Cell Replating

Cell replating is a cell culture technique in which cells are detached from one culture surface and seeded onto a fresh substrate, allowing researchers to reset culture conditions and control cell density. The process typically involves enzymatic or mechanical dissociation, transfer into nutrient medium, and redistribution across a new surface to support renewed attachment and growth. In neuroscience, replating helps maintain neural cell cultures, generate experimental replicates, and examine how substrate, density, or treatment conditions influence cell survival, morphology, differentiation, and network formation. Consistent replating practices improve reproducibility in studies of neuronal development, cellular signaling, disease models, and neuropharmacology.

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JoVE EoE - Neuronal Culture Techniques

Replating Differentiated Neurons Derived from Human Pluripotent Stem Cells

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2025

The video showcases a method to detach and replant human pluripotent stem cell-derived neurons using a mild proteolytic enzyme. After filtration and centrifugation, the neurons are replated on a coated multi-well plate for incubation, promoting attachment and maturation.

Post-differentiation Replating of Human Pluripotent Stem Cell-derived Neurons for High-content Screening of Neuritogenesis and Synapse Maturation

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Cited by 9 •

2019

This protocol describes a detailed procedure for resuspending and culturing human stem cell derived neurons that were previously differentiated from neural progenitors in vitro for multiple weeks. The procedure facilitates imaging-based assays of neurites, synapses, and late-expressing neuronal markers in a format compatible with light microscopy and high-content screening.

Feeder-free Derivation of Neural Crest Progenitor Cells from Human Pluripotent Stem Cells

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Cited by 24 •

2014

Neural crest (NC) cells derived from human pluripotent stem cells (hPSC) have great potential for modeling human development and disease and for cell replacement therapies. Here, a feeder-free adaptation of the currently widely used in vitro differentiation protocol for the derivation of NC cells from hPSCs is presented.

Directed Dopaminergic Neuron Differentiation from Human Pluripotent Stem Cells

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Cited by 58 •

2014

We, based on knowledge from developmental biology and published research, developed an optimized protocol to efficiently generate A9 midbrain dopaminergic neurons from both human embryonic stem cells and human induced pluripotent stem cells, which would be useful for disease modeling and cell replacement therapy for Parkinson’s disease.

Feeder-free Derivation of Melanocytes from Human Pluripotent Stem Cells

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Cited by 12 •

2016

This work describes an in vitro differentiation protocol to produce pigmented, mature melanocytes from human pluripotent stem cells via a neural crest and melanoblast intermediate stage using a feeder-free, 25 day protocol.

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