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At least 775 records · Page 43Linked to original sources
PubMed1986

Cellular proliferation in the vitreous: the use of vitreous explants as a model system.

An in vitro model of cellular proliferation in the vitreous has been developed using explants of bovine vitreous gel. Various cell types, including chick embryo pigmented retinal epithelium, choroidal fibroblasts, retinal glial cells, bovine retinal capillary endothelial cells, and peritoneal mouse macrophages, were cultured at 37 degrees C on vitreous gels for periods up to 8 days and compared for their effects on the structure of the gel. Choroidal fibroblasts caused a very marked reduction in the size of the gel and produced the strongest traction on the gel fibril structure; in contrast, peritoneal macrophages caused virtually no reduction in vitreous volume and little or no visible traction on the gel as detected by phase-contrast microscopy. The remaining cell types usually formed sheets on the surface of the gel; this was associated with an initial reduction in vitreous volume of approximately 50%, but little change thereafter. The cell mediated traction events on the structure of the vitreous gel were followed by time lapse video microscopy and by electron microscopy at various times after seeding of cells on the gels. The appearances corresponded closely with reported clinico-pathological studies of cellular proliferation in the vitreous. We believe that this in vitro model has several advantages over in vivo models of cellular proliferation in the vitreous, in that it permits analysis of individual cell behaviour and it is eminently suitable for pharmacologic manipulation.

Animals↗
PubMed1998

Acousto-optic random-access laser scanning microscopy: fundamentals and applications to optical recording of neuronal activity.

A novel approach to laser scanning microscopy is presented that utilizes diffraction-based scanning principles to achieve fast random-access positioning of a focused laser beam. This non-imaging approach overcomes the speed limitation of present reflection-based scanning microscopes while maintaining high spatial resolution. The presented system combines conventional video microscopy with fast non-imaging scanning microscopy. Together with readily available optical indicators of neuronal activity, this system permits multi-site optical recording from living brain tissue. In this paper, we will review the underlying principles of laser scanning microscopy and the steps in development that led to the current acousto-optic scanning system. We will present typical signals recorded with the current system, and we will outline ongoing extensions of the system. We will also discuss the present limitation of this instrumentation and look into directions of future development.

Acoustics↗
PubMed1999

Dynamic in vivo measurement of erythrocyte velocity and flow in capillaries and of microvessel diameter in the rat brain by confocal laser microscopy.

A new method for studying brain microcirculation is described. Both fluorescently labeled erythrocytes and plasma were visualized on-line through a closed cranial window in anesthetized rats, using laser-scanning two-dimension confocal microscopy. Video images of capillaries, arterioles, and venules were digitized off-line to measure microvessel diameter and labeled erythrocyte flow and velocity in parenchymal capillaries up to 200 microm beneath the brain surface. The method was used to analyze the rapid adaptation of microcirculation to a brief decrease in perfusion pressure. Twenty-second periods of forebrain ischemia were induced using the tour-vessel occlusion model in eight rats. EEG, arterial blood pressure, and body temperature were continuously controlled. In all conditions, labeled erythrocyte flow and velocity were both very heterogeneous in capillaries. During ischemia, capillary perfusion was close to 0, but a low blood flow persisted in arterioles and venules, while EEG was flattening. The arteriole and venule diameter did not significantly change. At the unclamping of carotid arteries, there was an instantaneous increase (by about 150%) of arteriole diameter. Capillary erythrocyte flow and velocity increased within 5 seconds, up to, respectively, 346 +/- 229% and 233 +/- 156% of their basal value. No capillary recruitment of erythrocytes was detected. All variables returned to their basal levels within less than 100 seconds after declamping. The data are discussed in terms of a possible involvement of shear stress in the reperfusion period.

Animals↗
PubMed2003

Dynamic deformation of migratory efferent lymph-derived cells "trapped" in the inflammatory microcirculation.

