title: Knocking down of heat-shock protein 27 directs differentiation of functional glutamatergic neurons from placenta-derived multipotent cells.
abstract: This study presents human placenta-derived multipotent cells (PDMCs) as a source from which functional glutamatergic neurons can be derived. We found that the small heat-shock protein 27 (HSP27) was downregulated during the neuronal differentiation process. The in vivo temporal and spatial profiles of HSP27 expression were determined and showed inverted distributions with neuronal proteins during mouse embryonic development. Overexpression of HSP27 in stem cells led to the arrest of neuronal differentiation; however, the knockdown of HSP27 yielded a substantially enhanced ability of PDMCs to differentiate into neurons. These neurons formed synaptic networks and showed positive staining for multiple neuronal markers. Additionally, cellular phenomena including the absence of apoptosis and rare proliferation in HSP27-silenced PDMCs, combined with molecular events such as cleaved caspase-3 and the loss of stemness with cleaved Nanog, indicated that HSP27 is located upstream of neuronal differentiation and constrains that process. Furthermore, the induced neurons showed increasing intracellular calcium concentrations upon glutamate treatment. These differentiated cells co-expressed the N-methyl-D-aspartate receptor, vesicular glutamate transporter, and synaptosomal-associated protein 25 but did not show expression of tyrosine hydroxylase, choline acetyltransferase or glutamate decarboxylase 67. Therefore, we concluded that HSP27-silenced PDMCs differentiated into neurons possessing the characteristics of functional glutamatergic neurons.
authors.affiliation: Department of Medical Research, Cathay General Hospital, Taipei, Taiwan.|Institute of Biomedical Engineering, National Central University, Jhongli, Taiwan.|Center for Biocellular Engineering, National Central University, Jhongli, Taiwan.|School of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan.
authors.forename: Yu-Che
authors.initials: YC
authors.lastname: Cheng
authors.affiliation: Department of Medical Research, Cathay General Hospital, Taipei, Taiwan.|Department of Biochemistry, National Defense Medical Center, Taipei, Taiwan.
authors.forename: Chi-Jung
authors.initials: CJ
authors.lastname: Huang
authors.affiliation: School of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan.
authors.forename: Yih-Jing
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: School of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan.
authors.forename: Lu-Tai
authors.initials: LT
authors.lastname: Tien
authors.affiliation: School of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan.
authors.forename: Wei-Chi
authors.initials: WC
authors.lastname: Ku
authors.affiliation: Department of Respiratory Therapy, Fu Jen Catholic University, New Taipei City, Taiwan.
authors.forename: Raymond
authors.initials: R
authors.lastname: Chien
authors.affiliation: Center for Biocellular Engineering, National Central University, Jhongli, Taiwan.|Department of Obstetrics and Gynecology, Cathay General Hospital, Taipei, Taiwan.
authors.forename: Fa-Kung
authors.initials: FK
authors.lastname: Lee
authors.affiliation: Center for Biocellular Engineering, National Central University, Jhongli, Taiwan.|Department of Anesthesiology, Cathay General Hospital, Taipei, Taiwan.
authors.forename: Chih-Cheng
authors.initials: CC
authors.lastname: Chien
mesh_terms: D002454:Cell Differentiation / Q000235:genetics*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005968:Glutamate Decarboxylase / Q000235:genetics; D005971:Glutamates / Q000235:genetics / Q000378:metabolism; D055551:HSP27 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D006801:Humans; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010920:Placenta / Q000166:cytology / Q000378:metabolism*; D011247:Pregnancy; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
chemical_list: D005971:Glutamates; D055551:HSP27 Heat-Shock Proteins; D014446:Tyrosine 3-Monooxygenase; D005968:Glutamate Decarboxylase