SH-SY5Y Cell Line

SH-SY5Y Cell Line

Overview

SH-SY5Y Cell Line

Overview

<table class="infobox infobox-cell">
<tr>
<th class="infobox-header" colspan="2">SH-SY5Y Cell Line</th>
</tr>
<tr>
<td class="label">Species</td>
<td>Human (Homo sapiens)</td>
</tr>
<tr>
<td class="label">Tissue</td>
<td>Bone marrow (metastatic neuroblastoma)</td>
</tr>
<tr>
<td class="label">Age</td>
<td>4 years old (donor)</td>
</tr>
<tr>
<td class="label">Sex</td>
<td>Female</td>
</tr>
<tr>
<td class="label">Karyotype</td>
<td>Modal number = 47 (trisomy 1q)</td>
</tr>
<tr>
<td class="label">Blood type</td>
<td>A, Rh+</td>
</tr>
<tr>
<td class="label">Growth pattern</td>
<td>Mixed adherent/suspension</td>
</tr>
<tr>
<td class="label">Population doubling time</td>
<td>~48 hours</td>
</tr>
<tr>
<td class="label">ATCC Catalog</td>
<td>CRL-2266</td>
</tr>
<tr>
<td class="label">Agent</td>
<td>Concentration</td>
</tr>
<tr>
<td class="label">dbcAMP</td>
<td>1 mM</td>
</tr>
<tr>
<td class="label">GDNF</td>
<td>20 ng/mL</td>
</tr>
<tr>
<td class="label">TPA</td>
<td>80 nM</td>
</tr>
<tr>
<td class="label">DAPT</td>
<td>10 muM</td>
</tr>
<tr>
<td class="label">Cell Line</td>
<td>Origin</td>
</tr>
<tr>
<td class="label">SK-N-SH</td>
<td>Neuroblastoma (parent line)</td>
</tr>
<tr>
<td class="label">SK-N-Be(2)</td>
<td>Neuroblastoma</td>
</tr>
<tr>
<td class="label">PC12</td>
<td>Rat pheochromocytoma</td>
</tr>
<tr>
<td class="label">LUHMES</td>
<td>Immortalized human neurons</td>
</tr>
<tr>
<td class="label">iPSC-derived neurons</td>
<td>Induced pluripotent stem cells</td>
</tr>
</table>

SH-SY5Y is a human neuroblastoma cell line widely used as an in vitro model for studying neurodegenerative diseases, particularly Parkinson's disease (PD), Alzheimer's disease (AD), and related disorders. This cell line serves as a valuable tool for investigating neuronal differentiation, synaptic function, neurotoxicity, and therapeutic drug screening.

Origin and Derivation

SH-SY5Y is a subclone of the SK-N-SH cell line, which was originally established in 1970 from a metastatic bone tumor of a 4-year-old female patient with neuroblastoma[@biedler1978]. The SK-N-SH line was subsequently cloned to generate the SH-SY5Y subclone, which exhibits enhanced neuronal differentiation potential compared to the parent line[@atcc].

Key Characteristics

Biology and Phenotype

Undifferentiated State

In their undifferentiated state, SH-SY5Y cells exhibit a neuroblast-like morphology with both adherent and non-adherent populations. They express low levels of neuronal markers and retain some characteristics of proliferative neural crest-derived cells. Key features include:

• Neuronal markers: Low expression of tyrosine hydroxylase (TH), dopamine transporter (DAT), and synaptic proteins
• Neurotrophic receptors: Express NGF receptor (TrkA) and GDNF receptor (GFRα1)
• Metabolic activity: Active mitochondria, capable of oxidative phosphorylation
• Signal transduction: Functional PI3K/Akt, MAPK/ERK, and Wnt/β-catenin pathways

Differentiated State

Upon treatment with differentiation agents, SH-SY5Y cells undergo morphological and biochemical changes resembling mature [neurons](/entities/neurons):

• Neurite outgrowth: Extension of long, branching neurites
• Synaptic formation: Expression of synaptophysin, synapsin, and PSD95
• Neurotransmitter synthesis: Increased TH and aromatic L-amino acid decarboxylase (AADC) activity
• Electrophysiology: Development of voltage-gated ion channels and action potentials

Differentiation Protocols

Retinoic Acid (RA) Differentiation

Retinoic acid is the most commonly used differentiation agent for SH-SY5Y cells. RA activates nuclear retinoic acid receptors (RARs), which regulate gene expression programs involved in neuronal maturation[@encinas2000].

