Chat with an Expert

BrainPhys™ Neuronal Medium

Serum-free neurophysiological basal medium for improved neuronal function

BrainPhys™ Neuronal Medium

Serum-free neurophysiological basal medium for improved neuronal function

500 mL
Catalog #05790
87 USD

BrainPhys™ Neuronal Medium and SM1 Kit

Kit including BrainPhys™ Neuronal Medium and SM1 Neuronal Supplement for culture of primary and ES/iPS cell-derived neurons

500 mL Kit
Catalog #05792
169 USD

BrainPhys™ Neuronal Medium N2-A & SM1 Kit

Kit including BrainPhys™ Neuronal Medium, SM1 Neuronal Supplement, and N2 Supplement-A for culture of ES/iPS cell-derived neurons

500 mL Kit
Catalog #05793
251 USD

BrainPhys™ Primary Neuron Kit

Kit including BrainPhys™ Neuronal Medium, SM1 Neuronal Supplement and NeuroCult™ Neuronal Plating Medium for culture of primary-derived neurons

500 mL Kit
Catalog #05794
229 USD

BrainPhys™ hPSC Neuron Kit

Kit including BrainPhys™ Neuronal Medium, SM1 Neuronal Supplement, N2 Supplement-A, BDNF and GDNF for culture of ES/iPS cell-derived neurons

500 mL Kit
Catalog #05795
641 USD

Overview

BrainPhys™ Neuronal Medium is a serum-free neuronal basal medium. BrainPhys™ may be used to culture primary neurons or neurons derived from human pluripotent stem cells (hPSCs). Based on the formulation published by Cedric Bardy and Fred H. Gage (C Bardy et al. PNAS, 2015), BrainPhys™ is more representative of the central nervous system extracellular environment and increases the proportion of synaptically active neurons.

Applications of BrainPhys™ Neuronal Medium include culture of primary neurons, differentiation and maturation of hPSC-derived neurons, microelectrode array-based recording of neuronal activity, live fluorescent imaging (including calcium imaging and optogenetics) and transdifferentiation of somatic cells to neurons.

To ensure cell survival in long-term serum-free culture, BrainPhys™ must be combined with an appropriate supplement. For your convenience, various BrainPhys™ kits are available for primary or hPSC-derived neurons. BrainPhys™ Neuronal Medium and SM1 Kit (Catalog #05792) is recommended for culture of primary neurons. BrainPhys™ Primary Neuron Kit (Catalog #05794) is recommended for plating and culture of primary neurons. BrainPhys™ Neuronal Medium N2-A & SM1 Kit (Catalog #05793) and BrainPhys™ hPSC Neuron Kit (Catalog #05795) are recommended for the differentiation and maturation of hPSC-derived neurons, in combination with lineage-specific growth factors and/or small molecules (if necessary).
Advantages:
• More representative of the brain’s extracellular environment
• Improved neuronal function and a higher proportion of synaptically active neurons
• Perform functional assays without changing media and shocking cells
• Supports long-term culture of ES/iPS cell- and CNS-derived neurons
• Rigorous raw material screening and quality control ensure minimal lot-to-lot variability
Components:
  • BrainPhys™ Neuronal Medium and SM1 Kit (Catalog #05792)
    • BrainPhys™ Neuronal Medium, 500 mL (Catalog #05790)
    • NeuroCult™ SM1 Neuronal Supplement, 10 mL (Catalog #05711)
  • BrainPhys™ Neuronal Medium N2-A & SM1 Kit (Catalog #05793)
    • BrainPhys™ Neuronal Medium, 500 mL (Catalog #05790)
    • NeuroCult™ SM1 Neuronal Supplement, 10 mL (Catalog #05711)
    • N2 Supplement-A, 5 mL (Catalog #07152)
  • BrainPhys™ Primary Neuron Kit (Catalog #05794)
    • NeuroCult™ Neuronal Plating Medium, 100 mL (Catalog #05713)
    • NeuroCult™ SM1 Neuronal Supplement, 10 mL (Catalog #05711)
    • BrainPhys™ Neuronal Medium, 500 mL (Catalog #05790)
  • BrainPhys™ hPSC Neuron Kit (Catalog #05795)
    • BrainPhys™ Neuronal Medium, 500 mL (Catalog #05790)
    • NeuroCult™ SM1 Neuronal Supplement, 10 mL (Catalog #05711)
    • N2 Supplement-A, 5 mL (Catalog #07152)
    • Human Recombinant BDNF, 10 µg (Catalog #78005)
    • Human Recombinant GDNF, 10 µg (Catalog #78058)
Subtype:
Specialized Media; Basal Media
Cell Type:
Pluripotent Stem Cells; Neural Cells, PSC-Derived; Neurons
Species:
Human; Mouse; Rat
Application:
Cell Culture; Differentiation; Maintenance
Brand:
BrainPhys
Area of Interest:
Neuroscience; Stem Cell Biology; Disease Modeling; Drug Discovery and Toxicity Testing
Formulation:
Serum-Free; Defined

