Project description:To examine global gene expression profile of chicken early paraxial mesoderm differentiation, we microdissected stage 12HH chicken PSM regions into 20 pieces (10 pieces both left-right PSM), including the tail bud, the PSM and somites. We create microarray series using these fragments.
Project description:To examine global gene expression profile of chicken early paraxial mesoderm differentiation, we microdissected stage 12HH chicken PSM regions into 20 pieces (10 pieces both left-right PSM), including the tail bud, the PSM and somites. We create microarray series using these fragments. Duplicated 10 fragmented tissues from stage 12 chicken PSM regions. contributor: IGBMC microarray facility
Project description:Stem cell-derived tissues have wide potential for modelling developmental and pathological processes as well as cell-based therapy. However, it has proven difficult to generate several key cell types in vitro, including skeletal muscle. In vertebrates, skeletal muscles derive during embryogenesis from the presomitic mesoderm (PSM). Using PSM development as a guide to establish conditions for the differentiation of monolayer cultures of embryonic stem (ES) cells into PSM-like cells without the introduction of transgenes or cell sorting. We generated a high resolution transcriptome expression landscape along the PSM of the mouse embryo, by microdissecting consecutive fragment of the PSM along the antero-posterior axis of the embryo. We took advantage of the observation that during development of embryo, the antero-posterior spatial position of the tissue is directly correlated to its differentiation (time) stage, thus generating an expression time-course of presomitic mesoderm development.
Project description:Stem cell-derived tissues have wide potential for modelling developmental and pathological processes as well as cell-based therapy. However, it has proven difficult to generate several key cell types in vitro, including skeletal muscle. In vertebrates, skeletal muscles derive during embryogenesis from the presomitic mesoderm (PSM). Treatment of mouse ES cells with a combination of the secreted Wnt activator R-Spondin3 and the BMP inhibitor Noggin generated cells expressing the early PSM marker Mesogenin1 (Msgn1) with high efficiency. To confirm their identity, we mapped gene expression profiles at successive stages of PSM differentiation in vivo and showed that the differentiated ES cells closely corresponded to the posterior PSM domain that expresses Msgn1 and then with time in culture matured to acquire the profile of the anterior Pax3 domain. When grafted into injured adult muscle in vivo these Pax3-expressing-cells generated large numbers of muscle fibers. Our system therefore efficiently produces myogenic precursors in vitro by recapitulating stepwise early myogenic differentiation in vivo. These findings should advance the development of cellular therapies for muscle degenerative diseases. Pre Somitic Mesoderm (PSM) were dissected into 7 pieces in Stage E8.5 Mouse embryo. The experiment was designed to have biological triplicate in each pieces. The PSM is a symmetric structure, left and right area were obtained from one embryo, another set of dissection was obtained from another embryo.
Project description:4 microarray time series was generated to identify cyclic genes of the segmentation clock in the mouse (2 time series), the chicken and the zebrafish. The right posterior half presomitic mesoderms (PSM) from 20 mouse embryos, 18 chicken embryos and 21 zebrafish embryos were dissected while the contralateral side of the embryo containing the left PSM was immediately fixed to be analyzed by in situ hybridization using a Lfng (fot mosue and chicken) or hes7 (zebrafish) probe to order the samples along the segmentation clock oscillation cycle. Probes were produced from RNA extracted from each of the dissected posterior half PSMs using a two-step amplification protocol and were hybridized to Affymetrix GeneChip MOE430A, MOE430 2.0, Affymetrix GeneChip chicken genome array, or Affymetrix GeneChip zebrafish array.