title: Dynamic mass redistribution assay decodes differentiation of a neural progenitor stem cell.
abstract: Stem cells hold great potential in drug discovery and development. However, challenges remain to quantitatively measure the functions of stem cells and their differentiated products. Here, we applied fluorescent imaging, quantitative real-time PCR, and label-free dynamic mass redistribution (DMR) assays to characterize the differentiation process of the ReNcell VM human neural progenitor stem cell. Immunofluorescence imaging showed that after growth factor withdrawal, the neuroprogenitor stem cell was differentiated into dopaminergic neurons, astrocytes, and oligodendrocytes, thus creating a neuronal cell system. High-performance liquid chromatography analysis showed that the differentiated cell system released dopamine upon depolarization with KCl. In conjunction with quantitative real-time PCR, DMR assays using a G-protein-coupled receptor agonist library revealed that a subset of receptors, including dopamine D(1) and D(4) receptors, underwent marked alterations in both receptor expression and signaling pathway during the differentiation process. These findings suggest that DMR assays can decode the differentiation process of stem cells at the cell system level.
authors.affiliation: Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, USA.
authors.forename: Sadashiva
authors.initials: S
authors.lastname: Pai
authors.forename: Florence
authors.initials: F
authors.lastname: Verrier
authors.forename: Haiyan
authors.initials: H
authors.lastname: Sun
authors.forename: Haibei
authors.initials: H
authors.lastname: Hu
authors.forename: Ann M
authors.initials: AM
authors.lastname: Ferrie
authors.forename: Azita
authors.initials: A
authors.lastname: Eshraghi
authors.forename: Ye
authors.initials: Y
authors.lastname: Fang
mesh_terms: D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004298:Dopamine / Q000032:analysis; D059290:Dopaminergic Neurons / Q000166:cytology / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D057054:Molecular Imaging / Q000379:methods*; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D055495:Neurogenesis; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism; D011189:Potassium Chloride / Q000494:pharmacology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D017447:Receptors, Dopamine D1 / Q000378:metabolism; D050638:Receptors, Dopamine D4 / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D054852:Small Molecule Libraries / Q000494:pharmacology
chemical_list: C493599:DRD4 protein, human; D017447:Receptors, Dopamine D1; D043562:Receptors, G-Protein-Coupled; D054852:Small Molecule Libraries; D050638:Receptors, Dopamine D4; D011189:Potassium Chloride; D004298:Dopamine