Scientific Resources

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.

Data and Publications

Data

Table 1. Properties of Culture Media (C Bardy et al. Proc Natl Acad Sci USA, 2015)

Check-mark denotes physiological conditions

Check-mark denotes physiological conditions and supported activities according to C Bardy et al. Proc Natl Acad Sci USA, 2015.

Rodent Neurons Matured in BrainPhys™ Neuronal Medium

Figure 1. Rodent Neurons Matured in BrainPhys™ Neuronal Medium are Healthy and Morphologically Mature

(A,C) Primary rat E18 cortical neurons were plated in NeuroCult™ Neuronal Basal Medium, supplemented with NeuroCult™ SM1 Neuronal Supplement. After 5 DIV, the cultures were transitioned to BrainPhys™ Neuronal Medium, supplemented with NeuroCult™ SM1, by performing half-medium changes every 3-4 days. Neurons were cultured for 14 (A) or 21 (C) DIV. (B,D) Primary rat E18 cortical neurons were plated and matured in a traditional neuronal medium (Neurobasal Medium), supplemented with NeuroCult™ SM1 Neuronal Supplement for 14 (B) or 21 (D) DIV. Neuronal morphology of BrainPhys™ Neuronal Medium-matured neurons is consistent with neurons plated and matured in a traditional neuronal medium.

Primary Neuronal Cultures Matured in BrainPhys™ Neuronal Medium Have Greater Numbers of Neurons

Figure 2. Primary Neuronal Cultures Matured in BrainPhys™ Neuronal Medium Have Greater Numbers of Neurons

Primary rat E18 cortical neurons were plated in either NeuroCult™ Neuronal Basal Medium (NCSM1) or Neurobasal Medium (NBSM1), supplemented with NeuroCult™ SM1. After 5 DIV, half of the cultures were transitioned to BrainPhys™ Neuronal Medium, supplemented with NeuroCult™ SM1, by performing half-medium changes every 3-4 days. The other half of the cultures were maintained in the same medium as used for plating. After 21 DIV, more neurons were evident in the cultures matured in BrainPhys™ Neuronal Medium, regardless of whether NeuroCult™ Neuronal Basal Medium or Neurobasal Medium was used as the plating medium. (n = 2, mean ± SEM [triplicate wells were set up for each experiment]).

Rodent Neuronal Cultures Matured in BrainPhys™ Neuronal Medium Show Improved Excitatory and Inhibitory Synaptic Activity

Figure 3. Rodent Neuronal Cultures Matured in BrainPhys™ Neuronal Medium Show Improved Excitatory and Inhibitory Synaptic Activity

(A,C) Primary rat E18 cortical neurons were plated in NeuroCult™ Neuronal Basal Medium, supplemented with NeuroCult™ SM1 Neuronal Supplement. After 5 DIV, the cultures were transitioned to BrainPhys™ Neuronal Medium, supplemented with NeuroCult™ SM1 Neuronal Supplement, by performing half-medium changes every 3 - 4 days. Neurons were cultured for 21 DIV. (B,D) Primary rat E18 cortical neurons were plated and matured in a traditional neuronal medium (Neurobasal Medium), supplemented with NeuroCult™ SM1 Neuronal Supplement for 21 DIV. (A,C) Neurons matured in BrainPhys™ Neuronal Medium showed spontaneous excitatory (AMPA-mediated; A) and inhibitory (GABA-mediated; C) synaptic events. The frequency and amplitude of spontaneous synaptic events is consistently greater in neuronal cultures matured in BrainPhys™ Neuronal Medium, compared to neurons plated and matured in a traditional neuronal medium (B,D). Traces are representative.

