading of the manuscript and English grammar revision, and to the veterinarian Montserrat Grau for her assistance with the animal protocols (Research Centre, Research Institute Hospital 12 de Octubre). We also thank to J. C. Gonzalez for his help with the confocal microscopy analysis and J. C. Rubio for his contribution to the real time PCR determinations.
Disclosure of interests: the authors declare that no conflict of interests exists. This work was supported by grants to Dr Flores from the Spanish Institute of Health Carlos III (FIS 05/2288; CP03/00004), the MMA Foundation (2004-029; 2005-088), the Neurosciences and Aging Foundation, and a contract associated with Dr Flores’ project 05/2288 from the Hospital 12 Octubre Biomedical Research Foundation.