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
94 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
181 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
269 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
238 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
667 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:
Basal Media; Specialized Media
Cell Type:
Neural Cells, PSC-Derived; Neurons; Pluripotent Stem Cells
Species:
Human; Mouse; Rat
Application:
Cell Culture; Differentiation; Maintenance
Brand:
BrainPhys
Area of Interest:
Disease Modeling; Drug Discovery and Toxicity Testing; Neuroscience; Stem Cell Biology
Formulation:
Serum-Free

Scientific Resources

Educational Materials

(13)
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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. 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.

Rodent Neurons Matured in BrainPhys™ Neuronal Medium

Figure 2. Protocol for Culturing hPSCs with the SM1 Culture System

hPSCs were maintained in mTeSR™1 medium and then differentiated using the STEMdiff™ SMADi Neural Induction Kit. Following plating on PLO/laminin, half-medium changes were performed to transition to BrainPhys™ Neuronal Medium for maturation and long-term culture.

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

Figure 3. 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.

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

Figure 4. 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).

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

Figure 5. 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.

Rodent Neurons Matured in BrainPhys™ Neuronal Medium

Figure 6. 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 (product superseded with NeuroCult™ Neuronal Plating Medium which is a part of the BrainPhys™ Primary Neuron Kit), 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 competitor neuronal medium (Neurobasal®), 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 competitor neuronal medium (B,D). Traces are representative.

Rodent Neurons Matured in BrainPhys™ Neuronal Medium

Figure 7. Primary Neuronal Cultures Matured in BrainPhys™ Neuronal Medium Show Improved Electrical Activity in Microelectrode-Array Systems

Primary rat E18 cortical neurons were plated in a competitor neuronal medium (Neurobasal®) supplemented with NeuroCult™ SM1 Neuronal Supplement. After 5 DIV, half of the cultures were transitioned to BrainPhys™ Neuronal Medium, supplemented with NeuroCult™ SM1 Neuronal Supplement, by performing half-medium changes every 3 - 4 days. The other half of the cultures were maintained in the competitor neuronal medium throughout. The electrical activities of the neuronal cultures were measured twice a week using a microelectrode array (MEA) system (Axion Biosystems). (A) The mean firing rate of neurons cultured in BrainPhys™ Neuronal Medium increases over time, whereas the mean firing rate of neurons in the competitor neuronal medium condition remains low (n = 1; mean ± SEM, 128 electrodes). (B) The percentage of active electrodes (>0.005 Hz) of neurons matured in BrainPhys™ Neuronal Medium increases from 24% on day 14 to 69% on day 21, and then remains stable at 60 – 70% from days 21 – 44. In contrast, < 5% of electrodes was active in the competitor neuronal medium condition over the same 6-week period.

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

Figure 8. 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 9. 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 10. 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

(38)
Advanced healthcare materials 2019 oct

Soluble Signals and Remodeling in a Synthetic Gelatin-Based Hematopoietic Stem Cell Niche.

A. E. Gilchrist et al.

Abstract

Hematopoietic stem cells (HSCs) reside in the bone marrow within niches that provide microenvironmental signals in the form of biophysical cues, bound and diffusible biomolecules, and heterotypic cell-cell interactions that influence HSC fate decisions. This study seeks to inform the development of a synthetic culture platform that promotes ex vivo HSC expansion without exhaustion. A library of methacrylamide-functionalized gelatin (GelMA) hydrogels is used to explore remodeling and crosstalk from mesenchymal stromal cells (MSCs) on the expansion and quiescence of murine HSCs. The use of a degradable GelMA hydrogel enables MSC-mediated remodeling, yielding dynamic shifts in the matrix environment over time. An initially low-diffusivity hydrogel for co-culture of hematopoietic stem and progenitor cells to MSCs facilitates maintenance of an early progenitor cell population over 7 days. Excitingly, this platform promotes retention of a quiescent HSC population compared to HSC monocultures. These studies reveal MSC-density-dependent upregulation of MMP-9 and changes in hydrogel mechanical properties ($\Delta$E = 2.61 ± 0.72) suggesting MSC-mediated matrix remodeling may contribute to a dynamic culture environment. Herein, a 3D hydrogel is reported for ex vivo HSC culture, in which HSC expansion and quiescence is sensitive to hydrogel properties, MSC co-culture, and MSC-mediated hydrogel remodeling.
Scientific reports 2019 oct

Small extracellular vesicles convey the stress-induced adaptive responses of melanoma cells.