Expression of Pre-Synaptic Markers in Rodent Neurons Matured in BrainPhys™ Neuronal Medium

Figure 4. Expression of Pre-Synaptic Markers in Rodent Neurons Matured in BrainPhys™ Neuronal Medium

Primary rat E18 cortical neurons were plated in NeuroCult™ Neuronal Basal Medium, supplemented with NeuroCult™ SM1 Neuronal Supplement. After 5 DIV, the cultures were transitioned to BrainPhys™ Neuronal Medium, supplemented with NeuroCult™ SM1 Neuronal Supplement, by performing half-medium changes every 3 - 4 days. Neurons cultured for 21 DIV are phenotypically mature, as indicated by the presence of an extensive dendritic arbor. The pre-synaptic marker synapsin (A,B; green) is concentrated in discrete puncta distributed along the somata and dendritic processes, as defined by the dendritic marker MAP2 (A,C; red). Scale bar= 50 µm.

hPSC-Derived Neurons Generated in BrainPhys™ Neuronal Medium Express Markers of Neuronal Maturity After 14 and 44 Days of Differentiation

Figure 5. hPSC-Derived Neurons Generated in BrainPhys™ Neuronal Medium Express Markers of Neuronal Maturity After 14 and 44 Days of Differentiation

NPCs were generated from H9 cells using STEMdiff™ Neural Induction Medium in an embryoid body-based protocol. Next, NPCs were cultured in (A,C) BrainPhys™ Neuronal Medium, supplemented with 2% NeuroCult™ SM1 Supplement, 1% N2 Supplement-A, 20 ng/mL GDNF, 20 ng/mL BDNF, 1 mM db-cAMP and 200 nM ascorbic acid to initiate neuronal differentiation, or (B,D) DMEM/F12 under the same supplementation conditions. After 14 and 44 days of differentiation and maturation, neurons express the synaptic marker Synapsin 1 (green) and the mature neuronal marker MAP2 (red). In this example, neurons matured in BrainPhys™ Neuronal Medium show increased Synapsin 1 staining. Scale bar= 100 µm

hPSC-Derived Neurons Generated in BrainPhys™ Neuronal Medium and NeuroCult™ SM1 and N2 Supplements are Healthy and Morphologically Normal

Figure 6. hPSC-Derived Neurons Generated in BrainPhys™ Neuronal Medium and NeuroCult™ SM1 and N2 Supplements are Healthy and Morphologically Normal

NPCs were generated from H9 cells using STEMdiff™ Neural Induction Medium in an embryoid body-based protocol. Next, NPCs were cultured for 44 DIV in (A) BrainPhys™ Neuronal Medium, supplemented with 2% NeuroCult™ SM1 Supplement, 1% N2 Supplement-A, 20 ng/mL GDNF, 20 ng/mL BDNF, 1 mM db-cAMP and 200 nM ascorbic acid to initiate neuronal differentiation, or (B) DMEM/F12 under the same supplementation conditions. Neuronal cultures differentiated from NPCs in BrainPhys™ Neuronal Medium display extensive neurite outgrowth and reduced cellular debris compared to cultures differentiated in DMEM/F12. Scale bar= 100 µm.

hPSC-Derived Neurons Matured in BrainPhys™ Neuronal Medium Show Improved Excitatory and Inhibitory Synaptic Activity

Figure 7. hPSC-Derived Neurons Matured in BrainPhys™ Neuronal Medium Show Improved Excitatory and Inhibitory Synaptic Activity

NPCs were generated from H9 cells using STEMdiff™ Neural Induction Medium in an embryoid body-based protocol. Next, NPCs were cultured for 44 DIV in (A,C) BrainPhys™ Neuronal Medium, supplemented with 2% NeuroCult™ SM1 Supplement, 1% N2 Supplement-A, 20 ng/mL GDNF, 20 ng/mL BDNF, 1 mM db-cAMP and 200 nM ascorbic acid to initiate neuronal differentiation, or (B,D) in DMEM/F12 under the same supplementation conditions. (A,C) Neurons matured in BrainPhys™ Neuronal Medium showed spontaneous excitatory (AMPA-mediated; A) and inhibitory (GABA-mediated; C) synaptic events. The frequency and amplitude of spontaneous synaptic events is consistently greater in neuronal cultures matured in BrainPhys™ Neuronal Medium, compared to neurons plated and matured in DMEM/F12 (B,D). Traces are representative.