The cellular immune response depends on the delivery of lymphocytes from the lymph node to the peripheral site of antigenic challenge. During their passage through the inflammatory microcirculaton, the migratory cells can become transiently immobilized or "trapped" in small caliber vessels. In this report, we used intravital microscopy and temporal area mapping to define the dynamic deformation of efferent lymph-derived mononuclear cells trapped in the systemic inflammatory microcirculation. Mononuclear cells obtained from the efferent lymph draining the oxazolone-stimulated microcirculation were labeled with fluorescent dye and reinjected into the feeding arterial circulation. Intravital video microscopy observed thousands of cells passing through the microcirculation; 35 cells were "trapped" in the oxazolone-stimulated microcirculation. Temporal area maps of the trapped cells demonstrated dramatic slowing and deformation. The cells were trapped in the microcirculation for a median of 8.90 sec (range 5-17 sec) prior to returning to the flow stream. During this period, the cells showed sustained movement associated with both antegrade locomotion (mean cell velocity = 7.92 microm/sec; range 1.16-14.23 microm/sec) and dynamic elongation (median cell length = 73.8 microm; range 58-144 microm). In contrast, efferent lymph-derived cells passing unimpeded through the microcirculation demonstrated rapid velocity (median velocity = 216 microm/sec) and spherical geometry (median diameter = 14.6 microm). Further, the membrane surface area of the "trapped" cells, calculated based on digital image morphometry and corrosion cast scanning electron microscopy, suggested that the fractional excess membrane of the cells in the efferent lymph was significantly greater than previous estimates of membrane excess. These data indicate that transient immobilization of efferent lymph-derived mononuclear cells in the systemic inflammatory microcirculation is rare. When "trapping" does occur, the shape changes and sustained cell movement facilitated by excess cell membrane may contribute to the return of the "trapped cells" into the flow stream.

Adjuvants, Immunologic↗
PubMed2001

Expression and immunolocalization of aquaporin water channels in rat exocrine pancreas.

Both the acinar and ductal cells of the pancreas secrete a near-isotonic fluid and may thus be sites of aquaporin (AQP) water channel expression. Northern blot analysis of mRNA from whole rat pancreas revealed high levels of AQP1 and AQP8 expression, whereas lower levels of AQP4 and AQP5 expression were just detectable by RT-PCR Southern blot analysis. Immunohistochemistry showed that AQP1 is localized in the microvasculature, whereas AQP8 is confined to the apical pole of the acinar cells. No labeling of acinar, ductal, or vascular tissue was detected with antibodies to AQP2-7. With immunoelectron microscopy, AQP8 labeling was observed not only at the apical membrane of the acinar cells but also among small intracellular vesicles in the subapical cytoplasm, suggesting that there may be regulated trafficking of AQP8 to the apical plasma membrane. To evaluate the contribution of AQPs to the membrane water permeability, video microscopy was used to measure the swelling of acinar cells in response to hypotonic stress. Osmotic water permeability was reduced by 90% following exposure to Hg(2+). Since AQP8 is confined to the apical membrane, the marked effect of Hg(2+) suggests that other water channels may be expressed in the basolateral membrane.

Algorithms↗
PubMed1988

Target-induced changes in macrophage migration may explain differences in lytic sensitivity among simian virus 40-transformed fibroblasts.

Time-lapse video microscopy has been used to study macrophage movement in the presence of SV40-transformed fibroblasts. In all, five different SV40-transformed cell lines (an uncloned line and four clones derived from it) were tested for their affects on the movement of LPS-activated macrophages (AM). Conditions for video microscope recording were designed to simulate, as best as possible, the conditions used in a 51Cr-release cytotoxicity assay. Our analysis shows that these targets had a range of effects on macrophage migration, from stimulation to complete inhibition of movement. Two of the targets we tested (SV-COL and SV-COL-E8) both highly sensitive to lysis, stimulated macrophage movement, inducing an "agitated" response. Two other targets (SV-COL-F11 and SV-COL-H5), both of intermediate sensitivity had, for the most part, a suppressive effect on macrophage movement. Finally, we found that one target, clone SV-COL-F5, which is completely resistant to lysis by AM, was able to completely inhibit macrophage movement. This inhibitory effect was accompanied by a "pathologic" change in the structure of the macrophage cytoplasm suggesting that this target escapes lysis in vitro by secreting a factor that incapacitates the effector cell. Overall, these observations suggest that target cell products that can affect the behavior of the AM may be important in determining the lytic sensitivity of transformed target cells.

Animals↗
PubMed2004

Ultrastructure in cell biology: do we still need it?

