Paracrine and epigenetic control of trophectoderm differentiation from human embryonic stem cells: the role of bone morphogenic protein 4 and histone deacetylases.

Our understanding of paracrine and epigenetic control of trophectoderm (TE) differentiation is limited by available models of preimplantation human development. Simple, defined media for selective TE differentiation of human embryonic stem cells (hESCs) were developed, enabling mechanistic studies of early placental development. Paracrine requirements of preimplantation human development were evaluated with hESCs by measuring lineage-specific transcription factor expression levels in single cells and morphological transformation in response to selected paracrine and epigenetic modulators. Bone morphogenic protein 4 (BMP4) addition to feeder-free pluripotent stem cells on matrigel frequently formed CDX2-positive TE. However, BMP4 or activin A inhibition alone also produced a mix of mesoderm and extraembryonic endoderm under these conditions. Further, BMP4 failed to form TE from adherent hESC maintained in standard feeder-dependent monolayers. Given that the efficiency and selectivity of BMP4-induced TE depended on medium components, we developed a basal medium containing insulin and heparin. In this medium, BMP4 induction of TE was dose dependent and with activin A inhibition by SB431542 (SB), approached 100% of cells. This paracrine stimulation of pluripotent cells transformed colony morphology from a cuboidal to squamous epithelium quantitatively on day 3, and produced significant multinucleated syncytiotrophoblasts by day 8. Addition of trichostatin A, a histone deacetylase (HDAC) inhibitor, reduced HDAC3, histone H3K9 methylation, and slowed differentiation in a dose-dependent manner. Modulators of BMP4- or HDAC-dependent signaling might adversely influence the timing and viability of early blastocyst developed in vitro. Since blastocyst development is synchronized to uterine receptivity, epigenetic regulators of TE differentiation might adversely affect implantation in vivo.

Spatio-temporal data fusion for 3D+T image reconstruction in cerebral angiography.

This paper provides a framework for generating high resolution time sequences of 3D images that show the dynamics of cerebral blood flow. These sequences have the potential to allow image feedback during medical procedures that facilitate the detection and observation of pathological abnormalities such as stenoses, aneurysms, and blood clots. The 3D time series is constructed by fusing a single static 3D model with two time sequences of 2D projections of the same imaged region. The fusion process utilizes a variational approach that constrains the volumes to have both smoothly varying regions separated by edges and sparse regions of nonzero support. The variational problem is solved using a modified version of the Gauss-Seidel algorithm that exploits the spatio-temporal structure of the angiography problem. The 3D time series results are visualized using time series of isosurfaces, synthetic X-rays from arbitrary perspectives or poses, and 3D surfaces that show arrival times of the contrasted blood front using color coding. The derived visualizations provide physicians with a previously unavailable wealth of information that can lead to safer procedures, including quicker localization of flow altering abnormalities such as blood clots, and lower procedural X-ray exposure. Quantitative SNR and other performance analysis of the algorithm on computational phantom data are also presented.