BrainPhys™ hPSC Neuron Kit
Kit for serum-free culture and differentiation of ES/iPS cell-derived neurons in BrainPhys™ Neuronal Medium
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What Our Scientist Says
I want to help neuroscientists like you create more physiological culture conditions, for more active and healthy neuronal cultures.
Overview
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 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.
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.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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.
Resources and Publications
Educational Materials (23)
Publications (51)
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Legal Statement:
BrainPhys is a registered trademark of the Salk Institute for Biological Studies, used under exclusive license.
Quality Statement:
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.