Protocol Overview:

Molecular Mechanisms:

• Upregulation of neuronal differentiation genes (NeuroD1, Ngn2, MAP2)
• Increased expression of TH, DAT, and VMAT2
• Activation of PI3K/Akt and MAPK/ERK signaling

BDNF-Induced Differentiation

Brain-derived neurotrophic factor (BDNF) promotes neuronal survival and differentiation through TrkB receptor activation[@kaplan1991]. BDNF treatment following RA priming produces more mature dopaminergic neurons.

Protocol Overview:

Alternative Protocols

Applications in Parkinson's Disease Research

SH-SY5Y cells are extensively used as a Parkinson's disease model due to their ability to differentiate into dopaminergic-like neurons. Key research applications include:

α-Synuclein Pathology

• Overexpression models: Wild-type and mutant [α-synuclein](/proteins/alpha-synuclein) (A30P, A53T) transfection to study aggregation
• Toxicity studies: Proteasomal inhibition, oxidative stress, and mitochondrial dysfunction
• Lewy body formation: Visualization of insoluble aggregates

Mitochondrial Dysfunction

• Complex I inhibition: Rotenone and MPTP treatment to model PD
• Oxidative stress: Hydrogen peroxide, 6-OHDA exposure
• Mitophagy studies: Parkin and PINK1 knockdown/overexpression

Neuroprotective Drug Screening

SH-SY5Y cells serve as a high-throughput screening platform for potential PD therapeutics:

• Antioxidants: CoQ10, vitamin E, NAC
• [Autophagy](/entities/autophagy) enhancers: Rapamycin, trehalose
• Anti-apoptotic compounds: Bcl-2 family modulators
• Iron chelators: Deferoxamine, clioquinol

LRRK2 Studies

[LRRK2](/entities/lrrk2) (leucine-rich repeat kinase 2) is the most common genetic cause of familial PD. SH-SY5Y cells with LRRK2 mutations (G2019S, R1441C/G/H) are used to study:

• Kinase activity modulation
• Neuronal viability effects
• Synaptic function impairment

Applications in Alzheimer's Disease Research

Beyond PD, SH-SY5Y cells are valuable for AD research:

Amyloid-β Toxicity

• Aβ1-42 peptide treatment to model amyloid toxicity
• [APP](/entities/app-protein) processing and amyloidogenic cleavage studies
• Synaptic dysfunction mechanisms

Tau Pathology

• [Tau](/proteins/tau) overexpression and phosphorylation studies
• NFT formation models
• Microtubule stability investigations

Neurotrophin Signaling

• NGF and BDNF signaling in neuronal survival
• Trk receptor activation studies
• Neuroprotective pathway elucidation

Advantages and Limitations

Advantages

• Human origin: More relevant than rodent models for human disease
• Dopaminergic potential: Can be differentiated into TH-positive neurons
• Ease of culture: Relatively straightforward maintenance and manipulation
• Genetic manipulability: Transfection, transduction, and CRISPR editing feasible
• Cost-effective: Less expensive than primary neurons or iPSC-derived models

Limitations

• Tumor-derived: Retains some proliferative capacity even when differentiated
• Genetic instability: Variable karyotype across passages
• Limited maturation: Differentiated cells not fully equivalent to primary neurons
• Species-specific: Findings may not fully translate to human physiology

Related Cell Lines

See Also

• [Alzheimer's Disease](/diseases/alzheimers-disease)
• [Parkinson's Disease](/diseases/parkinsons-disease)

External Links

• [PubMed](https://pubmed.ncbi.nlm.nih.gov/)
• [KEGG Pathways](https://www.genome.jp/kegg/pathway.html)

Pathway Diagram

The following diagram shows the key molecular relationships involving SH-SY5Y Cell Line discovered through SciDEX knowledge graph analysis: