Culture, differentiate, and mature neurons derived from human embryonic stem (ES) or induced pluripotent stem (iPS) cells by using a complete medium optimized to promote, rather than inhibit neuronal activity.
For your convenience, BrainPhys™ hPSC Neuron Kit includes serum-free BrainPhys™ Neuronal Medium (basal medium), supplements, and growth factors to enable you to generate and mature different neuronal subtypes from human ES/iPS cell-derived neural progenitor cells. Based on the formulation by Bardy and Gage (Bardy et al. PNAS, 2015), BrainPhys™ Neuronal Medium mimics the extracellular environment of the central nervous system (CNS) to yield a higher proportion of synaptically active neurons. Brewer’s B27-based (Brewer et al. J Neurosci Res., 1993) NeuroCult™ SM1 Neuronal Supplement ensures cell health and encourages neurite outgrowth and branching in short- and long-term serum-free cultures, and N2 Supplement-A supports the in vitro differentiation of ES/iPS-derived cells to neuronal subtypes. Included BDNF and GDNF growth factors support lineage-specific differentiation.
To avoid shocking your cells with media changes, you can also use BrainPhys™ medium when performing functional assays, such as microelectrode array-based recordings or live-fluorescent imaging.
View our additional resources to learn more about the BrainPhys™ system.
Subtype Basal Media, Specialized Media
Cell Type Neural Cells, PSC-Derived, Neurons, Pluripotent Stem Cells
Area of Interest Disease Modeling, Drug Discovery and Toxicity Testing, Neuroscience, Stem Cell Biology
Formulation Category Serum-Free
Data Figures
Table 1. Properties of Culture Media (C Bardy et al. Proc Natl Acad Sci USA, 2015)
Check-mark denotes physiological conditions and supported activities according to C Bardy et al. Proc Natl Acad Sci USA, 2015.
Figure 1. 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.
Figure 2. 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
Figure 3. 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.
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.
Modelling Lyssavirus Infections in Human Stem Cell-Derived Neural Cultures. V. Sundaramoorthy et al. Viruses 2020 mar
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.
Maturation of Human Pluripotent Stem Cell-Derived Cerebellar Neurons in the Absence of Co-culture. T. P. Silva et al. Frontiers in bioengineering and biotechnology 2020
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.
One-Stop Microfluidic Assembly of Human Brain Organoids To Model Prenatal Cannabis Exposure. Z. Ao et al. Analytical chemistry 2020
Abstract
Prenatal cannabis exposure (PCE) influences human brain development, but it is challenging to model PCE using animals and current cell culture techniques. Here, we developed a one-stop microfluidic platform to assemble and culture human cerebral organoids from human embryonic stem cells (hESC) to investigate the effect of PCE on early human brain development. By incorporating perfusable culture chambers, air-liquid interface, and one-stop protocol, this microfluidic platform can simplify the fabrication procedure and produce a large number of organoids (169 organoids per 3.5 cm × 3.5 cm device area) without fusion, as compared with conventional fabrication methods. These one-stop microfluidic assembled cerebral organoids not only recapitulate early human brain structure, biology, and electrophysiology but also have minimal size variation and hypoxia. Under on-chip exposure to the psychoactive cannabinoid, $\Delta$-9-tetrahydrocannabinol (THC), cerebral organoids exhibited reduced neuronal maturation, downregulation of cannabinoid receptor type 1 (CB1) receptors, and impaired neurite outgrowth. Moreover, transient on-chip THC treatment also decreased spontaneous firing in these organoids. This one-stop microfluidic technique enables a simple, scalable, and repeatable organoid culture method that can be used not only for human brain organoids but also for many other human organoids including liver, kidney, retina, and tumor organoids. This technology could be widely used in modeling brain and other organ development, developmental disorders, developmental pharmacology and toxicology, and drug screening.
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Legal Statement: BrainPhys is a registered trademark of the Salk Institute for Biological Studies, used under exclusive license. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
