Parapseudoflavitalea muciniphila gen. nov., sp. nov., a member of the family Chitinophagaceae isolated from a human peritoneal tumour and reclassification of Pseudobacter ginsenosidimutans as Pseudoflavitalea ginsenosidimutans comb. nov.

A Gram-stain-negative, microaerophilic, non-motile, rod-shaped bacterium strain designated PMP191FT, was isolated from a human peritoneal tumour. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the organism formed a lineage within the family Chitinophagaceae that was distinct from members of the genus Pseudoflavitalea (95.1-95.2 % sequence similarity) and Pseudobacter ginsenosidimutans (94.4 % sequence similarity). The average nucleotide identity values between strain PMP191FT and Pseudoflavitalea rhizosphaerae T16R-265T and Pseudobacter ginsenosidimutans Gsoil 221T was 68.9 and 62.3% respectively. The only respiratory quinone of strain PMP191FT was MK-7 and the major fatty acids were iso-C15 : 0, iso-C15 : 1 G and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c). The polar lipids consisted of phosphatidylethanolamine and some unidentified amino and glycolipids. The G+C content of strain PMP191FT calculated from the genome sequence was 43.4 mol%. Based on phylogenetic, phenotypic and chemotaxonomic evidence, strain PMP191FT represents a novel species and genus for which the name Parapseudoflavitalea muciniphila gen. nov., sp. nov. is proposed. The type strain is PMP191FT (=DSM 104999T=ATCC BAA-2857T = CCUG 72691T). The phylogenetic analyses also revealed that Pseudobacter ginsenosidimutans shared over 98 % sequence similarly to members of the genus Pseudoflavitalea. However, the average nucleotide identity value between Pseudoflavitalea rhizosphaerae T16R-265T, the type species of the genus and Pseudobacter ginsenosidimutans Gsoil 221T was 86.8 %. Therefore, we also propose that Pseudobacter ginsenosidimutans be reclassified as Pseudoflavitalea ginsenosidimutans comb. nov.

Quantifying CD4 receptor protein in two human CD4+ lymphocyte preparations for quantitative flow cytometry.

BACKGROUND: In our previous study that characterized different human CD4+ lymphocyte preparations, it was found that both commercially available cryopreserved peripheral blood mononuclear cells (PBMC) and a commercially available lyophilized PBMC (Cyto-Trol™) preparation fulfilled a set of criteria for serving as biological calibrators for quantitative flow cytometry. However, the biomarker CD4 protein expression level measured for T helper cells from Cyto-Trol was about 16% lower than those for cryopreserved PBMC and fresh whole blood using flow cytometry and mass cytometry. A primary reason was hypothesized to be due to steric interference in anti- CD4 antibody binding to the smaller sized lyophilized control cells. METHOD: Targeted multiple reaction monitoring (MRM) mass spectrometry (MS) is used to quantify the copy number of CD4 receptor protein per CD4+ lymphocyte. Scanning electron microscopy (SEM) is utilized to assist searching the underlying reasons for the observed difference in CD4 receptor copy number per cell determined by MRM MS and CD4 expression measured previously by flow cytometry. RESULTS: The copy number of CD4 receptor proteins on the surface of the CD4+ lymphocyte in cryopreserved PBMCs and in lyophilized control cells is determined to be (1.45 ± 0.09) × 10(5) and (0.85 ± 0.11) × 10(5), respectively, averaged over four signature peptides using MRM MS. In comparison with cryopreserved PBMCs, there are more variations in the CD4 copy number in lyophilized control cells determined based on each signature peptide. SEM images of CD4+ lymphocytes from lyophilized control cells are very different when compared to the CD4+ T cells from whole blood and cryopreserved PBMC. CONCLUSION: Because of the lyophilization process applied to Cyto-Trol control cells, a lower CD4 density value, defined as the copy number of CD4 receptors per CD4+ lymphocyte, averaged over three different production lots is most likely explained by the loss of the CD4 receptors on damaged and/or broken microvilli where CD4 receptors reside. Steric hindrance of antibody binding and the association of CD4 receptors with other biomolecules likely contribute significantly to the nearly 50% lower CD4 receptor density value for cryopreserved PBMC determined from flow cytometry compared to the value obtained from MRM MS.

On the linear stability of blood flow through model capillary networks.

