Considerations for the use of SH-SY5Y neuroblastoma cells in neurobiology
Open accessAbstract
The use of primary mammalian neurons derived from embryonic central nervous system tissue is limited by the fact that once terminally differentiated into mature neurons, the cells can no longer be propagated. Transformed neuronal-like cell lines can be used in vitro to overcome this limitation. However, several caveats exist when utilizing cells derived from malignant tumors. In this context, the popular SH-SY5Y neuroblastoma cell line and its use in in vitro systems is described. Originally derived from a metastatic bone tumor biopsy, SH-SY5Y (ATCC(®) CRL-2266™) cells are a subline of the parental line SK-N-SH (ATCC(®) HTB-11™). SK-N-SH were subcloned three times; first to SH-SY, then to SH-SY5, and finally to SH-SY5Y. SH-SY5Y were deposited to the ATCC(®) in 1970 by June L. Biedler.Three important characteristics of SH-SY5Y cells should be considered when using these cells in in vitro studies. First, cultures include both adherent and floating cells, both types of which are viable. Few studies address the biological significance of the adherent versus floating phenotypes, but most reported studies utilize adherent populations and discard the floating cells during media changes. Second, early studies by Biedler's group indicated that the parental differentiated SK-N-SH cells contained two morphologically distinct phenotypes: neuroblast-like cells and epithelial-like cells (Ross et al., J Nat Cancer Inst 71:741-747, 1983). These two phenotypes may correspond to the "N" and "S" types described in later studies in SH-SY5Y by Encinas et al. (J Neurochem 75:991-1003, 2000). Cells with neuroblast-like morphology are positive for tyrosine hydroxylase (TH) and dopamine-β-hydroxylase characteristic of catecholaminergic neurons, whereas the epithelial-like counterpart cells lacked these enzymatic activities (Ross et al., J Nat Cancer Inst 71:741-747, 1983). Third, SH-SY5Y cells can be differentiated to a more mature neuron-like phenotype that is characterized by neuronal markers. There are several methods to differentiate SH-SY5Y cells and are mentioned below. Retinoic acid is the most commonly used means for differentiation and will be addressed in detail.
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Differentiated SH-SY5Y neuroblastoma cells fail to replicate mature neuronal morphology, dendritic and axonal complexity, or electrophysiological and synaptic protein expression.
“In contrast, Kovalevich and Langford (2013) reported that differentiated SH-SY5Y neuroblastoma cells failed to replicate mature neuronal morphology, dendritic and axonal complexity, or electrophysiological and synaptic protein expression.”
From Primary cell cultures versus immortalized cell lines: critical comparative insights into biological fidelity and translational applications (2026) · cites this paper as Kovalevich and Langford (2013)
Section 1.2 says differentiated SH-SY5Y cells become more similar to primary neurons, developing long processes that may be numerous or polarized; Section 1.5 also reports neurite branching. Section 1.7 says they express mature neuronal and synaptic proteins, including synaptophysin and SAP-97. The paper does not claim they fully reproduce mature neurons, but its account is contrary to the attributed blanket failure. (LLM, conf=0.90)
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Experimental — accuracy not yet measuredSH-SY5Y cells can have a mixed culture and differentiate into specialized neurons through markers, allowing adrenergic, cholinergic, and dopaminergic models, and are a basic model for neurodegenerative pathologies such as Alzheimer's disease.
“Their most important characteristics are their ability to have a mixed culture (adherent and in suspension) and to differentiate into specialized neurons through markers, allowing adrenergic, cholinergic, and dopaminergic models [142,143], to be a basic model for the study of neurodegenerative pathologies, such as AD.”
From Multi-Target Mechanisms of Phytochemicals in Alzheimer’s Disease: Effects on Oxidative Stress, Neuroinflammation and Protein Aggregation (2022) · cites this paper as [143]
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“In contrast, Kovalevich and Langford (2013) reported that differentiated SH-SY5Y neuroblastoma cells failed to replicate mature neuronal morphology, dendritic and axonal complexity, or electrophysiological and synaptic protein expression.”
- Authors
- Kovalevich, J., & Langford, D.
- Affiliations
- Temple University (US)
- Venue
- Methods in molecular biology
- Publisher
- Humana Press
- Published
- 2013
- Volume
- 1078
- Pages
- 9-21
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- PMC
- PMC5127451
- OpenAlex
- W2401296772
- Cited by
- 1,005
- References
- 39
- Citations by year
- 2026: 84 · 2025: 109 · 2024: 131 · 2023: 119 · 2022: 120 · 2021: 117 · 2020: 102 · 2019: 71 · 2018: 57 · 2017: 37 · 2016: 29 · 2015: 16 · 2014: 10 · 2013: 2
- FWCI
- 52.51
- Type
- book chapter
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of Mental Health (R01MH085602, P30MH092177)
- Topics
- Health Sciences › Medicine › Neurology › Neuroblastoma Research and Treatments
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Keywords
- SH-SY5Y cells, neuroblastoma cell lines, SK-N-SH cells, neuronal differentiation, retinoic acid, catecholaminergic neurons, tyrosine hydroxylase, dopamine beta-hydroxylase, neuron-like cells, in vitro neuronal model
- MeSH terms
- Cell Differentiation, Humans, Neurobiology, Neuroblastoma, Cryopreservation, Cell Culture Techniques, Cell Line, Tumor
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