Generation of Schwann Cells

Overview Materials and Reagents Steps Quality Control Troubleshooting Related Services

Schwann cells are the principal glial cells of the peripheral nervous system (PNS), responsible for axonal myelination and trophic support. As a critical component of nerve regeneration studies, disease modeling, and cell therapy, Schwann cells derived from induced pluripotent stem cells (iPSCs) provide an unlimited, patient-specific, and ethically acceptable cell source. Creative Biolabs offers state-of-the-art custom iPSC differentiation services, enabling researchers to obtain functionally mature Schwann cells with high purity and consistency.

This protocol outlines a step-by-step strategy to generate Schwann cells from iPSCs using a staged differentiation approach that mimics embryonic development, transitioning through neural crest intermediates. The methodology is optimized for reproducibility, scalability, and downstream applications in drug screening, disease modeling, and regenerative medicine.

Overview of the Generation of Schwann Cells

The generation of Schwann cells from human iPSCs is a multi-stage, lineage-guided differentiation process that closely recapitulates peripheral nervous system development in vivo. Schwann cells originate from neural crest cells (NCCs), a transient multipotent cell population arising during embryogenesis. Thus, to faithfully mimic this developmental trajectory, iPSCs must be induced through a neural lineage, transitioned into NCCs, and finally matured into Schwann cells using specific biochemical cues.

Fig. 1 Protocol for Schwann cell differentiation from iPSCs.^1,2

This process yields functionally relevant Schwann cells expressing lineage-specific markers (S100β, MBP, GFAP) and displaying key physiological behaviors such as myelination and neurotrophic support.

Importantly, iPSC-derived Schwann cells offer several advantages over primary cells.

Scalability: Unlimited expansion potential from renewable iPSC sources
Consistency: Controlled differentiation under defined conditions
Disease modeling: Access to patient-specific or gene-edited iPSC lines
Translational potential: Compatibility with drug screening and regenerative medicine

By leveraging Creative Biolabs' deep expertise in stem cell biology and neural differentiation, researchers gain access to a powerful platform for modeling peripheral neuropathies, screening neuroprotective compounds, and advancing cell-based therapies.

Materials and Reagents

Category	Item Description
iPSC culture	Matrigel, mTeSR1 medium, Accutase
Induction medium	Neurobasal medium, DMEM/F12, N2 supplement, B27 supplement
Signaling modulators	CHIR99021 (Wnt activator), SB431542 (TGF-β inhibitor), RA
Schwann differentiation	Heregulin-1β (NRG1), forskolin, PDGF-BB, IGF-1, BDNF
Coating materials	Poly-L-ornithine (PLO), laminin
Antibodies (validation)	S100β, p75NTR, SOX10, GFAP, MBP
Others	Penicillin/Streptomycin, PBS, Trypan blue, FBS

Protocol Steps

Maintenance of iPSCs

Culture iPSCs on Matrigel-coated plates using mTeSR1. Passage every 5–6 days using Accutase. Ensure 70–80% confluency and healthy colonies before initiating differentiation.

Neural Induction

Replace mTeSR1 with neural induction medium: DMEM/F12 + N2 + B27 + CHIR99021 + SB431542. Feed cells daily and monitor morphology—neuroepithelial rosettes should appear around Day 5. Optional: Apply dual-SMAD inhibition (LDN193189 may be added) for enhanced neural lineage commitment.

Neural Crest Cell Differentiation

Transition to NCC induction medium: Neurobasal + N2 + B27 + CHIR99021 + BMP4. Add Wnt3a if enhanced efficiency is desired. NCC markers (SOX10, p75NTR) should be detectable around Day 10. Use gentle enzymatic dissociation for passaging to avoid loss of fragile NCCs.

Schwann Cell Specification and Maturation

Transfer NCCs to PLO/laminin-coated plates. Switch to Schwann induction medium: DMEM/F12 + N2 + B27 + Heregulin-1β, forskolin, IGF-, BDNF, and PDGF-BB. Change medium every 2 days. By Day 30+, observe elongated, bipolar Schwann cell-like morphology. Gradually reduce forskolin to avoid overproliferation.

Quality Control & Characterization

Marker	Description
S100β	Schwann cell marker
p75NTR	Neural crest & immature Schwann cell
SOX10	Neural crest lineage marker
MBP	Myelin basic protein (mature cells)
GFAP	Intermediate filament protein

Troubleshooting and Optimization Tips

Efficient and reproducible generation of Schwann cells from iPSCs requires precise control of culture conditions, timing of factor exposure, and regular monitoring of cell health. Below is an expanded troubleshooting guide to help resolve common challenges and optimize outcomes across each stage of the protocol.

Problem	Possible Cause	Solution
Low efficiency of neural induction	Inactive CHIR99021 or SB431542 Poor iPSC quality Over-confluent colonies at induction start	Confirm activity of small molecules (freshly prepared or properly stored) Use healthy, undifferentiated iPSCs at ~70% confluency Avoid mechanical disruption during passaging
Neural crest markers not detected	Suboptimal BMP4/Wnt3a dosing Timing deviations in neural crest induction phase	Fine-tune BMP4 concentration Ensure accurate timing Supplement Wnt3a to enhance transition
Excessive cell death after neural crest induction	Incomplete substrate coating Aggressive passaging Serum starvation stress	Use freshly coated PLO/laminin plates Apply gentle dissociation reagents Temporarily include FBS if needed during transition
Poor Schwann cell morphology	Inadequate NRG1 or forskolin signaling Culture too confluent or sparse	Increase NRG1 and verify lot activity Seed at optimal density Refresh medium frequently to maintain growth factor availability
Low expression of S100β or MBP	Incomplete maturation Short induction period Batch variability of supplements	Extend Schwann maturation to 40–50 days if needed Add BDNF or IGF-1 to support maturation Use high-quality, defined supplements to reduce batch effects
Poor cell attachment after plating	Insufficient coating or old matrix Over-trypsinization	Always use freshly prepared PLO/laminin coatings Minimize enzymatic exposure and neutralize quickly
Contamination or morphological abnormalities	Poor aseptic techniques Long-term culture without splitting	Ensure sterile workflow at all stages Do not exceed recommended confluency (>90%) before passaging

Expert Tips from Creative Biolabs

Cell line selection matters: Different iPSC lines have variable neural crest potential—opt for lines with proven ectodermal competence.
Use reporter lines (optional): iPSC lines engineered with SOX10 or S100β-GFP reporters can facilitate real-time monitoring of lineage progression.
Document batch variations: Keep detailed records of media batches, passage numbers, and timing to identify subtle variables affecting results.

Related Services at Creative Biolabs

Creative Biolabs provides a comprehensive portfolio of customized services to support the entire workflow of Schwann cell generation from iPSCs.

Why Choose Creative Biolabs?

Over 20 years of stem cell and neurobiology expertise
End-to-end support from iPSC sourcing to Schwann cell validation
Defined, scalable, and reproducible protocols
Rapid turnaround and customizable deliverables
Dedicated technical support and collaborative flexibility

Creative Biolabs is your ideal partner, providing custom iPSC services and functional validation platforms to support every step of your research journey.

Learn more or request a custom quote.

References

Yoshioka, Yuki, et al. "AAV-mediated editing of PMP22 rescues Charcot-Marie-Tooth disease type 1A features in patient-derived iPS Schwann cells." Communications Medicine 3.1 (2023): 170. https://doi.org/10.1038/s43856-023-00400-y
Distributed under Open Access license CC BY 4.0, without modification.

Created July 2025

For Research Use Only. Not For Clinical Use.