Concurrent isolation of hepatic stem cells and hepatocytes from the human liver

Hepatocytes differentiated from induced pluripotent stem cells or stem cells have the potential to be representative in vitro models of the human liver for research as well as early safety assessment programs. However, up until now, there has been no definitive proof that differentiated hepatocytes recapitulate the phenotype and functional characteristics of primary hepatocytes from the same individual. Thus, a method for the concurrent isolation of hepatocytes and hepatic stem cells is presented here to provide the cells necessary for the evaluation of the required benchmarking. The method presented here generated high-quality hepatocytes with a purity of 94 ± 1% and a high percentage viability of 79 ± 2%. Furthermore, the hepatic stem cells isolated were found to be actively proliferating and have a purity of 98 ± 1%. Thus, these isolated cells can be used as a powerful tool for the validation of differentiated hepatocyte in vitro models

Evidence of Potential Averaging over the Finite Surface of a Bioelectric Surface Electrode

Most bioelectric signals are not only functions of time but also exhibit a variation in spatial distribution. Surface EMG signals are often "summarized" by a large electrode. The effect of such an electrode is interpreted as averaging the potential at the surface of the skin beneath the electrode. We first introduce an electrical equivalent model to delineate this principle of averaging. Next, in a realistic finite element model of EMG generation, two outcome variables are evaluated to assess the validity of the averaging principle. One is the change in voltage distribution in the volume conductor after electrode application. The other is the change in voltage across the high impedance double layer between tissue and electrode. We found that the principle of averaging is valid, once the impedance of the double layer is sufficiently high. The simulations also revealed that skin conductivity plays a role. High-density surface EMG provided experimental evidence consistent with the simulation results. A grid with 120 small electrodes was placed over the thenar muscles of the hand. Electrical nerve stimulation assured a reproducible compound muscle response. The averaged grid response was compared with a single electrode matching the surface of the high-density electrodes. The experimental results showed relatively small errors indicating that averaging of the surface potential by the electrode is a valid principle under most practical conditions. © 2009 Biomedical Engineering Society

Using two-dimensional spatial information in decomposition of surface EMG signals.

Contains fulltext : 51651.pdf (publisher's version ) (Closed access)Recently, high-density surface EMG electrode grids and multi-channel amplifiers became available for non-invasive recording of human motor units (MUs). We present a way to decompose surface EMG signals into MU firing patterns, whereby we concentrate on the importance of two-dimensional spatial differences between the MU action potentials (MUAPs). Our method is exemplified with high-density EMG data from the vastus lateralis muscle of a single subject. Bipolar and Laplacian spatial filtering was applied to the monopolar raw signals. From the single recording in this subject six different simultaneously active MUs could be distinguished using the spatial differences between MUAPs in the direction perpendicular to the muscle fiber direction. After spike-triggered averaging, 125-channel two-dimensional MUAP templates were obtained. Template-matching allowed tracking of all MU firings. The impact of spatial information was measured by using subsets of the MUAP templates, either in parallel or perpendicular to the muscle fiber direction. The use of one-dimensional spatial information perpendicular to the muscle fiber direction was superior to the use of a linear array electrode in the longitudinal direction. However, to detect the firing events of the MUs with a high accuracy, as needed for instance for estimation of firing synchrony, two-dimensional information from the complete grid electrode appears essential

Characterization of a long-term mouse primary liver 3D tissue model recapitulating innate-immune responses and drug-induced liver toxicity

Three-dimensional liver in vitro systems have recently attracted a lot of attention in drug development. These systems help to gain unprecedented insights into drug-induced liver injury (DILI), as they more closely reproduce liver biology, and as drug effects can be studied in isolated and controllable microenvironments. Many groups established human-based in vitro models but so far neglected the animal equivalent, although the availability of both models would be desirable. Animal in vitro models enable back- and forward translation of in vitro and in vivo findings, bridge the gap between rodent in vivo and human in vitro scenarios, and ultimately support the interpretation of data generated with preclinical species and humans. Since mice are often used in drug development and physiologically relevant in vitro systems are lacking, we established, for the first time, a mouse liver model that encompasses primary parenchymal and non-parenchymal cells with preserved viability and functionality over three weeks. Using our three-dimensional liver spheroids, we were able to predict the toxicity of known DILI compounds, demonstrated the interaction cascades between the different cell types and showed evidence of drug-induced steatosis and cholestasis. In summary, our mouse liver spheroids represent a valuable in vitro model that can be applied to study DILI findings, reported from mouse studies, and offers the potential to detect immune-mediated drug-induced liver toxicity.ISSN:1932-620

Characterization of a long-term mouse primary liver 3D tissue model recapitulating innate-immune responses and drug-induced liver toxicity.

Three-dimensional liver in vitro systems have recently attracted a lot of attention in drug development. These systems help to gain unprecedented insights into drug-induced liver injury (DILI), as they more closely reproduce liver biology, and as drug effects can be studied in isolated and controllable microenvironments. Many groups established human-based in vitro models but so far neglected the animal equivalent, although the availability of both models would be desirable. Animal in vitro models enable back- and forward translation of in vitro and in vivo findings, bridge the gap between rodent in vivo and human in vitro scenarios, and ultimately support the interpretation of data generated with preclinical species and humans. Since mice are often used in drug development and physiologically relevant in vitro systems are lacking, we established, for the first time, a mouse liver model that encompasses primary parenchymal and non-parenchymal cells with preserved viability and functionality over three weeks. Using our three-dimensional liver spheroids, we were able to predict the toxicity of known DILI compounds, demonstrated the interaction cascades between the different cell types and showed evidence of drug-induced steatosis and cholestasis. In summary, our mouse liver spheroids represent a valuable in vitro model that can be applied to study DILI findings, reported from mouse studies, and offers the potential to detect immune-mediated drug-induced liver toxicity

Trueness of full-arch IO scans estimated based on 3D translational and rotational deviations of single teeth—an in vitro study

Objectives!#!To three-dimensionally evaluate deviations of full-arch intraoral (IO) scans from reference desktop scans in terms of translations and rotations of individual teeth and different types of (mal)occlusion.!##!Materials and methods!#!Three resin model pairs reflecting different tooth (mal)positions were mounted in the phantom head of a dental simulation unit and scanned by three dentists and three non-graduate investigators using a confocal laser IO scanner (Trios 3®). The tooth-crown surfaces of the IO scans and reference scans were superimposed by means of best-fit alignment. A novel method comprising the measurement of individual tooth positions was used to determine the deviations of each tooth in the six degrees of freedom, i.e., in terms of 3D translation and rotation. Deviations between IO and reference scans, among tooth-(mal)position models, and between dentists and non-graduate investigators were analyzed using linear mixed-effects models.!##!Results!#!The overall translational deviations of individual teeth on the IO scans were 76, 32, and 58 µm in the lingual, mesial, and intrusive directions, respectively, resulting in a total displacement of 114 µm. Corresponding rotational deviations were 0.58° buccal tipping, 0.04° mesial tipping, and 0.14° distorotation leading to a combined rotation of 0.78°. These deviations were the smallest for the dental arches with anterior crowding, followed by those with spacing and those with good alignment (p < 0.05). Results were independent of the operator's level of education.!##!Conclusions!#!Compared to reference desktop scans, individual teeth on full-arch IO scans showed high trueness with total translational and rotational deviations < 115 µm and < 0.80°, respectively.!##!Clinical relevance!#!Available confocal laser IO scanners appear sufficiently accurate for diagnostic and therapeutic orthodontic applications. Results indicate that full-arch IO scanning can be delegated to non-graduate dental staff members