title: Impedimetric real-time monitoring of neural pluripotent stem cell differentiation process on microelectrode arrays.
abstract: In today's neurodevelopment and -disease research, human neural stem/progenitor cell-derived networks represent the sole accessible in vitro model possessing a primary phenotype. However, cultivation and moreover, differentiation as well as maturation of human neural stem/progenitor cells are very complex and time-consuming processes. Therefore, techniques for the sensitive non-invasive, real-time monitoring of neuronal differentiation and maturation are highly demanded. Using impedance spectroscopy, the differentiation of several human neural stem/progenitor cell lines was analyzed in detail. After development of an optimum microelectrode array for reliable and sensitive long-term monitoring, distinct cell-dependent impedimetric parameters that could specifically be associated with the progress and quality of neuronal differentiation were identified. Cellular impedance changes correlated well with the temporal regulation of biomolecular progenitor versus mature neural marker expression as well as cellular structure changes accompanying neuronal differentiation. More strikingly, the capability of the impedimetric differentiation monitoring system for the use as a screening tool was demonstrated by applying compounds that are known to promote neuronal differentiation such as the γ-secretase inhibitor DAPT. The non-invasive impedance spectroscopy-based measurement system can be used for sensitive and quantitative monitoring of neuronal differentiation processes. Therefore, this technique could be a very useful tool for quality control of neuronal differentiation and moreover, for neurogenic compound identification and industrial high-content screening demands in the field of safety assessment as well as drug development.
authors.affiliation: Centre for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Seidel
authors.affiliation: Centre for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Obendorf
authors.affiliation: Centre for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Beate
authors.initials: B
authors.lastname: Englich
authors.affiliation: Centre for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Heinz-Georg
authors.initials: HG
authors.lastname: Jahnke
authors.affiliation: Institute of Reconstructive Neurobiology, University of Bonn and Hertie Foundation, Sigmund-Freud-Strasse 25, 53127 Bonn, Germany.
authors.forename: Vesselina
authors.initials: V
authors.lastname: Semkova
authors.affiliation: LIFE&BRAIN GmbH, Sigmund-Freud-Strasse 25, 53127 Bonn, Germany; Institute of Reconstructive Neurobiology, University of Bonn and Hertie Foundation, Sigmund-Freud-Strasse 25, 53127 Bonn, Germany.
authors.forename: Simone
authors.initials: S
authors.lastname: Haupt
authors.affiliation: CECS, I-STEM, AFM, Institute for Stem Cell Therapy and Exploration of Monogenic Diseases, Genopole Campus 1, 5 rue Henri Desbruères, 91030 Evry Cedex, France.
authors.forename: Mathilde
authors.initials: M
authors.lastname: Girard
authors.affiliation: INSERM U861, I-STEM, AFM, Institute for Stem Cell Therapy and Exploration of Monogenic Diseases, Genopole Campus 1, 5 rue Henri Desbruères, 91030 Evry Cedex, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Peschanski
authors.affiliation: LIFE&BRAIN GmbH, Sigmund-Freud-Strasse 25, 53127 Bonn, Germany; Institute of Reconstructive Neurobiology, University of Bonn and Hertie Foundation, Sigmund-Freud-Strasse 25, 53127 Bonn, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Brüstle
authors.affiliation: Centre for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany. Electronic address: andrea.robitzki@bbz.uni-leipzig.de.
authors.forename: Andrea A
authors.initials: AA
authors.lastname: Robitzki
mesh_terms: D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002478:Cells, Cultured; D058266:Dielectric Spectroscopy / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D008839:Microelectrodes; D058953:Neural Stem Cells / Q000166:cytology* / Q000502:physiology*; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D046888:Tissue Array Analysis / Q000295:instrumentation
keywords: Equivalent circuit modeling; Impedance spectroscopy; Label-free neuronal differentiation monitoring; Microelectrode array; Neuronal stem cells