title: Comparison of two types of non-adherent plate for neuronal differentiation of mouse embryonic stem cells.
abstract: In vitro differentiation systems of mouse embryonic stem cells (ESCs) are widely used as tools for studies of cell differentiation, organogenesis, and regenerative medicine. We have studied the regulation of neuron-specific imprinting genes, Ube3a and its antisense transcripts (Ube3a ATS), using in vitro neuronal differentiation of F1 hybrid ESCs. Each different non-adherent plate used for embryoid body (EB) formation during differentiation is associated with different costs; notably, plates coated with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer are more expensive than untreated polystyrene plates. Here, we assessed whether the polymer-coated plates gave better results than the untreated plates. The first stage of differentation was performed in the MPC polymer-coated or untreated plates. The formed EBs were then passaged onto laminin-coated plates for further differentiation into neurons. Neither the neuron-specific imprinting status of Ube3a nor the expression levels of the neuron-specific markers Ube3a ATS and Mtap2 differed between neurons prepared on untreated plates and those prepared on MPC polymer-coated plates. These results suggest that the two non-adherent plates displayed almost the same characteristics for inducing neuronal differentiation of mouse ESCs and EB formation. Our study proved that untreated polystyrene plates are a cost-effective choice for EB formation in in vitro differentiation systems of mouse ESCs.
authors.affiliation: Department of Environmental Medicine, Kochi Medical School, Kochi University, Oko-cho Kohasu, Nankoku, Kochi, 783-8505, Japan.
authors.forename: Masamitsu
authors.initials: M
authors.lastname: Eitoku
authors.affiliation: Department of Environmental Medicine, Kochi Medical School, Kochi University, Oko-cho Kohasu, Nankoku, Kochi, 783-8505, Japan.
authors.forename: Narufumi
authors.initials: N
authors.lastname: Suganuma
authors.affiliation: Department of Environmental Medicine, Kochi Medical School, Kochi University, Oko-cho Kohasu, Nankoku, Kochi, 783-8505, Japan. kiyosawa-fks@umin.org.
authors.forename: Hidenori
authors.initials: H
authors.lastname: Kiyosawa
keywords: Embryonic stem cell; Imprinting; M. musculus molossinus; Neuronal differentiation; Ube3a; Ube3a antisense