Hybrid Embryonic Stem Cell Corticogenesis

The hybrid male embryonic stem cell (ESC) lines were previously derived from blastocysts obtained from crosses between C57BL/J (B) and JF1 (J) mice³⁴ (link) and were maintained in gelatin-coated dishes with ESGRO complete plus medium (Millipore, SF001-500P) containing LIF (leukemia inhibitory factor), BMP4 (bone morphogenetic protein 4), and a GSK3-β (glycogen synthase kinase 3β) inhibitor. In vitro corticogenesis was performed as previously described,³⁵ (link) except that ESCs were plated on Matrigel-coated dishes (human ESC-qualified matrix, Corning), and that the defined default medium was supplemented with B27 (without vitamin A, Gibco) to improve cell survival and with 1 μM dorsomorphin homolog 1 (purified by C.C.H.) to promote neurogenesis.³⁶ (link) Using this protocol, neural precursor cells (NPCs) are the main cell population after 12 days (D12) of in vitro corticogenesis.³⁵ (link)

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Rengifo Rojas C., Cercy J., Perillous S., Gonthier-Guéret C., Montibus B., Maupetit-Méhouas S., Espinadel A., Dupré M., Hong C.C., Hata K., Nakabayashi K., Plagge A., Bouschet T., Arnaud P., Vaillant I, & Court F. (2024). Biallelic non-productive enhancer-promoter interactions precede imprinted expression of Kcnk9 during mouse neural commitment. Human Genetics and Genomics Advances, 5(2), 100271.

Publication 2024

SF001-500P B27 (without vitamin A,

Corresponding Organization : Université Clermont Auvergne

Other organizations : University of Maryland, Baltimore, National Center For Child Health and Development, Gunma University, University of Liverpool, Université de Montpellier, Institut de Génomique Fonctionnelle

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Variable analysis

independent variables

Crosses between C57BL/J (B) and JF1 (J) mice to derive hybrid male embryonic stem cell (ESC) lines
Plating ESCs on Matrigel-coated dishes (human ESC-qualified matrix, Corning)
Supplementing the defined default medium with B27 (without vitamin A, Gibco) to improve cell survival
Supplementing the defined default medium with 1 μM dorsomorphin homolog 1 (purified by C.C.H.) to promote neurogenesis

dependent variables

Neural precursor cells (NPCs) as the main cell population after 12 days (D12) of in vitro corticogenesis

control variables

Maintaining ESC lines in gelatin-coated dishes with ESGRO complete plus medium (Millipore, SF001-500P) containing LIF (leukemia inhibitory factor), BMP4 (bone morphogenetic protein 4), and a GSK3-β (glycogen synthase kinase 3β) inhibitor

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