NeuroCult™ SM1 Neuronal Supplement
Supplement for the serum-free culture of primary tissue-derived neurons and hPSC-derived neural progenitor cells
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NeuroCult™ SM1 Neuronal Supplement -
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Cell culture supplement
Overview
NeuroCult™ SM1 (STEMCELL Modified-1) Neuronal Supplement is a standardized serum-free supplement for the culture of primary tissue-derived neurons and human pluripotent stem cell (hPSC)-derived neural progenitor cells.
The NeuroCult™ SM1 formulation was developed based on the published formulation of Brewer’s B27 supplement (Brewer et al.). It has been optimized to reproducibly support increased survival and maturation of functional neurons in both short- and long-term cultures with minimal glial cell contamination (< 1% GFAP+). NeuroCult™ SM1 may be used in combination with BrainPhys™ Neuronal Medium (Catalog #05790) or as a serum-replacement supplement for various customizable applications.
NeuroCult™ SM1 is also a component of BrainPhys™ Neuronal Medium and SM1 Kit (Catalog #05792), BrainPhys™ Neuronal Medium N2-A & SM1 Kit (Catalog #05793), and BrainPhys™ Primary Neuron Kit (Catalog #05794). For further details and protocols, refer to the Product Information Sheet (PIS) for BrainPhys™ (Document #DX20519), available at www.stemcell.com or contact us to request a copy.
The NeuroCult™ SM1 formulation was developed based on the published formulation of Brewer’s B27 supplement (Brewer et al.). It has been optimized to reproducibly support increased survival and maturation of functional neurons in both short- and long-term cultures with minimal glial cell contamination (< 1% GFAP+). NeuroCult™ SM1 may be used in combination with BrainPhys™ Neuronal Medium (Catalog #05790) or as a serum-replacement supplement for various customizable applications.
NeuroCult™ SM1 is also a component of BrainPhys™ Neuronal Medium and SM1 Kit (Catalog #05792), BrainPhys™ Neuronal Medium N2-A & SM1 Kit (Catalog #05793), and BrainPhys™ Primary Neuron Kit (Catalog #05794). For further details and protocols, refer to the Product Information Sheet (PIS) for BrainPhys™ (Document #DX20519), available at www.stemcell.com or contact us to request a copy.
Advantages:
• Formulated to support improved cell survival in long-term primary neuronal culture
• Cultures feature increased neurite outgrowth and branching in short- and long-term cultures
• Product undergoes rigorous performance testing
• Cultures feature increased neurite outgrowth and branching in short- and long-term cultures
• Product undergoes rigorous performance testing
Contains:
• Antioxidants
• Vitamin A
• Insulin
• Other ingredients
• Vitamin A
• Insulin
• Other ingredients
Subtype:
Supplements
Cell Type:
Neural Cells, PSC-Derived; Neurons; Pluripotent Stem Cells
Species:
Human; Mouse; Rat
Application:
Cell Culture; Differentiation; Maintenance
Brand:
NeuroCult
Area of Interest:
Neuroscience; Stem Cell Biology
Formulation:
Serum-Free
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Scientific Resources
Product Documentation
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Educational Materials
(12)
Product Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
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Data and Publications
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Figure 1. Protocol for Plating and Culturing Primary Neurons with the SM1 Culture System
Primary rodent tissue dissociated in papain was plated in NeuroCult™ Neuronal Plating Medium, supplemented with NeuroCult™ SM1 Neuronal Supplement, L-Glutamine, and L-Glutamic Acid. On day 5, primary neurons were transitioned to BrainPhys™ Neuronal Medium, supplemented with NeuroCult™ SM1 Neuronal Supplement, by performing half-medium changes every 3 - 4 days.
Figure 2. The SM1 Culture System Supports Long-Term Culture of Rodent Neurons
Primary E18 rat cortical neurons were cultured in the SM1 Culture System. A large number of viable neurons are visible after (A) 21 and (B) 35 days, as demonstrated by their bright neuronal cell bodies, and extensive neurite outgrowth and branching. Neurons are evenly distributed over the culture surface with minimal cell clumping.