Under the approximation that blood behaves as a continuum, a numerical implementation is presented to analyze the linear stability of capillary blood flow through model tree and honeycomb networks that are based on the microvascular structures of biological tissues. The tree network is comprised of a cascade of diverging bifurcations, in which a parent vessel bifurcates into two descendent vessels, while the honeycomb network also contains converging bifurcations, in which two parent vessels merge into one descendent vessel. At diverging bifurcations, a cell partitioning law is required to account for the nonuniform distribution of red blood cells as a function of the flow rate of blood into each descendent vessel. A linearization of the governing equations produces a system of delay differential equations involving the discharge hematocrit entering each network vessel and leads to a nonlinear eigenvalue problem. All eigenvalues in a specified region of the complex plane are captured using a transformation based on contour integrals to construct a linear eigenvalue problem with identical eigenvalues, which are then determined using a standard QR algorithm. The predicted value of the dimensionless exponent in the cell partitioning law at the instability threshold corresponds to a supercritical Hopf bifurcation in numerical simulations of the equations governing unsteady blood flow. Excellent agreement is found between the predictions of the linear stability analysis and nonlinear simulations. The relaxation of the assumption of plug flow made in previous stability analyses typically has a small, quantitative effect on the stability results that depends on the specific network structure. This implementation of the stability analysis can be applied to large networks with arbitrary structure provided only that the connectivity among the network segments is known.

Desorption electrospray ionization imaging mass spectrometry as a tool for investigating model prebiotic reactions on mineral surfaces.

Mineral-assisted thermal decomposition of formamide (HCONH(2)) is a heavily studied model prebiotic reaction that has offered valuable insights into the plausible pathways leading to the chemical building blocks of primordial informational polymers. To date, most efforts have focused on the analysis of formamide reaction products released in solution, although several studies have examined the role of mineral catalysts in promoting this chemistry. We show here that the direct investigation of reactive mineral surfaces by desorption electrospray ionization-mass spectrometry imaging (DESI-MSI) gives a new perspective on the important role of the mineral surface in the formation of reaction products. As a proof-of-principle example, we show that DESI-MSI allows interrogation of the molecular products produced on heterogeneous granite samples with minimal sample preparation. Purine and pyrimidine nucleobases and their derivatives are successfully detected by DESI-MSI, with a strong correlation of the spatial product distribution with the mineral microenvironment. To our knowledge, this study is the first application of DESI-MSI to the study of complex and porous mineral surfaces and their roles in chemical evolution. This DESI-MSI approach is generally applicable to a wide range of reactions or other processes involving minerals.

EDS measurements of X-ray intensity at WDS precision and accuracy using a silicon drift detector.

The accuracy and precision of X-ray intensity measurements with a silicon drift detector (SDD) are compared with the same measurements performed on a wavelength dispersive spectrometer (WDS) for a variety of elements in a variety of materials. In cases of major (>0.10 mass fraction) and minor (>0.01 mass fraction) elements, the SDD is demonstrated to perform as well or better than the WDS. This is demonstrated both for simple cases in which the spectral peaks do not interfere (SRM-481, SRM-482, and SRM-479a), and for more difficult cases in which the spectral peaks have significant interferences (the Ba L/Ti K lines in a series of Ba/Ti glasses and minerals). We demonstrate that even in the case of significant interference high count SDD spectra are capable of accurately measuring Ti in glasses with Ba:Ti mass fraction ratios from 2.7:1 to 23.8:1. The results suggest that for many measurements wavelength spectrometry can be replaced with an SDD with improved accuracy and precision.

An examination of kernite (Na2B4O6(OH)2·3H2O) using X-ray and electron spectroscopies: quantitative microanalysis of a hydrated low-Z mineral.

Mineral borates, the primary industrial source of boron, are found in a large variety of compositions. One such source, kernite (Na2B4O6(OH)2·3H2O), offers an array of challenges for traditional electron-probe microanalysis (EPMA)-it is hygroscopic, an electrical insulator, composed entirely of light elements, and sensitive to both low pressures and the electron beam. However, the approximate stoichiometric composition of kernite can be analyzed with careful preparation, proper selection of reference materials, and attention to the details of quantification procedures, including correction for the time dependency of the sodium X-ray signal. Moreover, a reasonable estimation of the mineral's water content can also be made by comparing the measured oxygen to the calculated stoichiometric oxygen content. X-ray diffraction, variable-pressure electron imaging, and visual inspection elucidate the structural consequences of high vacuum treatment of kernite, while Auger electron spectroscopy and X-ray photoelectron spectroscopy confirm electron beam-driven migration of sodium and oxygen out of the near-surface region (sampling depth ≈ 2 nm). These surface effects are insufficiently large to significantly affect the EPMA results (sampling depth ≈ 400 nm at 5 keV).

