title: Astroglial cells regulate the developmental timeline of human neurons differentiated from induced pluripotent stem cells.
abstract: Neurons derived from human induced-pluripotent stem cells (hiPSCs) have been used to model a variety of neurological disorders. Different protocols have been used to differentiate hiPSCs into neurons, but their functional maturation process has varied greatly among different studies. Here, we demonstrate that laminin, a commonly used substrate for iPSC cultures, was inefficient to promote fully functional maturation of hiPSC-derived neurons. In contrast, astroglial substrate greatly accelerated neurodevelopmental processes of hiPSC-derived neurons. We have monitored the neural differentiation and maturation process for up to two months after plating hiPSC-derived neuroprogenitor cells (hNPCs) on laminin or astrocytes. We found that one week after plating hNPCs, there were 21-fold more newly differentiated neurons on astrocytes than on laminin. Two weeks after plating hNPCs, there were 12-fold more dendritic branches in neurons cultured on astrocytes than on laminin. Six weeks after plating hNPCs, the Na(+) and K(+) currents, as well as glutamate and GABA receptor currents, were 3-fold larger in neurons cultured on astrocytes than on laminin. And two months after plating hNPCs, the spontaneous synaptic events were 8-fold more in neurons cultured on astrocytes than on laminin. These results highlight a critical role of astrocytes in promoting neural differentiation and functional maturation of human neurons derived from hiPSCs. Moreover, our data presents a thorough developmental timeline of hiPSC-derived neurons in culture, providing important benchmarks for future studies on disease modeling and drug screening.
authors.affiliation: Department of Biology, Huck Institutes of Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Tang
authors.forename: Li
authors.initials: L
authors.lastname: Zhou
authors.forename: Alecia M
authors.initials: AM
authors.lastname: Wagner
authors.forename: Maria C N
authors.initials: MC
authors.lastname: Marchetto
authors.forename: Alysson R
authors.initials: AR
authors.lastname: Muotri
authors.forename: Fred H
authors.initials: FH
authors.lastname: Gage
authors.forename: Gong
authors.initials: G
authors.lastname: Chen
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice; D009474:Neurons / Q000166:cytology*