A Combining Method to Enhance theIn VitroDifferentiation of Hepatic Precursor Cells

Abstract

The ideal bioartificial liver should be designed to reproduce as nearly as possible in vitro the habitat that hepatic cells find in vivo. In the present work, we investigated the in vitro perfusion condition with a view to improving the hepatic differentiation of pluripotent human liver stem cells (HLSCs) from adult liver. Tissue engineering strategies based on the cocultivation of HLSCs with hepatic stellate cells (ITO) and with several combinations of medium were applied to improve viability and differentiation. A mathematical model estimated the best flow rate for perfused cultures lasting up to 7 days. Morphological and functional assays were performed. Morphological analyses confirmed that a flow of perfusion medium (assured by the bioreactor system) enabled the in vitro organization of the cells into liver clusters even in the deeper levels of the sponge. Our results showed that, when cocultured with ITO using stem cell medium, HLSCs synthesized a large amount of albumin and the MTT test confirmed an improvement in cell proliferation. In conclusion, this study shows that our in vitro cell conditions promote the formation of clusters of HLSCs and enhance the functional differentiation into a mature hepatic population.

Treatment by Therapeutic Magnetic Resonance (TMR™) increases fibroblastic activity and keratinocyte differentiation in anin vitromodel of 3D artificial skin

“…Initially, the fibroblasts were seeded to allow for the production of the primary ECM molecules necessary for the proliferation and differentiation of endothelial cells. As reported in Figure a–d, and has been demonstrated in previous papers (Zavan et al , ; Carraro et al , ; Bressan et al ., ), fibroblasts secrete both structural and adhesive molecules, such as type I, III, IV and VII collagens, laminin‐1 and ‐5 and fibronectin, into the interstices of the 3D scaffolds.…”

MOL2NET 2015 - 1st International Conference on Synergies of Experimental Groups of Molecular and Biomedical Sciences with Data, Networks, and Social Sciences Experts. Bilbao,5–15 Dec, 2015.

“…It is proposed that hMSCs can play a key role in releasing trophic factors, which are important in cell protection and support following injury [25]. In our previous in vitro study, we have already demonstrated that hMSCs support hepatocytes maturation in a bioreactor system [26]. In this work, we additionally proved that also the HA-based scaffold plays a critical role in promoting the proliferation of endothelial cells, especially in the area surrounding the scaffold.…”

“…All 3D-cultures were maintained with DMEM supplemented with 10% FBS, 1% glutamine, and 10 nM dexamethasone in a bioreactor at 37 °C and 5% CO2. The bioreactor consisted in eight independent chambers, as previously described [26]. The seeded HA-based tubes were placed in the bioreactor, one in each separate chamber, where a perfusion medium flow rate of 0.5 mL/min through each scaffold was assured.…”

Ethnically diverse pluripotent stem cells for drug development

“…Evidence for the drug metabolizing activity of both Phase I and Phase II enzymes has been reported for pluripotent stem cell-generated hepatocyte-like cells [36,37,39,40,44,45,[47][48][49][50][51][52][53][54][55][56][57][58].…”