Bridging the micro-to-macro gap: a new application for micro X-ray fluorescence.

X-ray elemental mapping and X-ray spectrum imaging are powerful microanalytical tools. However, their scope is often limited spatially by the raster area of a scanning electron microscope or microprobe. Limited sampling size becomes a significant issue when large area (>10 cm²), heterogeneous materials such as concrete samples or others must be examined. In such specimens, macro-scale structures, inclusions, and concentration gradients are often of interest, yet microbeam methods are insufficient or at least inefficient for analyzing them. Such requirements largely exclude the samples of interest presented in this article from electron probe microanalysis. Micro X-ray fluorescence-X-ray spectrum imaging (µXRF-XSI) provides a solution to the problem of macro-scale X-ray imaging through an X-ray excitation source, which can be used to analyze a variety of large specimens without many of the limitations found in electron-excitation sources. Using a mid-sized beam coupled with an X-ray excitation source has a number of advantages, such as the ability to work at atmospheric pressure and lower limits of detection owing to the absence of electron-induced bremsstrahlung. µXRF-XSI also acts as a complement, where applicable, to electron microbeam X-ray output, highlighting areas of interest for follow-up microanalysis at a finer length scale.

Fluorinated copolymer nanoparticles for multimodal imaging applications.

Nanomaterials have emerged as valuable tools in biomedical imaging techniques. Here, the synthesis and characterization of a novel fluorinated nanoparticle with potential applications as an MRI contrast agent is reported. Particles were synthesized using a free radical polymerization technique. Secondary ion mass spectrometry analysis showed that the particles' surface contained fluorinated groups and nitrogen-containing groups. Solid-state NMR spectroscopy suggested the presence of two distinct fluorine resonances, which conforms to the structure of the fluorinated monomer. Ongoing studies aim to evaluate the performance of the nanoparticles as MRI contrast agents both in vitro and in vivo.

Functional Tissue Engineering: A Prevascularized Cardiac Muscle Construct for Validating Human Mesenchymal Stem Cells Engraftment Potential In Vitro.

The influence of somatic stem cells in the stimulation of mammalian cardiac muscle regeneration is still in its early stages, and so far, it has been difficult to determine the efficacy of the procedures that have been employed. The outstanding question remains whether stem cells derived from the bone marrow or some other location within or outside of the heart can populate a region of myocardial damage and transform into tissue-specific differentiated progenies, and also exhibit functional synchronization. Consequently, this necessitates the development of an appropriate in vitro three-dimensional (3D) model of cardiomyogenesis and prompts the development of a 3D cardiac muscle construct for tissue engineering purposes, especially using the somatic stem cell, human mesenchymal stem cells (hMSCs). To this end, we have created an in vitro 3D functional prevascularized cardiac muscle construct using embryonic cardiac myocytes (eCMs) and hMSCs. First, to generate the prevascularized scaffold, human cardiac microvascular endothelial cells (hCMVECs) and hMSCs were cocultured onto a 3D collagen cell carrier (CCC) for 7 days under vasculogenic culture conditions; hCMVECs/hMSCs underwent maturation, differentiation, and morphogenesis characteristic of microvessels, and formed dense vascular networks. Next, the eCMs and hMSCs were cocultured onto this generated prevascularized CCCs for further 7 or 14 days in myogenic culture conditions. Finally, the vascular and cardiac phenotypic inductions were characterized at the morphological, immunological, biochemical, molecular, and functional levels. Expression and functional analyses of the differentiated progenies revealed neo-cardiomyogenesis and neo-vasculogenesis. In this milieu, for instance, not only were hMSCs able to couple electromechanically with developing eCMs but were also able to contribute to the developing vasculature as mural cells, respectively. Hence, our unique 3D coculture system provides us a reproducible and quintessential in vitro 3D model of cardiomyogenesis and a functioning prevascularized 3D cardiac graft that can be utilized for personalized medicine.

Stability and transient dynamics of thin liquid films flowing over locally heated surfaces.

