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Deep sequencing identify small molecule modulated microRNAs in the J1 mESCs

ABSTRACT: Many small molecular compounds reported were involved in improving reprogramming efficiency during inducing pluripotent stem cells (iPSC) or maintain a blastocyst-like state in embryonic stem cells (ESC), including BIO, CHIR99021 (CHIR), and vitamin C et al. However, the knowledge about these small molecules regulating miRNAs in ESC was limited. To understand the role of miRNAs during small molecules induced ESC maintenance and gain an insight how these small molecules regulates miRNAs expression; we performed small RNA sequencing using Illumina HiSeq 2000 under the compounds treatment.The data show the miRNAs regulated by SB431542

ORGANISM(S): Mus musculus

PROVIDER: GSE56385 | GEO | 2015/06/01

SECONDARY ACCESSION(S): PRJNA243083

REPOSITORIES: GEO

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Deep sequencing identifies small molecule modulated microRNAs in the J1 mESCs

Project description:Many small molecular compounds reported were involved in improving reprogramming efficiency during inducing pluripotent stem cells (iPSC) or maintain a blastocyst-like state in embryonic stem cells (ESC), including BIO, CHIR99021 (CHIR), and vitamin C et al. However, the knowledge about these small molecules regulating miRNAs in ESC was limited. To understand the role of miRNAs during small molecules induced ESC maintenance and gain an insight how these small molecules regulates miRNAs expression; we performed small RNA sequencing using Illumina HiSeq 2000 under the compounds treatment. Retinoic acid (RA), a stimulus molecule for cellular differentiation, is set as negative control. The data show the miRNAs regulated by BIO, CHIR, vitamin C and RA.

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Project description:Here, we report synergistic inhibition of glycogen synthase kinase 3 (GSK3), transforming growth factor β (TGF β), and Notch signaling pathways by small molecules can efficiently convert monolayer cultured hESCs into homogenous primitive neuroepithelium within one week under chemically defined condition. These primitive neuroepithelia can stably self-renew in the presence of leukemia inhibitory factor, GSK3 inhibitor (CHIR99021) and TGF β receptor inhibitor (SB431542); retain high neurogenic potential and responsiveness to instructive neural patterning cues toward midbrain and hindbrain neuronal subtypes; and exhibit in vivo integration. hESCs at about 20% confluence were treated with 3 μM CHIR99021, 2 μM SB431542, 0.1 μM Compound E (γ-Secretase Inhibitor XXI) in neural induction media containing Advanced DMEM/F12:Neurobasal (1:1), 1xN2, 1xB27, 1% Glutmax, 5 μg/ml BSA and 10 ng/ml hLIF, for 7 days. The culture was then split 1:3 for the next six passages using Accutase and cultured in neural induction media supplemented with 3 μM CHIR99021 and 2 μM SB431542 on X-ray inactivated MEF feeders or Matrigel-coated plates. After six passages, the cells were split 1:10 regularly. Global gene expression analysis of primitive neural stem cells

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