John Heuser is being honored in this special issue for his enormous contributions to cell biology using morphological approaches. Foremost in this context is his ability to use light and electron microscopy to visualize structures and processes such that the information has both scientific and artistic value. The beauty of his images helps to focus the observer more intensely on the scientific messages, which have been numerous and important. His recent studies of living cells using state-of-the-art light and video microscopy fits into a general pattern of a huge explosion in the application of these methods worldwide that is revolutionizing cell biology. However, whereas John Heuser continues to use light microscopy (LM) for a low-resolution global and dynamical overview he then moves on to the electron microscopy (EM) level to see the details; in this he is--unfortunately--in a minority; and EM is an approach that a majority of today's cell biologists never use. The continued drop in EM usage has already been articulated in recent reviews. Here, I suggest that an additional problem for EM in cell biology, in its continued crises, is the declining number of scientists who can confidently interpret the--admittedly--complex information in most electron micrographs of cells. A major re-education is needed, or cell biology as a discipline will have a real problem in the 21st century.

Animals↗
PubMed2001

Role of platelet glycoprotein IIb/IIIa in ADP-activated platelet adhesion to aortic endothelial cells in vitro: observation with video-enhanced contrast microscopy.

Intravascular thrombus formation downstream of cerebral arterial occlusion may result in necrosis of ischemic tissue. To clarify the causative mechanisms, interaction between adenosine-5'-diphosphate (ADP)-activated platelets and cultured human aortic endothelial cells (HAEC) was examined by employing video enhanced contrast-differential interference contrast (VEC-DIC) microscopy. The numbers of (1) control/platelets, (2) ADP-activated platelets, (3) ADP-activated, anti-platelet GP Ibalpha antibody (GUR20-5)-treated platelets, and (4) ADP-activated, platelet GP IIb/IIa antagonist (TAK-029)-treated platelets, associated with HAEC after superfusion and wash-out were counted in visual fields of 30 x 30 microm2. Many ADP-activated platelets adhered to HAEC directly, while almost no platelets adhered to HAEC in the control. The adhesion was almost completely blocked by the GP IIb/IIIa antagonist, but not by GP Ibalpha antibody. We conclude that initial binding of ADP-activated platelets to HAEC is mediated by platelet GP IIb/IIIa in this in vitro system.

Adenosine Diphosphate↗
PubMed1990

Kinetic analysis of mitotic spindle elongation in vitro.

Studies of mitotic spindle elongation in vitro using populations of diatom spindles visualized with immunofluorescence microscopy have shown that the two interdigitating half-spindles are driven apart by an ATP-dependent process that generates force in the zone of overlap between half-spindles. To characterize further the system responsible for spindle elongation, we observed spindle elongation directly with polarized light or phase-contrast video-microscopy. We report that the kinetics of spindle elongation versus time are linear. A constant rate of spindle elongation occurs despite the continuous decrease in length of the zone of overlap between half-spindles. The average rate of spindle elongation varies as a function of treatment, and rates measured match spindle elongation rates measured in vivo. When spindles elongated in the presence of polymerizing tubulin (from bovine brain), the extent of elongation was greater than the original zone of half-spindle overlap, but the rate of elongation was constant. No component of force due to tubulin polymerization was found. The total elongation observed in the presence of added tubulin could exceed a doubling of original spindle length, matching the elongation in the intact diatom. The linear rate of spindle elongation in vitro suggests that the force transducer for anaphase B is a mechanochemical ATPase, analogous to dynein or myosin, and that the force for spindle elongation does not arise from stored energy, e.g. in an elastic matrix in the midzone. Additionally, on the basis of observations described here, we conclude that the force-transduction system for spindle elongation must be able to remain in the zone of microtubule overlap during the sliding apart of half-spindles, and that the transducer can generate force between microtubules that are not strictly antiparallel.

Adenosine Triphosphate↗
PubMed2001

Lysis of prostate carcinoma cells by trifunctional bispecific antibodies (alpha EpCAM x alpha CD3).