Publications

(28)
eLife 2018 MAY

A homozygous loss-of-function CAMK2A mutation causes growth delay, frequent seizures and severe intellectual disability.

P. H. Chia et al.

Abstract

Calcium/calmodulin-dependent protein kinase II (CAMK2) plays fundamental roles in synaptic plasticity that underlies learning and memory. Here, we describe a new recessive neurodevelopmental syndrome with global developmental delay, seizures and intellectual disability. Using linkage analysis and exome sequencing, we found that this disease maps to chromosome 5q31.1-q34 and is caused by a biallelic germline mutation in CAMK2A. The missense mutation, p.His477Tyr is located in the CAMK2A association domain that is critical for its function and localization. Biochemically, the p.His477Tyr mutant is defective in self-oligomerization and unable to assemble into the multimeric holoenzyme.In vivo, CAMK2AH477Y failed to rescue neuronal defects in C. elegans lacking unc-43, the ortholog of human CAMK2A. In vitro, neurons derived from patient iPSCs displayed profound synaptic defects. Together, our data demonstrate that a recessive germline mutation in CAMK2A leads to neurodevelopmental defects in humans and suggest that dysfunctional CAMK2 paralogs may contribute to other neurological disorders.
Scientific reports 2018 MAY

Acute Physiology and Neurologic Outcomes after Brain Injury in SCOP/PHLPP1 KO Mice.

T. C. Jackson et al.

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 MAY

Mitochondrial Aging Defects Emerge in Directly Reprogrammed Human Neurons due to Their Metabolic Profile.

Y. Kim et al.

Abstract

Mitochondria are a major target for aging and are instrumental in the age-dependent deterioration of the human brain, but studying mitochondria in aging human neurons has been challenging. Direct fibroblast-to-induced neuron (iN) conversion yields functional neurons that retain important signs of aging, in contrast to iPSC differentiation. Here, we analyzed mitochondrial features in iNs from individuals of different ages. iNs from old donors display decreased oxidative phosphorylation (OXPHOS)-related gene expression, impaired axonal mitochondrial morphologies, lower mitochondrial membrane potentials, reduced energy production, and increased oxidized proteins levels. In contrast, the fibroblasts from which iNs were generated show only mild age-dependent changes, consistent with a metabolic shift from glycolysis-dependent fibroblasts to OXPHOS-dependent iNs. Indeed, OXPHOS-induced old fibroblasts show increased mitochondrial aging features similar to iNs. Our data indicate that iNs are a valuable tool for studying mitochondrial aging and support a bioenergetic explanation for the high susceptibility of the brain to aging.
Cell reports 2018 JUN

CaMKII Metaplasticity Drives Abeta$ Oligomer-Mediated Synaptotoxicity.

P. Opazo et al.

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.
Current protocols in cell biology 2018 JUN

Transcription Factor-Mediated Differentiation of Human iPSCs into Neurons.

M. S. Fernandopulle et al.

Abstract

Accurate modeling of human neuronal cell biology has been a long-standing challenge. However, methods to differentiate human induced pluripotent stem cells (iPSCs) to neurons have recently provided experimentally tractable cell models. Numerous methods that use small molecules to direct iPSCs into neuronal lineages have arisen in recent years. Unfortunately, these methods entail numerous challenges, including poor efficiency, variable cell type heterogeneity, and lengthy, expensive differentiation procedures. We recently developed a new method to generate stable transgenic lines of human iPSCs with doxycycline-inducible transcription factors at safe-harbor loci. Using a simple two-step protocol, these lines can be inducibly differentiated into either cortical (i3 Neurons) or lower motor neurons (i3 LMN) in a rapid, efficient, and scalable manner (Wang et al., 2017). In this manuscript, we describe a set of protocols to assist investigators in the culture and genetic engineering of iPSC lines to enable transcription factor-mediated differentiation of iPSCs into i3 Neurons or i3 LMNs, and we present neuronal culture conditions for various experimental applications. {\textcopyright} 2018 by John Wiley & Sons, Inc.
STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.