M. Harmati et al.

Abstract

Exosomes are small extracellular vesicles (sEVs), playing a crucial role in the intercellular communication in physiological as well as pathological processes. Here, we aimed to study whether the melanoma-derived sEV-mediated communication could adapt to microenvironmental stresses. We compared B16F1 cell-derived sEVs released under normal and stress conditions, including cytostatic, heat and oxidative stress. The miRNome and proteome showed substantial differences across the sEV groups and bioinformatics analysis of the obtained data by the Ingenuity Pathway Analysis also revealed significant functional differences. The in silico predicted functional alterations of sEVs were validated by in vitro assays. For instance, melanoma-derived sEVs elicited by oxidative stress increased Ki-67 expression of mesenchymal stem cells (MSCs); cytostatic stress-resulted sEVs facilitated melanoma cell migration; all sEV groups supported microtissue generation of MSC-B16F1 co-cultures in a 3D tumour matrix model. Based on this study, we concluded that (i) molecular patterns of tumour-derived sEVs, dictated by the microenvironmental conditions, resulted in specific response patterns in the recipient cells; (ii) in silico analyses could be useful tools to predict different stress responses; (iii) alteration of the sEV-mediated communication of tumour cells might be a therapy-induced host response, with a potential influence on treatment efficacy.
Mucosal immunology 2019 nov

NK cell recruitment limits tissue damage during an enteric helminth infection.

M. E. Gentile et al.

Abstract

Parasitic helminths cause significant damage as they migrate through host tissues to complete their life cycle. While chronic helminth infections are characterized by a well-described Type 2 immune response, the early, tissue-invasive stages are not well understood. Here we investigate the immune pathways activated during the early stages of Heligmosomoides polygyrus bakeri (Hpb), a natural parasitic roundworm of mice. In contrast to the Type 2 immune response present at later stages of infection, a robust Type 1 immune signature including IFNg production was dominant at the time of parasite invasion and granuloma formation. This early response was associated with an accumulation of activated Natural Killer (NK) cells, with no increase of other innate lymphoid cell populations. Parabiosis and confocal microscopy studies indicated that NK cells were recruited from circulation to the small intestine, where they surrounded parasitic larvae. NK cell recruitment required IFN$\gamma$ receptor signaling, but was independent of CXCR3 expression. The depletion of tissue-infiltrating NK cells altered neither worm burden nor parasite fitness, but increased vascular injury, suggesting a role for NK cells in mediating tissue protection. Together, these data identify an unexpected role for NK cells in promoting disease tolerance during the invasive stage of an enteric helminth infection.
Nature Neuroscience 2019 apr

Cerebral organoids at the air–liquid interface generate diverse nerve tracts with functional output

S. L. Giandomenico et al.

Abstract

Neural organoids have the potential to improve our understanding of human brain development and neurological disorders. However, it remains to be seen whether these tissues can model circuit formation with functional neuronal output. Here we have adapted air–liquid interface culture to cerebral organoids, leading to improved neuronal survival and axon outgrowth. The resulting thick axon tracts display various morphologies, including long-range projection within and away from the organoid, growth-cone turning, and decussation. Single-cell RNA sequencing reveals various cortical neuronal identities, and retrograde tracing demonstrates tract morphologies that match proper molecular identities. These cultures exhibit active neuronal networks, and subcortical projecting tracts can innervate mouse spinal cord explants and evoke contractions of adjacent muscle in a manner dependent on intact organoid-derived innervating tracts. Overall, these results reveal a remarkable self-organization of corticofugal and callosal tracts with a functional output, providing new opportunities to examine relevant aspects of human CNS development and disease.
Nature communications 2019

Human genome-edited hematopoietic stem cells phenotypically correct Mucopolysaccharidosis type I.

N. Gomez-Ospina et al.

Abstract

Lysosomal enzyme deficiencies comprise a large group of genetic disorders that generally lack effective treatments. A potential treatment approach is to engineer the patient's own hematopoietic system to express high levels of the deficient enzyme, thereby correcting the biochemical defect and halting disease progression. Here, we present an efficient ex vivo genome editing approach using CRISPR-Cas9 that targets the lysosomal enzyme iduronidase to the CCR5 safe harbor locus in human CD34+ hematopoietic stem and progenitor cells. The modified cells secrete supra-endogenous enzyme levels, maintain long-term repopulation and multi-lineage differentiation potential, and can improve biochemical and phenotypic abnormalities in an immunocompromised mouse model of Mucopolysaccharidosis type I. These studies provide support for the development of genome-edited CD34+ hematopoietic stem and progenitor cells as a potential treatment for Mucopolysaccharidosis type I. The safe harbor approach constitutes a flexible platform for the expression of lysosomal enzymes making it applicable to other lysosomal storage disorders.
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