Bispecific monoclonal antibodies (bsAbs) are a promising immunotherapeutic option for treatment of cancer, especially in situations of minimal residual disease. The combination of an anti-CD3 and anti-tumor-associated antigen antibody redirects cytotoxic T-lymphocytes towards malignant cells. Using a trifunctional bispecific antibody against EpCAM x CD3, that additionally activates Fc gamma R(+) accessory cells via its Fc region, we investigated the interaction between three EpCAM(+) prostate carcinoma cell lines and peripheral blood mononuclear cells (PBMCs) of healthy donors and patients with prostate carcinoma (PC). Visualization was performed by double immunocytochemical methods and computerized sequential video microscopy. Tumor cells and PBMCs supplemented with alpha EpCAM x alpha CD3 in 16-well chamber slides resulted in lysis of tumor cells within 1--3 days without any differences between patient and healthy donor PBMCs. The characteristic necrotic way of tumor cell killing (rounding, swelling, disrupting) could be observed in computerized sequences of video frames. Simultaneously, we could not reveal any form of apoptotic signal using three different apoptotic markers (TUNEL, M30 cyto death, anti-active caspase 3). Within the first 48 hr we observed typical PBMC cluster formation with increasing cell proliferation. PBMCs surrounding the tumor cells were not dominated by CD4(+), CD8(+), or CD14(+) cells. Lymphocytes with pore-forming perforin proteins concentrated towards the tumor target cells. Our combination of double immunocytochemical and computerized video microscopic techniques may serve as an important improvement of validity of cell-cell interaction experiments using in vitro models. (J Histochem Cytochem 49:911-917, 2001)

Antibodies, Bispecific↗
PubMed2005

Mislocalization of DNAH5 and DNAH9 in respiratory cells from patients with primary ciliary dyskinesia.

RATIONALE: Primary ciliary dyskinesia (PCD) is a genetically heterogeneous disorder characterized by recurrent infections of the airways and situs inversus in half of the affected offspring. The most frequent genetic defects comprise recessive mutations of DNAH5 and DNAI1, which encode outer dynein arm (ODA) components. Diagnosis of PCD usually relies on electron microscopy, which is technically demanding and sometimes difficult to interpret. METHODS: Using specific antibodies, we determined the subcellular localization of the ODA heavy chains DNAH5 and DNAH9 in human respiratory epithelial and sperm cells of patients with PCD and control subjects by high-resolution immunofluorescence imaging. We also assessed cilia and sperm tail function by high-speed video microscopy. RESULTS: In normal ciliated airway epithelium, DNAH5 and DNAH9 show a specific regional distribution along the ciliary axoneme, indicating the existence of at least two distinct ODA types. DNAH5 was completely or only distally absent from the respiratory ciliary axoneme in patients with PCD with DNAH5- (n = 3) or DNAI1- (n = 1) mutations, respectively, and instead accumulated at the microtubule-organizing centers. In contrast to respiratory cilia, sperm tails from a patient with DNAH5 mutations had normal ODA heavy chain distribution, suggesting different modes of ODA generation in these cell types. Blinded investigation of a large cohort of patients with PCD and control subjects identified DNAH5 mislocalization in all patients diagnosed with ODA defects by electron microscopy (n = 16). Cilia with complete axonemal DNAH5 deficiency were immotile, whereas cilia with distal DNAH5 deficiency showed residual motility. CONCLUSIONS: Immunofluorescence staining can detect ODA defects, which will possibly aid PCD diagnosis.

Axonemal Dyneins↗
PubMed2000

Formation of three-dimensional thyroid follicle-like structures by polarized FRT cells made communication competent by transfection and stable expression of the connexin-32 gene.

Pig thyrocytes, either in the intact gland or cultured under conditions leading to thyroid follicle reconstitution, coexpress two gap junction proteins, connexin-32 (Cx32) and connexin-43 (Cx43). As thyrocytes cultured in the form of a monolayer only express Cx43, we hypothesized that Cx32 could play a role in thyroid folliculogenesis. In the present work, we analyzed the ability of polarized FRT cells (that are gap junction deficient) to form follicle-like structures after stable transfection with either Cx32 or Cx43 genes. Wild-type and transfected FRT cells, while growing, showed the capacity to form three-dimensional structures corresponding to domes that result from the accumulation of fluid underneath limited areas of the cell layer. The number of domes formed by FRT cells expressing Cx32 (FRT-Cx32) was 2- to 3-fold higher than that obtained with either wild-type or Cx43-transfected FRT cells (FRT-Cx43). Domes generated by FRT-Cx32 cells were stable (beyond 3 weeks of culture), whereas those formed from wild-type or FRT-Cx43 cells were transient, disappearing when cells reached confluence. Inspection of the cell organization within domes formed from FRT-Cx32 cells by phase contrast and confocal microscopy revealed a progressive transition from domes toward closed structures with a lumen. The tightness of the lumen was demonstrated by the retention of a fluorescent probe, lucifer yellow, introduced by microinjection. Electron microscope examinations showed that the neoformed follicle-like structures had an inside-out polarity. Analyses of cell motion and division with time, by fluorescence video microscopy, indicated that the transformation of domes into inside-out follicles brings into play the migration of cells and, to a lesser extent, cell multiplication underneath the domes. In conclusion, FRT cells forced to express Cx32 give rise to domes that transform into closed inside-out follicles. This gain of function appears Cx specific, as FRT-Cx43 cells did not form similar structures. Our data suggest that the formation and/or functioning of Cx32 gap junctions might represent a key event in thyroid epithelium morphogenesis, i.e. formation of a lumen from a tight epithelial cell layer.

