Transcriptomic landscape of developing PSM from Tailbud to somite in E8.5 mouse embryo and ES
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: philippe moncuquet
PROVIDER: E-GEOD-39615 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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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.
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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. ES cellline E14 129P2/OlaHsd with DMSO, Rspo, and Noggin at Days 3 and 4
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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.
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Project description:A microarray time series was generated to identify cyclic genes of the segmentation clock in the mouse. The right posterior half presomitic mesoderms (PSM) from 17 mouse 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 probe to order the samples along the segmentation clock oscillation cycle. Probes were produced from RNA extracted from the 17 dissected posterior half PSMs using a two-step amplification protocol and were hybridized to Affymetrix GeneChip MOE430A. The reproducibility of the amplification procedure was initially assessed by comparing array data generated from the right and the left posterior PSM from the same embryo. Because of the symmetry of the paraxial mesoderm along the left-right axis, left and right samples are expected to show overtly similar gene expression. RNA was amplified from three such sample pairs (1, a and b; 2, a and b; 3, a and b) and hybridized on Murine Genome U74Av2 array (MG-U74Av2)
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2011-07-01 | E-GEOD-29848 | biostudies-arrayexpress
Project description:This analysis aims at finding the microRNAs present in three chick embryo undifferentiated tissues (pools of tissues): Undetermined Presomitic Mesoderm (PSM-U), Determined Presomitic Mesoderm (PSM-D) and Distal limb bud (Limb).
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