title: All-trans-retinoid acid induces the differentiation of encapsulated mouse embryonic stem cells into GABAergic neurons.
abstract: Embryonic stem (ES) cells are pluripotent cells that can differentiate into all three main germ layers: endoderm, mesoderm, and ectoderm. Although a number of methods have been developed to differentiate ES cells into neuronal phenotypes such as sensory and motor neurons, the efficient generation of GABAergic interneurons from ES cells still presents an ongoing challenge. Because the main output of inhibitory GABAergic interneurons is the gamma-aminobutyric-acid (GABA), a neurotransmitter whose controlled homeostasis is required for normal brain function, the efficient generation in culture of functional interneurons may have future implications on the treatment of neurological disorders such as epilepsy, autism, and schizophrenia. The goal of this work was to examine the generation of GABAergic neurons from mouse ES cells by comparing an embryoid body-based methodology versus a hydrogel-based encapsulation protocol that involves the use of all-trans-retinoid acid (RA). We observed that (1) there was a 2-fold increase in neuronal differentiation in encapsulated versus non-encapsulated cells and (2) there was an increase in the specificity for interneuronal differentiation in encapsulated cells, as assessed by mRNA expression and electrophysiology approaches. Furthermore, our results indicate that most of the neurons obtained from encapsulated mouse ES cells are GABA-positive (∼87%). Thus, these results suggest that combining encapsulation of ES cells and RA treatment provide a more efficient and scalable differentiation strategy for the generation in culture of functional GABAergic interneurons. This technology may have implications for future cell replacement therapies and the treatment of CNS disorders.
authors.affiliation: Department of Biological Sciences and Environmental Toxicology Program, Southern University and A&M College, Baton Rouge, LA 70813, USA.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Addae
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Yi
authors.forename: Ramkishore
authors.initials: R
authors.lastname: Gernapudi
authors.forename: Henrique
authors.initials: H
authors.lastname: Cheng
authors.forename: Alberto
authors.initials: A
authors.lastname: Musto
authors.forename: Eduardo
authors.initials: E
authors.lastname: Martinez-Ceballos
mesh_terms: D000818:Animals; D001921:Brain / Q000254:growth & development / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D064987:Cell- and Tissue-Based Therapy; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000637:transplantation; D059330:GABAergic Neurons / Q000166:cytology* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D053605:Leukemia Inhibitory Factor / Q000378:metabolism; D051379:Mice; D015221:Potassium Channels / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology*; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
chemical_list: D053605:Leukemia Inhibitory Factor; D015221:Potassium Channels; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D005680:gamma-Aminobutyric Acid; D014212:Tretinoin