Animals↗
PubMed1985

Transcapillary diffusion of Na-fluorescein measured by a 'large window technique' in skin areas of the forefoot.

A method is introduced for the study of transcapillary diffusion of 20% Na-fluorescein (0.3 ml/l of blood given intravenously) in skin areas by a fluorescence video-microscopy system which has been used earlier for measurements at the single capillary level or on axes crossing capillary groups. Two groups of 17 and 14 healthy volunteers respectively were included in a test-retest experiment (first measurement at day one, second measurement at day three). The appearance of the dye was visualized and the intensity of fluorescent light measured by a quadratic video densitometer window at the forefoot in an area of 2 mm2 containing 78 capillaries on the average. In both groups with and without control of the skin temperature, there were no statistically significant differences of the mean values measured at the first and second day of study. However, the mean intraindividual differences and the coefficients of variance were significantly (p less than 0.005) higher in the subjects without temperature control. It is concluded that the method is suited to detect transcapillary diffusion of Nafluorescein in skin areas by an almost atraumatic procedure with acceptable reproducibility. Potentially, it may be used to follow the natural history of microvascular diseases and to test the effect of various treatment modalities.

Adult↗
PubMed1993

Fucoidin, but not yeast polyphosphomannan PPME, inhibits leukocyte rolling in venules of the rat mesentery.

Leukocyte rolling in venules is inhibited by several sulfated polysaccharides, by antibodies to the leukocyte adhesion receptor L-selectin (LECAM-1), and by recombinant soluble L-selectin. The sulfated fucose polymer fucoidin and the polyphosphomannan PPME bind to L-selectin and inhibit L-selectin-mediated lymphocyte adhesion to lymph node high endothelial venules (LN-HEV). We investigated whether fucoidin and PPME also inhibit leukocyte rolling. Rolling leukocyte flux was determined by intravital microscopy in 47 venules (diameter 21 to 50 microns) of the rat mesentery with and without micro-infusion of each reagent through 8-microns glass micropipettes. Micro-infusion (1 mg/mL) or intravenous (IV) injection (25 mg/kg) of fucoidin, but not vehicle, reduced leukocyte rolling by greater than 90%. The half-effective concentration was approximately 2.5 micrograms/mL. Stroboscopic fluorescence video microscopy showed that fucoidin decreased the fraction of rolling leukocytes from 44% of all leukocytes passing the venules in control to less than 1%. PPME micro-infusion (1 mg/mL) or IV injection (14 mg/kg) did not reduce leukocyte rolling. Hence, leukocyte rolling differs from lymphocyte homing with respect to the effect of PPME. This may be related to fucoidin binding to L-selectin with greater affinity than PPME. Alternatively, inflamed venular endothelium may express a ligand for L-selectin different from that constitutively expressed on LN-HEV.

Animals↗
PubMed1997

Comparable effects of arteriolar and capillary stimuli on blood flow in rat skeletal muscle.

Although the capillary wall represents an active interface between blood and tissue, the potential role of the capillary in blood flow control has not been determined. The goals were (i) to establish the presence of the capillary sensing and communication phenomenon (Dietrich and Tyml, Microvasc. Res. 43, 87-99, 1992) in mammalian microvasculature and (ii) to determine the relative sensitivity of the capillary and the arteriole to locally applied vasoactive agents. Using intravital video microscopy, norepinephrine (NE; 10(-7)-3 x 10(-3) M), acetylcholine (ACh; 10(-4)-10(-2) M), or bradykinin (BK; 10(-9)-10(-3) M) was applied via micropipettes on capillaries (300 microm downstream from feeding arterioles) or on arterioles, at the surface of the extensor digitorum longus muscle of anesthetized rats. Red blood cell velocity (VRBC) in capillaries and arteriolar diameters was measured from video recordings. The overall control VRBC and control diameter were 190 microm/sec and 8.3 microm, respectively. NE applied on the capillary caused a dose-dependent reduction in VRBC (up to 100%, i.e., 0 microm/sec) via a constriction of the feeding arteriole. Both ACh and BK applied on the capillary caused a dose-dependent increase in VRBC (up to 115%) via arteriolar dilation. Based on two different approaches, these responses could not be explained in terms of diffusion of agents from capillary to the arteriole. When testing for the relative sensitivity of the arteriole and the capillary, application of NE and ACh on arterioles caused VRBC and diameter responses similar to those of capillary stimulations. When testing for the speed of response in these two microvessels, the time of noticeable VRBC change after NE (i.e., 10% from control) was also similar. We concluded that (i) the rat skeletal muscle capillary could respond to a variety of locally applied materials and (ii) the capillary could have as profound an effect on microvascular flow as the arteriole. Thus capillary could have the potential to participate in microvascular flow control.