The dynamics and linear stability of a liquid film flowing over a locally heated surface are studied using a long-wave lubrication analysis. The temperature gradient at the leading edge of the heater induces a gradient in surface tension that opposes the gravitationally driven flow and leads to the formation of a pronounced capillary ridge. The resulting free-surface shapes are computed, and their stability to spanwise perturbations is analyzed for a range of Marangoni numbers, substrate inclination angles, and temperature profiles. Instability is predicted above a critical Marangoni number for a finite band of wave numbers separated from zero, which is consistent with published results from experiment and direct numerical simulation. An energy analysis is used to gain insight into the effect of inclination angle on the instability. Because the spatial nonuniformity of the base state gives rise to nonnormal linearized operators that govern the evolution of perturbations, a nonmodal, transient analysis is used to determine the maximum amplification of small perturbations to the film. The structure of optimal perturbations of different wave numbers is computed to elucidate the regions of the film that are most sensitive to perturbations, which provides insight into ways to stabilize the flow. The results of this analysis are contrasted to those for noninertial coating flows over substrates with topographical features, which exhibit similar capillary ridges but are strongly stable to perturbations.

Interaction of micrometer-scale particles with nanotextured surfaces in shear flow.

Dynamic particle adhesion from flow over collecting surfaces with nanoscale heterogeneity occurs in important natural systems and current technologies. Accurate modeling and prediction of the dynamics of particles interacting with such surfaces will facilitate their use in applications for sensing, separating, and sorting colloidal-scale objects. In this paper, the interaction of micrometer-scale particles with electrostatically heterogeneous surfaces is analyzed. The deposited polymeric patches that provide the charge heterogeneity in experiments are modeled as 11-nm disks randomly distributed on a planar surface. A novel technique based on surface discretization is introduced to facilitate computation of the colloidal interactions between a particle and the heterogeneous surface based on expressions for parallel plates. Combining these interactions with hydrodynamic forces and torques on a particle in a low Reynolds number shear flow allows particle dynamics to be computed for varying net surface coverage. Spatial fluctuations in the local surface density of the deposited patches are shown responsible for the dynamic adhesion phenomena observed experimentally, including particle capture on a net-repulsive surface.

A Novel Human Tissue-Engineered 3-D Functional Vascularized Cardiac Muscle Construct.

Organ tissue engineering, including cardiovascular tissues, has been an area of intense investigation. The major challenge to these approaches has been the inability to vascularize and perfuse the in vitro engineered tissue constructs. Attempts to provide oxygen and nutrients to the cells contained in the biomaterial constructs have had varying degrees of success. The aim of this current study is to develop a three-dimensional (3-D) model of vascularized cardiac tissue to examine the concurrent temporal and spatial regulation of cardiomyogenesis in the context of postnatal de novo vasculogenesis during stem cell cardiac regeneration. In order to achieve the above aim, we have developed an in vitro 3-D functional vascularized cardiac muscle construct using human induced pluripotent stem cell-derived embryonic cardiac myocytes (hiPSC-ECMs) and human mesenchymal stem cells (hMSCs). First, to generate the prevascularized scaffold, human cardiac microvascular endothelial cells (hCMVECs) and hMSCs were co-cultured onto a 3-D collagen cell carrier (CCC) for 7 days under vasculogenic culture conditions. In this milieu, hCMVECs/hMSCs underwent maturation, differentiation, and morphogenesis characteristic of microvessels, and formed extensive plexuses of vascular networks. Next, the hiPSC-ECMs and hMSCs were co-cultured onto this generated prevascularized CCCs for further 7 or 14 days in myogenic culture conditions. Finally, the vascular and cardiac phenotypic inductions were analyzed at the morphological, immunological, biochemical, molecular, and functional levels. Expression and functional analyses of the differentiated cells revealed neo-angiogenesis and neo-cardiomyogenesis. Thus, our unique 3-D co-culture system provided us the apt in vitro functional vascularized 3-D cardiac patch that can be utilized for cellular cardiomyoplasty.

Measurement and Modeling of the Ability of Crack Fillers to Prevent Chloride Ingress into Mortar.

A common repair procedures applied to damaged concrete is to fill cracks with an organic polymer. This operation is performed to increase the service life of the concrete by removing a preferential pathway for the ingress of water, chlorides, and other deleterious species. To effectively fulfill its mission of preventing chloride ingress, the polymer must not only fully fill the macro-crack, but must also intrude the damage zone surrounding the crack perimeter. Here, the performance of two commonly employed crack fillers, one epoxy, and one methacrylate, are investigated using a combined experimental and computer modeling approach. Neutron tomography and microbeam X-ray fluorescence spectrometry (µXRF) measurements are employed on pre-cracked and chloride-exposed specimens to quantify the crack filling and chloride ingress limiting abilities, respectively, of the two polymers. A two-dimensional model of chloride transport is derived from a mass balance and solved by the finite element method. Crack images provided by µXRF are used to generate the input microstructure for the simulations. When chloride binding and a time-dependent mortar diffusivity are both included in the computer model, good agreement with the experimental results is obtained. Both crack fillers significantly reduce chloride ingress during the 21 d period of the present experiments; however, the epoxy itself contains approximately 4 % by mass chlorine. Leaching studies were performed assess the epoxy as a source of deleterious ions for initiating corrosion of the steel reinforcement in concrete structures.