Figure 3. Pre- and Post-Synaptic Markers are Expressed in Rodent Neurons Cultured in the SM1 Culture System
Primary E18 rat cortical neurons were cultured in the SM1 Culture System. At 21 DIV, neurons are phenotypically mature, as indicated by the presence of an extensive dendritic arbor, and appropriate expression and localization of pre-synaptic synapsin (A,C; green) and post-synaptic PSD-95 (A,B; red) markers. Synapsin is concentrated in discrete puncta distributed along the somata and dendritic processes, as defined by the dendritic marker MAP2 (A,D; blue).
Figure 4. The SM1 Culture System Supports Increased Cell Survival
(A) Primary E18 rat cortical neurons were cultured in the SM1 Culture System or a Competitor Culture System for 21 days. Neurons cultured in the SM1 Culture System have a significantly higher number of viable cells compared to the competitor culture system (n = 4; mean ± 95% CI; *p < 0.05). (B) Primary E18 rat cortical neurons were cultured in Neurobasal® supplemented with NeuroCult™ SM1 Neuronal Supplement (SM1) or competitor B27-like supplements (Competitor 1,2,3) for 21 days. Cultures supplemented with NeuroCult™ SM1 Neuronal Supplement have an equal number of neurons compared to competitor-supplemented cultures. Bars represent standard error of mean.
Publications
(50)
Viruses 2020 mar
Modelling Lyssavirus Infections in Human Stem Cell-Derived Neural Cultures.
Abstract
Abstract
Rabies is a zoonotic neurological infection caused by lyssavirus that continues to result in devastating loss of human life. Many aspects of rabies pathogenesis in human neurons are not well understood. Lack of appropriate ex-vivo models for studying rabies infection in human neurons has contributed to this knowledge gap. In this study, we utilize advances in stem cell technology to characterize rabies infection in human stem cell-derived neurons. We show key cellular features of rabies infection in our human neural cultures, including upregulation of inflammatory chemokines, lack of neuronal apoptosis, and axonal transmission of viruses in neuronal networks. In addition, we highlight specific differences in cellular pathogenesis between laboratory-adapted and field strain lyssavirus. This study therefore defines the first stem cell-derived ex-vivo model system to study rabies pathogenesis in human neurons. This new model system demonstrates the potential for enabling an increased understanding of molecular mechanisms in human rabies, which could lead to improved control methods.
Frontiers in bioengineering and biotechnology 2020
Maturation of Human Pluripotent Stem Cell-Derived Cerebellar Neurons in the Absence of Co-culture.
Abstract
Abstract
The cerebellum plays a critical role in all vertebrates, and many neurological disorders are associated with cerebellum dysfunction. A major limitation in cerebellar research has been the lack of adequate disease models. As an alternative to animal models, cerebellar neurons differentiated from pluripotent stem cells have been used. However, previous studies only produced limited amounts of Purkinje cells. Moreover, in vitro generation of Purkinje cells required co-culture systems, which may introduce unknown components to the system. Here we describe a novel differentiation strategy that uses defined medium to generate Purkinje cells, granule cells, interneurons, and deep cerebellar nuclei projection neurons, that self-formed and differentiated into electrically active cells. Using a defined basal medium optimized for neuronal cell culture, we successfully promoted the differentiation of cerebellar precursors without the need for co-culturing. We anticipate that our findings may help developing better models for the study of cerebellar dysfunctions, while providing an advance toward the development of autologous replacement strategies for treating cerebellar degenerative diseases.
American journal of human genetics 2019
Mutations in ACTL6B Cause Neurodevelopmental Deficits and Epilepsy and Lead to Loss of Dendrites in Human Neurons.