Acetylcholine↗
PubMed1990

Keratin filaments restrict organelle migration into the forming spindle of newt pneumocytes.

When viewed by light microscopy the mitotic spindle of newt pneumocytes appears to assemble in an optically clear area of cytoplasm, virtually devoid of mitochondria and other organelles, which is often much larger than the spindle. This clear area is also frequently larger than the region previously occupied by the nucleus. It forms even in prometaphase cells depleted of microtubules prior to nuclear envelope breakdown by colchicine treatment. Time-lapse video microscopy reveals that as prometaphase proceeds this clear area slowly and progressively collapses around the forming spindle so that it is greatly diminished or nonexistent by the onset of anaphase. The sharply defined nature of the boundary between the clear area and the remaining cytoplasm and the fact that organelles accumulate at its periphery suggest that a structural barrier is present at the boundary that limits organelle migration into the forming spindle. Immunofluorescence and electron microscopy, of cells previously followed in the living state, reveal that the periphery of the clear area contains prominent bundles of keratin filaments but lacks microtubules and actin. From our observations we conclude that keratin filaments form a loosely organized cage that surrounds the forming newt pneumocyte spindle. We propose that this cage functions, in part, to restrict the dispersion of chromosomes during nuclear envelope breakdown and to impede the bulk migration of organelles into the forming spindle.

Animals↗
PubMed1996

Actin-based motility of isolated axoplasmic organelles.

We previously showed that axoplasmic organelles from the squid giant axon move toward the barbed ends of actin filaments and that KI-washed organelles separated from soluble proteins by sucrose density fractionation retain a 235-kDa putative myosin. Here, we examine the myosin-like activities of KI-washed organelles after sucrose density fractionation to address the question whether the myosin on these organelles is functional. By electron microscopy KI-washed organelles bound to actin filaments in the absence of ATP but not in its presence. Analysis of organelle-dependent ATPase activity over time and with varying amounts of organelles revealed a basal activity of 350 (range: 315-384) nmoles Pi/mg/min and an actin-activated activity of 774 (range: 560-988) nmoles/mg/min, a higher specific activity than for the other fractions. By video microscopy washed organelles moved in only one direction on actin filaments with a net velocity of 1.11 +/- .03 microns/s and an instantaneous velocity of 1.63 +/- 0.29 microns/s. By immunogold electronmicroscopy, 7% of KI-washed organelles were decorated with an anti-myosin antibody as compared to 0.5% with non-immune serum. Thus, some axoplasmic organelles have a tightly associated myosin-like activity.

Actins↗
PubMed1988

Time-lapse photography: a novel tool to study normal morphogenesis and the diseased hepatobiliary system in children.

The etiology of biliary atresia (BA) and many other childhood cholangiopathies remains unknown. To investigate the normal and diseased hepatobiliary system in vitro, we isolated and cultured biliary epithelial cells (BEC) from a number of species including man. Time-lapse video microscopy was used to study cell movements in vitro, and demonstrated the characteristic morphologic appearance of the BEC and their interactions with hepatocytes and other cell types on artificial biosynthetic materials. The cells remain attached and viable in routine culture conditions; however, when placed onto or within a source of extracellular matrix, the cells move off the surface and form characteristic morphologic patterns, which on time-lapse video are consistent with tube or duct formation. These findings suggest that time-lapse photography may be a useful tool for in vitro studies of normal morphogenesis of the hepatobiliary system and may enable us to better understand biliary atresia and other childhood cholangiopathies.

Animals↗