ArticleStem cell research & therapy2015
A puromycin selectable cell line for the enrichment of mouse embryonic stem cell-derived V3 interneurons.
Article in Stem cell research & therapy, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Roles of spinal V3 interneurons: Roles in controlling movement in healthy and injured conditions.Neural regeneration research · 2026Article
- A transgenic mouse embryonic stem cell line for puromycin selection of V0Stem cell research & therapy · 2022Article
- Induction of Ventral Spinal V0 Interneurons from Mouse Embryonic Stem Cells.Stem cells and development · 2021Article
- Preparation of Neural Stem Cells and Progenitors: Neuronal Production and Grafting Applications.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Article
- Transplanting Cells for Spinal Cord Repair: Who, What, When, Where and Why?Cell transplantation · 2019Article
- Role of Propriospinal Neurons in Control of Respiratory Muscles and Recovery of Breathing Following Injury.Frontiers in systems neuroscience · 2019Review
- Derivation of Specific Neural Populations From Pluripotent Cells for Understanding and Treatment of Spinal Cord Injury.Developmental dynamics : an official publication of the American Association of Anatomists · 2019Review
- The Neuroplastic and Therapeutic Potential of Spinal Interneurons in the Injured Spinal Cord.Trends in neurosciences · 2018Review
- Genome engineering for CNS injury and disease.Current opinion in biotechnology · 2018Review
- Stem cells for spinal cord injury: Strategies to inform differentiation and transplantation.Biotechnology and bioengineering · 2017Review
- MicroRNA-363 and GATA-1 are regulated by HIF-1α in K562 cells under hypoxia.Molecular medicine reports · 2016Article
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4 authors.
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Abstract
introductionSpinal V3 interneurons (INs) are a commissural, glutamatergic, propriospinal neuron population that holds great potential for understanding locomotion circuitry and local rewiring after spinal cord injury. Embryonic stem cells hold promise as a cell source. However, the inevitable heterogeneity resulting from differentiation protocols makes studying post-mitotic stem cell-derived neuron populations difficult because proliferative glia quickly overtake a culture. Previously, an induction protocol for V3 INs was established. However, because of the heterogeneous population resulting from the induction protocol, functional characterization of the induced cells was not possible.
methodsA selectable murine transgenic embryonic stem cell (ESC) line (Sim1-Puro) was generated by recombineering. The expression of the puromycin resistance enzyme, puromycin N-acetyl-transferase (PAC), was knocked into the locus of a post-mitotic V3 IN marker (Sim1), allowing Sim1 gene regulatory elements to control PAC expression. The resulting cell line was characterized for Sim1 expression by in situ hybridization, for glutamatergic marker expression by immunocytochemistry and quantitative real time polymerase chain reaction (qRT-PCR), and for functional maturation by electrophysiology.
resultsPuromycin selection significantly enriched the population for V3 INs, allowing long-term characterization. The selected population expressed the neuronal marker β-III tubulin and the glutamatergic neuron marker VGluT2. The selected V3 INs also exhibited appropriate functional maturation, as assessed by electrophysiology, and remained glutamatergic for 2 weeks.
conclusionThe Sim1-Puro cell line provides a simple, high throughput method for generating large numbers of V3 INs from mouse ESCs for future in vitro and cell transplantation studies.
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