Abstract
Abstract
We identified individuals with variations in ACTL6B, a component of the chromatin remodeling machinery including the BAF complex. Ten individuals harbored bi-allelic mutations and presented with global developmental delay, epileptic encephalopathy, and spasticity, and ten individuals with de novo heterozygous mutations displayed intellectual disability, ambulation deficits, severe language impairment, hypotonia, Rett-like stereotypies, and minor facial dysmorphisms (wide mouth, diastema, bulbous nose). Nine of these ten unrelated individuals had the identical de novo c.1027G{\textgreater}A (p.Gly343Arg) mutation. Human-derived neurons were generated that recaptured ACTL6B expression patterns in development from progenitor cell to post-mitotic neuron, validating the use of this model. Engineered knock-out of ACTL6B in wild-type human neurons resulted in profound deficits in dendrite development, a result recapitulated in two individuals with different bi-allelic mutations, and reversed on clonal genetic repair or exogenous expression of ACTL6B. Whole-transcriptome analyses and whole-genomic profiling of the BAF complex in wild-type and bi-allelic mutant ACTL6B neural progenitor cells and neurons revealed increased genomic binding of the BAF complex in ACTL6B mutants, with corresponding transcriptional changes in several genes including TPPP and FSCN1, suggesting that altered regulation of some cytoskeletal genes contribute to altered dendrite development. Assessment of bi-alleic and heterozygous ACTL6B mutations on an ACTL6B knock-out human background demonstrated that bi-allelic mutations mimic engineered deletion deficits while heterozygous mutations do not, suggesting that the former are loss of function and the latter are gain of function. These results reveal a role for ACTL6B in neurodevelopment and implicate another component of chromatin remodeling machinery in brain disease.
Scientific reports 2018 MAY
Acute Physiology and Neurologic Outcomes after Brain Injury in SCOP/PHLPP1 KO Mice.
Abstract
Abstract
Suprachiasmatic nucleus circadian oscillatory protein (SCOP) (a.k.a. PHLPP1) regulates long-term memory consolidation in the brain. Using a mouse model of controlled cortical impact (CCI) we tested if (1) brain tissue levels of SCOP/PHLPP1 increase after a traumatic brain injury (TBI), and (2) if SCOP/PHLPP1 gene knockout (KO) mice have improved (or worse) neurologic outcomes. Blood chemistry (pH, pCO2, pO2, pSO2, base excess, sodium bicarbonate, and osmolarity) and arterial pressure (MAP) differed in isoflurane anesthetized WT vs. KOs at baseline and up to 1 h post-injury. CCI injury increased cortical/hippocampal SCOP/PHLPP1 levels in WTs 7d and 14d post-injury. Injured KOs had higher brain tissue levels of phosphorylated AKT (pAKT) in cortex (14d post-injury), and higher levels of phosphorylated MEK (pMEK) in hippocampus (7d and 14d post-injury) and in cortex (7d post-injury). Consistent with an important role of SCOP/PHLPP1 on memory function, injured-KOs had near normal performance on the probe trial of the Morris water maze, whereas injured-WTs were impaired. CA1/CA3 hippocampal survival was lower in KOs vs. WTs 24 h post-injury but equivalent by 7d. No difference in 21d cortical lesion volume was detected. SCOP/PHLPP1 overexpression in cultured rat cortical neurons had no effect on 24 h cell death after a mechanical stretch-injury.
Cell reports 2018 JUN
CaMKII Metaplasticity Drives Abeta$ Oligomer-Mediated Synaptotoxicity.
Abstract
Abstract
Alzheimer's disease (AD) is emerging as a synaptopathology driven by metaplasticity. Indeed, reminiscent of metaplasticity, oligomeric forms of the amyloid-beta$ peptide (oAbeta$) prevent induction of long-term potentiation (LTP) via the prior activation of GluN2B-containing NMDA receptors (NMDARs). However, the downstream Ca2+-dependent signaling molecules that mediate aberrant metaplasticity are unknown. In this study, we show that oAbeta$ promotes the activation of Ca2+/calmodulin-dependent kinase II (CaMKII) via GluN2B-containing NMDARs. Importantly, we find that CaMKII inhibition rescues both the LTP impairment and the dendritic spine loss mediated by oAbeta$. Mechanistically resembling metaplasticity, oAbeta$ prevents subsequent rounds of plasticity from inducing CaMKII T286 autophosphorylation, as well as the associated anchoring and accumulation of synaptic AMPA receptors (AMPARs). Finally, prolonged oAbeta$ treatment-induced CaMKII misactivation leads to dendritic spine loss via the destabilization of surface AMPARs. Thus, our study demonstrates that oAbeta$ engages synaptic metaplasticity via aberrant CaMKII activation.
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