Statistically-based DLVO approach to the dynamic interaction of colloidal microparticles with topographically and chemically heterogeneous collectors.

Electrostatic surface heterogeneity on the order of a few nanometers is common in colloidal and bacterial systems, dominating adhesion and aggregation and inducing deviations from classical DLVO theory based on a uniform distribution of surface charge. Topographical heterogeneity and roughness also strongly influence adhesion. In this work, a model is introduced to quantify the spatial fluctuations in the interaction of microparticles in a flowing suspension with a wall aligned parallel to the flow. The wall contains nanoscale chemical and topographical heterogeneities ("patches") that are randomly distributed and produce localized attraction and repulsion. These attractive and repulsive regions induce fluctuations in the trajectories of the flowing particles that are critical to particle capture by the wall. The statistical distribution of patches is combined with mean-field DLVO calculations between a particle and two homogeneous surfaces: one with the surface potential of the patches and one with the potential of the underlying wall. These surface potentials could be obtained in experiments from zeta potential measurements for the bare wall and for one saturated with patches. This simple model reproduces the mean DLVO interaction force or energy vs. particle-wall separation distance, its variance, and particle adhesion thresholds from direct simulations of particle trajectories over patchy surfaces. The predictions of the model are consistent with experimental findings of significant microparticle deposition onto patchy, net-repulsive surfaces whose apparent zeta potential has the same sign as that of the particles. Deposition is significantly enhanced if the patches protrude even slightly from the surface. The model predictions are also in agreement with the observed variation of the adhesion threshold with the shear rate in published studies of dynamic microparticle adhesion on patchy surfaces.

A novel tubular scaffold for cardiovascular tissue engineering.

We have developed a counter rotating cone extrusion device to produce the next generation of three-dimensional collagen scaffold for tissue engineering. The device can produce a continuously varying fibril angle from the lumen to the outside of a 5-mm-diameter collagen tube, similar to the pattern of heart muscle cells in the intact heart. Our scaffold is a novel, oriented, type I collagen, tubular scaffold. We selected collagen because we believe there are important signals from the collagen both geometrically and biochemically that elicit the in vivo -like phenotypic response from the cardiomyocytes. We have shown that cardiomyocytes can be cultured in these tubes and resemble an in vivo phenotype. This new model system will provide important information leading to the design and construction of a functional, biologically based assist device.

Influence of attractive van der Waals interactions on the optimal excitations in thermocapillary-driven spreading.

Recent investigations of microfluidic flows have focused on manipulating the motion of very thin liquid films by modulating the surface tension through an applied streamwise temperature gradient. The extent to which the choice of contact line model affects the flow and stability of such thermocapillary-driven films is not completely understood. Regardless of the contact line model used, the linearized disturbance operator corresponding to the evolution of the film height is non-normal, and a generalized non-modal stability analysis is required. Surprisingly, early predictions of frontal instability that stemmed from conventional modal analysis of thermocapillary flow on a flat, infinite precursor film showed excellent agreement with experiment. Within the more rigorous framework provided by a generalized stability analysis, this work investigates the transient dynamics and amplification of optimal disturbances subject to a finite precursor film generated by attractive van der Waals forces. Convergence of the disturbance growth rates and perturbed shapes to the asymptotic solutions obtained by conventional linear stability analysis occurs early in the spreading process. In addition, the level of transient disturbance amplification is minimal. The equations governing thermocapillary-driven spreading exhibit a small degree of non-normality, which explains the source of agreement between modal theory and experiment. The more rigorous generalized stability analysis presented here, however, affords critical insight into the types of disturbances leading to maximum unstable growth and the exact influence of the contact line model used.

Influence of boundary slip on the optimal excitations in thermocapillary driven spreading.

