This 1999 edition of The Neural Crest contains comprehensive information about the neural crest, a structure unique to the vertebrate embryo, which has only a transient existence in early embryonic life. The ontogeny of the neural crest embodies the most important issues in developmental biology, as the neural crest is considered to have played a crucial role in evolution of the vertebrate phylum. Data that analyse neural crest ontogeny in murine and zebrafish embryos have been included in this revision. This revised edition also takes advantage of recent advances in our understanding of markers of neural crest cell subpopulations, and a full chapter is now devoted to cell lineage analysis. The major research breakthrough since the first edition has been the introduction of molecular biology to neural crest research, enabling an elucidation of many molecular mechanisms of neural crest development. This book is essential reading for students and researchers in developmental biology, cell biology, and neuroscience.

Offers readers an understanding of the development of neural crest cells, which is crucial as many birth defects and tumours are of neural crest origin. Delving into stem cells from different locations of the body, this book explores the best possible source of such cells for the use in medical applications.

This dissertation, "Quantitative Characterization of Mouse Embryonic Stem Cell State Transition" by Xibin, Lu, 盧希彬, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: It is known that on-and-off transcription factors (TFs) regulations play crucial roles in differentiation and reprogramming. Oct4, Sox2 and Nanog as well as their constituted gene networks cooperatively maintain ES cell pluripotency. Recently, TFs such as Nanog, Stella and Rex1 show heterogeneous expression in ES cell populations and such heterogeneity lead to different differentiation potential indicating that heterogeneous gene expression may control cell fate. Besides gene regulation triggered by TFs, intrinsic noise was also found to be a potential source in heterogeneity and stochastic cell fate determination. For safe clinical application of stem cell-based therapy, it is crucial to quantify these dynamic processes and transition rate during research and development. However, in most cases, the transition rate is too low to be measured and is often entangled with cell proliferation. In order to disentangle these processes, we established a Nanog-EGFP reporter ES cell line and quantified the transition rates between two subpopulations. We also developed a mathematical model involving cell state transition, proliferation and noise to facilitate deciphering this process. We first generated a dual reporter ES cell line (Nanog-EGFP and EF1α-H2B-mCherry) based on PiggyBac system. Using a simple ES cell differentiation process, differentiation and dedifferentiation could be obviously observed based on Nanog ES cell reporter. In order to quantify the transition and proliferation rate, we performed multiple batches of ES cell differentiation and dedifferentiation based with this reporter ES cell line. Mathematical models (drift-diffusion-growth equation) are also introduced to fit the data. The quantitative and nontrivial model predictions lead to new experiments and hypotheses. Combined the flow cytometry data with mathematical models, we are able to directly derive the most likely potential landscape (Waddington's landscape) based on Nanog distribution. Our results and approaches developed would provide more clues for future ES cell differentiation studies. In another project, we generated multiple ES cell reporters in which Nanog, Oct4 and Rex1 were all labeled with different fluorescence proteins. We differentiated the reporter ES cell line specifically into epiblast stem cell (EpiSC) based on previous protocols. Based on flow cytometry analysis, we found that, during the transition from ES cell to EpiSC state, all the three marker genes showed heterogeneous expression pattern, and three sub-populations can be identified at the EpiSC steady (later) state. The cell fate identity and potential converting ability into other identities like ES cell or inter-converting ability among three populations were being conducted based on chemical factors. In addition, we quantitatively characterized PiggyBac mediated multiplex gene transfer in mouse embryonic stem cell which lay a good foundation in above mentioned ES cell reporter generation. HS4 insulator was also characterized on the effect of protecting from promoter interference and gene silencing as well as transposition efficiency. Our results suggest that, even after the development of CRISPR and TALEN, transposon technique is still a powerful tool to study synthetic gene circuit in mammalian systems, especially for prototyping and complex circuits with many parts. DOI: 10.5353/th_b5351003 Subjects: Stem cells Ce

This volume comprehensively covers new technologies and methodologies that have appeared for the study of mouse development - Update of volume 225, Guide to Techniques in Mouse Development, edited by P.M. Wassarman and M.L. DePamphilis and published in 1993 - Comprehensively covers: - new techniques for the cryopreservation of gametes and embryos - production of transgenic and null (knockout) animals (use of ES cells) - generation of conditional/inducible mutant animals, use of gene-trap mutagenesis - analysis of allele-specific expression, use of new reporter constructs - humanizing of transgenic animals - transcript profiling of mouse development - imaging of mouse development - rederivation of animals and use of mouse genomics