Layered Long Term Co-Culture of Hepatocytes and Endothelial Cells on a Transwell Membrane: Toward Engineering the Liver Sinusoid

This paper presents a novel liver model that mimics the liver sinusoid where most liver activities occur. A key aspect of our current liver model is a layered co-culture of primary rat hepatocytes (PRHs) and primary rat liver sinusoidal endothelial cells (LSECs) or bovine aortic endothelial cells (BAECs) on a transwell membrane. When a layered co-culture was attempted with a thin matrigel layer placed between hepatocytes and endothelial cells to mimic the Space of Disse, the cells did not form completely separated monolayers. However, when hepatocytes and endothelial cells were cultured on the opposite sides of a transwell membrane, PRHs co-cultured with LSECs or BAECs maintained their viability and normal morphology for 39 and 57 days, respectively. We assessed the presence of hepatocyte-specific differentiation markers to verify that PRHs remained differentiated in the long-term co-culture and analyzed hepatocyte function by monitoring urea synthesis. We also noted that the expression of cytochrome P-450 remained similar in the cocultured system from Day 13 to Day 48. Thus, our novel liver model system demonstrated that primary hepatocytes can be cultured for extended times and retain their hepatocyte-specific functions when layered with endothelial cells

Flow-Induced Voltage Generation Over Monolayer Graphene in the Presence of Herringbone Grooves

While flow-induced voltage over a graphene layer has been reported, its origin remains unclear. In our previous study, we suggested different mechanisms for different experimental configurations: phonon dragging effect for the parallel alignment and an enhanced out-of-plane phonon mode for the perpendicular alignment (Appl. Phys. Lett. 102:063116, 2011). In order to further examine the origin of flow-induced voltage, we introduced a transverse flow component by integrating staggered herringbone grooves in the microchannel. We found that the flow-induced voltage decreased significantly in the presence of herringbone grooves in both parallel and perpendicular alignments. These results support our previous interpretation

The Physical Foundation of Vasoocclusion in Sickle Cell Disease

AbstractThe pathology of sickle cell disease arises from the occlusion of small blood vessels because of polymerization of the sickle hemoglobin within the red cells. We present measurements using a microfluidic method we have developed to determine the pressure required to eject individual red cells from a capillary-sized channel after the cell has sickled. We find that the maximum pressure is only ∼100 Pa, much smaller than typically found in the microcirculation. This explains why experiments using animal models have not observed occlusion beginning in capillaries. The magnitude of the pressure and its dependence on intracellular concentration are both well described as consequences of sickle hemoglobin polymerization acting as a Brownian ratchet. Given the recently determined stiffness of sickle hemoglobin gels, the observed obstruction seen in sickle cell disease as mediated by adherent cells can now be rationalized, and surprisingly suggests a window of maximum vulnerability during circulation of sickle cells

Layered Hepatocytes and Endothelial Cells on a Transwell Membrane: Toward Engineering the Liver Sinusoid

This paper presents a novel liver model that mimics the liver sinusoid where most liver activities occur. A key aspect of our current liver model is a layered co-culture of primary rat hepatocytes (PRHs) and primary rat liver sinusoidal endothelial cells (LSECs) or bovine aortic endothelial cells (BAECs) on a transwell membrane. When a layered co-culture was attempted with a thin Matrigel layer placed between hepatocytes and endothelial cells to mimic the space of Disse, the cells did not form completely separated monolayers. However, when hepatocytes and endothelial cells were cultured on the opposite sides of a transwell membrane, PRHs co-cultured with LSECs or BAECs maintained their viability and normal morphology for 39 and 57 days, respectively. We assessed the presence of hepatocyte-specific differentiation markers to verify that PRHs remained differentiated in the long-term co-culture and analyzed hepatocyte function by monitoring urea synthesis. We also noted that the expression of cytochrome P-450 remained similar in the co-cultured system from day 1 to day 48. Thus, our novel liver model system demonstrated that primary hepatocytes can be cultured for extended times and retain their hepatocyte-specific functions when layered with endothelial cells

Magnetically actuated micropumps using an Fe-PDMS composite membrane

Proceedings SPIE International Society for Optical Engineering 6172 (2006). Retrieved April 2006 from http://mems.mem.drexel.edu/actuator.pdfIn this paper we describe a novel Fe-PDMS composite that can be used to create magnetically actuated polymeric microstructures. The composite is formed by suspending <10μm iron particles in polydimethylsiloxane (PDMS) at concentrations ranging from 25-75% by weight. Material properties and processing capabilities have been examined, and to demonstrate the usefulness of this material we have designed, fabricated and tested two prototypical micropumps that utilize an Fe-PDMS actuator membrane

Parylene Microcolumn for Miniature Gas Chromatograph

This research contributes to worldwide efforts to miniaturize one of the most powerful and versatile analytical tools, gas chromatography (GC). If a rapid, sensitive and selective hand-held GC system is realized, it would have a wide range of applications in many industries and research areas. As a part of developing a hand-held GC system, this research focuses on the separation column, which is the most important component of a GC system. This thesis describes the development of a miniature separation column that has low thermal mass and an embedded heating element for rapid thermal cycling. The worlds first thin polymer film (parylene) GC column has been successfully developed. This thesis includes: first, a study of theoretical column performance of rectangular GC column; second, the design optimization of parylene column and embedded heating element; third, the development of new processes such as parylene micromolding and stationary phase coating technique for parylene column; fourth, the fabrication of parylene GC column with an embedded heating element; and lastly, the testing and evaluation of parylene GC column through GC analysis.Ph.D.Committee Chair: Peter J. Hesketh; Committee Member: Boris Mizaikoff; Committee Member: Bruno Frazier; Committee Member: Wenjing Ye; Committee Member: Yogendra Josh

AC Electrokinetic Phenomena Generated by Microelectrode Structures

The field of AC electrokinetics is rapidly growing due to its ability to perform dynamic fluid and particle manipulation on the micro- and nano-scale, which is essential for Lab-on-a-Chip applications. AC electrokinetic phenomena use electric fields to generate forces that act on fluids or suspended particles (including those made of dielectric or biological material) and cause them to move in astonishing ways1, 2. Within a single channel, AC electrokinetics can accomplish many essential on-chip operations such as active micro-mixing, particle separation, particle positioning and micro-pattering. A single device may accomplish several of those operations by simply adjusting operating parameters such as frequency or amplitude of the applied voltage. Suitable electric fields can be readily created by micro-electrodes integrated into microchannels. It is clear from the tremendous growth in this field that AC electrokinetics will likely have a profound effect on healthcare diagnostics3-5, environmental monitoring6 and homeland security7