Thin liquid films driven to spread on homogeneous surfaces by thermocapillarity can undergo frontal breakup and parallel rivulet formation with well-defined wavelength. Previous modal analyses have relieved the well-known divergence in stress that occurs at a moving contact line by matching the front region to a precursor film. Because the linearized disturbance operator is non-normal, a generalized, nonmodal analysis is required to probe film stability at all times. The effect of the contact line model on nonmodal stability has not been previously investigated. This work examines the influence of boundary slip on thermocapillary driven spreading using a transient stability analysis, which recovers the conventional modal results in the long-time limit. In combination with earlier work on thermocapillary driven spreading, this study verifies that the dynamics and stability of this system are rather insensitive to the choice of contact line model and that the leading eigenvalue is physically determinant, thereby assuring results that agree with the eigenspectrum. Modal results for the flat precursor film model are reproduced with appropriate choice of slip coefficient and contact line slope.

Stability of a volatile liquid film spreading along a heterogeneously-heated substrate.

The dynamics and stability of a thin, viscous film of volatile liquid flowing under the influence of gravity over a non-uniformly heated substrate are investigated using lubrication theory. Attention is focused on the regime in which evaporation balances the flow due to gravity. The film terminates above the heater at an apparent contact line, with a microscopically thin precursor film adsorbed due to the disjoining pressure. The film develops a weak thermocapillary ridge due to the Marangoni stress at the upstream edge of the heated region. As for spreading films, a more significant ridge is formed near the apparent contact line. For weak Marangoni effects, the film evolves to a steady profile. For stronger Marangoni effects, the film evolves to a time-periodic state. Results of a linear stability analysis reveal that the steady film is unstable to transverse perturbations above a critical value of the Marangoni parameter, leading to finger formation at the contact line. The streamwise extent of the fingers is limited by evaporation. The time-periodic profiles are always unstable, leading to the formation of periodically-oscillating fingers. For rectangular heaters, the film profiles after instability onset are consistent with images from published experimental studies.

DLVO interaction of colloidal particles with topographically and chemically heterogeneous surfaces.

The DLVO force and potential energy of interaction between microspheres and topographically and chemically heterogeneous surfaces in aqueous solution are computed using a modification of the surface element integration approach. The heterogeneous surface has an array of cylindrical pillars of varying height, diameter, and arrangement to model different nano-topographies. In agreement with previous studies, the nano-topography decreases the size of the potential energy barrier for unfavorable surfaces because the pillars limit the minimum separation distance. The influence of topography is significant even for pillars several nanometers high and is more pronounced if the surface potential of the pillar tops differs from that of the underlying surface. A new force- and energy-averaging model is introduced as a simple method to compute the mean interaction energy or force between the particle and a heterogeneous surface, which differs significantly from a mean-field approach based on the average or nominal surface potential. Small variations in topography are found to remove large energy barriers to colloidal deposition. These results help explain the increased attraction of patchy surfaces towards particles relative to expectations based on typical DLVO calculations, which is particularly significant for surfaces with adsorbed polyelectrolytes.

The mechanical coupling of adult marrow stromal stem cells during cardiac regeneration assessed in a 2-D co-culture model.

Postnatal cardiomyocytes undergo terminal differentiation and a restricted number of human cardiomyocytes retain the ability to divide and regenerate in response to ischemic injury. However, whether these neo-cardiomyocytes are derived from endogenous population of resident cardiac stem cells or from the exogenous double assurance population of resident bone marrow-derived stem cells that populate the damaged myocardium is unresolved and under intense investigation. The vital challenge is to ameliorate and/or regenerate the damaged myocardium. This can be achieved by stimulating proliferation of native quiescent cardiomyocytes and/or cardiac stem cell, or by recruiting exogenous autologous or allogeneic cells such as fetal or embryonic cardiomyocyte progenitors or bone marrow-derived stromal stem cells. The prerequisites are that these neo-cardiomyocytes must have the ability to integrate well within the native myocardium and must exhibit functional synchronization. Adult bone marrow stromal cells (BMSCs) have been shown to differentiate into cardiomyocyte-like cells both in vitro and in vivo. As a result, BMSCs may potentially play an essential role in cardiac repair and regeneration, but this concept requires further validation. In this report, we have provided compelling evidence that functioning cardiac tissue can be generated by the interaction of multipotent BMSCs with embryonic cardiac myocytes (ECMs) in two-dimensional (2-D) co-cultures. The differentiating BMSCs were induced to undergo cardiomyogenic differentiation pathway and were able to express unequivocal electromechanical coupling and functional synchronization with ECMs. Our 2-D co-culture system provides a useful in vitro model to elucidate various molecular mechanisms underpinning the integration and orderly maturation and differentiation of BMSCs into neo-cardiomyocytes during myocardial repair and regeneration.