Inducible Neurogenesis from Human iPSCs

The human iPSC cell line WTC11 carrying a doxycycline-inducible Ngn2 transgene (Miyaoka et al. 2014 (link); Wang et al. 2017 (link)) were cultivated as previously described (Bell et al. 2021 (link); Buchholz et al. 2022 ). Briefly, iPSCs were cultured on GelTrex™-coated plates (1X, ThermoFisher Scientific) in StemMACS™ iPS-Brew XF (Miltenyi). When reaching confluence, the cultures were passaged with Versene™ passaging solution (ThermoFisher Scientific) and seeded in thiazovivine-supplemented (Axon Medchem) iPS-Brew for 1 day. Cells were grown at 37 °C and 5% CO₂ in a humidified incubator.

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Bell-Simons M., Buchholz S., Klimek J, & Zempel H. (2023). Laser-Induced Axotomy of Human iPSC-Derived and Murine Primary Neurons Decreases Somatic Tau and AT8 Tau Phosphorylation: A Single-Cell Approach to Study Effects of Acute Axonal Damage. Cellular and Molecular Neurobiology, 43(7), 3497-3510.

Publication 2023

GelTrex™-coated plates StemMACS™ iPS-Brew XF

Axon Cell line Cells Doxycycline Human Ips 1 Ipscs Transgene Versene

Corresponding Organization :

Other organizations : University Hospital Cologne

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Variable analysis

independent variables

Doxycycline-inducible Ngn2 transgene

dependent variables

Not explicitly mentioned

control variables

Cultivation of human iPSC cell line WTC11 on GelTrex™-coated plates
Culture medium StemMACS™ iPS-Brew XF
Passaging with Versene™ solution
Thiazovivine-supplemented iPS-Brew for 1 day
Incubation at 37 °C and 5% CO2 in a humidified incubator

controls

Positive control: Not mentioned
Negative control: Not mentioned

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