Laura Falasca
Sapienza University of Rome
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Featured researches published by Laura Falasca.
Cells Tissues Organs | 2001
Laura Falasca; Alfredo Miccheli; Elisabetta Sartori; Alberta Tomassini; Laura Conti Devirgiliis
The maintenance of the differentiated hepatocyte phenotype and its specific physiological properties is known to depend on several factors, such as chemical signals, cell-cell and extracellular matrix molecular interactions, as well as the use of three-dimensional matrices. The entrapment of hepatocytes within Ca-alginate at high cell density and the culture under continuous flow favour the development of three-dimensional organization and promote expression of the differentiated hepatic phenotype. This system could represent an improvement in hepatocyte cultivation for basic studies of liver physiology and metabolism; it could also be applicable in toxicology, hepatocyte transplantation or development of bioartificial organs. This report describes the effect of alginate entrapment and culture in a bioreactor on hepatocyte aggregate formation, with particular attention to the re-establishment of cell polarity, cell junctions and three-dimensional re-organization of the cytoskeleton. Oxygen supply and cell oxygen consumption rate were monitored in order to evaluate possible changes in hepatocyte energy requirement. Our data show that after only 6 h of perfusion in the bioreactor, actin and cytokeratin localize along the adhesion areas of the plasma membrane, in which reconstituted bile canaliculi were also observed. Moreover, the presence of connexin at the level of joined membranes of neighbouring cells suggests the establishment of gap junctions between hepatocytes. After the first 30 min of perfusion the oxygen consumption rate remained constant throughout the experimental period.
Cell and Tissue Research | 1998
Laura Falasca; Anna Favale; Annalucia Serafino; Carmen Ara; Laura Conti Devirgiliis
Abstract The usefulness of cultured hepatocytes is limited by the gradual loss of their typical physiological functions that occurs in vitro, mainly due to the absence of microenviromental conditions found in vivo. In this study we describe the effect of retinoic acid on the re-establishment of morphological characteristics and on the reorganization of the cytoskeletal network in cultured rat hepatocytes. Results obtained demonstrate that retinoic acid can influence hepatocyte differentiation, as regards the recovery of cell polarity, polyhedric shape and reformation of bile canaliculi and junctional complexes. The main target of this action appears to be the cytoarchitecture of cytoskeletal components, particularly cytokeratin filaments, which regain the configuration present in intact liver. The reorganization of the intermediate filaments does not seem to be dependent on the induction of higher levels of cytokeratin proteins, but rather appears to be due to post-translational regulation. The effect of retinoic acid on the cytoskeletal organization could determine the stabilization of intercellular contacts by means of junctions, leading to the appearance of morpho-functional characteristics typical of well-differentiated hepatocytes.
Cytotechnology | 2000
Alfredo Miccheli; Alberta Tomassini; G. Capuani; Maria Enrica Di Cocco; Elisabetta Sartori; Laura Falasca; Laura Conti Devirgiliis; Cesare Manetti; Filippo Conti
We studied the effect of continuous medium flow on the viabilityand structural organization of hepatocytes high density entrapped inalginate gel beads in the first few hours after isolation.The metabolic energy status of the entrapped cells, monitored invivo by 31P NMR spectroscopy, was stable during theexperimental time and a physiological redox ratio was reachedafter the first three hours of culture. The morphologicalanalysis revealed that the entrapped hepatocytes placed in a fixed-bed bioreactor under continuous flow showed a polyhedricalshape with numerous microvilli on cell surface and reconstitutedtight junctions as well as bile canalicular structures, closelyresembling those present in the liver.These results suggest that continuous flow allows the culture ofhepatocytes at very high cell density within a matrix withoutloss of viability and accelerates cellular tissue reconstructionat very short times after isolation. This type of culture couldrepresent a very useful model for physiological andtoxicological studies as well as a promising approach toward thedevelopment of a bioartificial hybrid support device in acuteliver failure.
Bioscience Reports | 1996
Mara Massimi; Laura Falasca; Angelina Felici; Luciana Dini; L. Conti Devirgiliis
The influence of cell density on expression of the asialoglycoprotein receptor system in primary cultures of rat hepatocytes was evaluated by measuring the level of the receptor specific mRNA. When the hepatocytes are cultured at high cellular density and are not in a proliferative condition, the transcript molecules of the receptor appear increased about 50% with respect to the low plating density, indicating a modulation of asialoglycoprotein receptor expression at transcriptional level. Such control may be dependent on surface molecules involved in cell specific reassociation, since it is well known that cell contacts play a significant regulatory role in differentiated cells.
Artificial Cells, Blood Substitutes, and Biotechnology | 2000
G. Capuani; A. Miccheli; Alberta Tomassini; Laura Falasca; Tommaso Aureli; Filippo Conti
Cellular volume of hepatocytes entrapped in alginate gel beads were evaluated under in vivo conditions in samples having different cell densities by applying matemathical models to the diffusion data obtained by magnetic resonance imaging (MRI). The calculated average volume is in good agrement with the values from the literature - being closer to the data relative to living tissue than to isolated cells. The non invasive characteristics of magnetic resonance imaging make this method particularly well suited to obtain information from the intact system.
Mechanisms of Ageing and Development | 2001
Laura Falasca; Angelina Felici; Mara Massimi; Luciana Dini; Laura Conti Devirgiliis
The influence of retinoic acid on the expression of a typical marker of hepatocyte differentiation, i.e. the asialoglycoprotein receptor, has been studied. Cultured hepatocytes, isolated from adult rats, a model of quiescent, mature cells and from 20-day-old fetuses, a model of proliferating and less differentiated cells, were used. The asialoglycoprotein receptor expression appears to be affected by retinoic acid during prenatal life; both mRNA level and protein amount increased in fetal hepatocytes, but no modification has been found in adult cells, suggesting a regulative effect of retinoic acid during prenatal life, acting at transcriptional and/or translational level. Surprisingly, the receptor binding activity of adult hepatocytes is decreased after retinoic acid treatment, indicating a possible further modulation by this molecule on receptor activity at the post-translational level.
Hepatology | 1998
Laura Falasca; Anna Favale; Giampiero Gualandi; Gennaro Maietta; Laura Conti Devirgiliis
European Journal of Cell Biology | 1993
Luciana Dini; Laura Falasca; Alessandro Lentini; P Mattioli; Mauro Piacentini; L Piredda; F. Autuori
Anticancer Research | 1999
Laura Falasca; Pierina Marcellini; Carmen Ara; Anna Rufo; Laura Conti Devirgiliis
Cellular and Molecular Biology | 1996
Luciana Dini; Laura Falasca; L Rossi; G. Rotilio