SH-SY5Y Cells
















Key facts about the SH-SY5Y cell line
Description | SH-SY5Y cells, a subclone derived from the neuroblastoma cancer cell line SK-N-SH, are a valuable cell model for neurodegenerative disorders such as Parkinson's and Alzheimer's disease. The SK-N-SH cell line was established in 1970 from a biopsy of a metastatic bone tumor from a 4-year-old cancer patient. The human SH-SY5Y cell line offers a unique cell source for functional studies in neurobiology and neurodegenerative disease research. SH-SY5Y cells grow both adherently and in suspension, forming clusters during division that differ significantly from the morphology of differentiated cells. These undifferentiated cells, before undergoing neuronal differentiation, serve as an essential foundation for neuroscientific studies. The neuronal differentiation of SH-SY5Y cells, which transforms them into neuronal cell models resembling various functional neurons, is achieved through biochemical interconversion processes involving gradual serum deprivation, retinoic acid, neurotrophic factors like brain-derived neurotrophic factor, and extracellular matrix proteins. This differentiation is crucial for studying neuronal markers and conducting neurotoxicology research, especially concerning the impact of organic pollutants on human neuron-like cells. The neurobiology of SH-SY5Y neuroblastoma cells, primarily known for their dopaminergic characteristics, can be explored for cholinergic properties under specific differentiation conditions. While these cells may express acetylcholinesterase, indicative of some cholinergic activity, their utility in studying cholinergic neurotransmission is less pronounced compared to their role in dopaminergic system studies. As a neurotoxicological model, the SH-SY5Y neuroblastoma cell line is instrumental in examining the effects of compounds on acetylcholinesterase and butyryl cholinesterase activities, essential for neurotoxicology studies. The sy5y line's contribution to understanding the biochemical pathways involved in neurodegenerative diseases, coupled with its role in the functional studies of dopaminergic and cholinergic systems, underscores its value in neuroscience research. |
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Organism | Human |
Tissue | Bone Marrow |
Disease | Neuroblastoma |
Metastatic site | Bone marrow |
Synonyms | SH-Sy5y, SHSY5Y, SHSY-5Y, SK-SH-SY5Y, SY5Y, SH-SY5Y Parental |
Specifications
Age | 4 years |
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Gender | Female |
Morphology | The cells grow as clusters of neuroblastic cells with multiple, short, fine cell processes (neurites). Cells will aggregate, form clumps and float. A confluent monolayer is not formed. |
Cell type | Neuroblast |
Growth properties | Loosely adherent and form clumps at high cell density |
Documentation
Citation | SH-SY5Y (Cytion catalog number 300154) |
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Biosafety level | 1 |
Genetic profile
Tumorigenic | Forms tumors in nude mice within approx. 3-4 weeks. |
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Karyotype | The cytogenetic landscape of SH-SY5Y cells is marked by complex chromosomal aberrations, notably featuring a modal chromosome number of 47, including trisomy of 1q due to a distinctive insertion in chromosome 1. This genetic backdrop is crucial for understanding the cellular biology and oncogenic potential of SH-SY5Y cells, making them a versatile model in neuroscientific research, particularly in the realms of neurodevelopment, neurotoxicity, and neurodegenerative disease studies. |
Handling the SH-SY5Y neuroblastoma cell line
Culture Medium | Please mix EMEM and Ham's F12 in a 50:50 ratio (Cytion article numbers 820100a and 820600a) |
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Medium supplements | Supplement the medium with 15% FBS |
Passaging solution | Accutase |
Subculturing | These cells grow as a mixture of floating and adherent cells. Remove the medium with the floating cells, and recover the cells by centrifugation. Rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add Accutase (1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at 37 degree Celsius for 10 minutes. Combine with the floating cells recovered above. Carefully resuspend the cells, the addition of medium is optional but not necessary, and dispense into new flasks which contain fresh medium. |
Seeding density | Seeding density after thawing 6x10^4 cells/cm^2, seed into 1x T25 cell culture flask. The cells will become 80-90% confluent within 1-2 weeks. Once the cells proliferate vigorously, seed out the cells at a density of 1 - 2 x10^4 cells/cm^2. |
Fluid renewal | 1 to 2 times per week |
Freeze medium | As a cryopreservation medium, use 50% basal medium + 40% FBS + 10% DMSO, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. |
Handling of cryopreserved cultures |
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Quality control
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,y
CSF1PO: 11
D13S317: 11
D16S539: 8,13
D5S818: 12
D7S820: 7,10
TH01: 7,10
TPOX: 8,11
vWA: 14,18
D3S1358: 15,16
D21S11: 31,31.2
D18S51: 13,16
Penta E: 7,11
Penta D: 10,12
D8S1179: 15
FGA: 23.2,24
D6S1043: 12,18
D2S1338: 17,19
D12S391: 18,22
D19S433: 13,14
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HLA alleles |
A*: '01:01:01, '24:02:01
B*: '18:01:01, '49:01:01
C*: '07:01:01
DRB1*: '11:04:01, '13:01:01
DQA1*: '01:03:01, '05:05:01
DQB1*: '03:01:01, '06:03:01
DPB1*: '02:01:02, '04:01:01
E: '01:01, '01:03
|
Required products
What sets DMEM apart from other media is its unique composition. It contains an impressive fourfold increase in amino acid and vitamin concentration compared to the original Eagle's Minimal Essential Medium. Initially developed with low glucose (1 g/L) and sodium pyruvate, DMEM is frequently employed with higher glucose levels, either with or without sodium pyruvate. Notably, DMEM does not contain proteins, lipids, or growth factors, necessitating supplementation. To achieve optimal growth, a common approach is to supplement DMEM with 10% Fetal Bovine Serum (FBS). Additionally, DMEM employs a sodium bicarbonate buffer system, requiring a 5-10% CO2 environment to maintain a physiological pH.
Dulbecco's Modified Eagle Medium (DMEM) is highly regarded among defined media for cell and tissue culture, catering to the growth needs of various adherent cell phenotypes. It surpasses the original Eagle's Medium, developed in the 1950s for cultivating chicken cells, through the enhanced supplementary formulation known as Dulbecco's modification. This modification significantly elevates the content of select amino acids and vitamins up to fourfold compared to the original medium.
In the field of cell culture, DMEM plays a vital role as a growth medium for different cell types, including primary cells, stem cells, and transformed cells. Researchers also employ the modified version of DMEM for a wide array of research applications, such as drug discovery, tissue engineering, and the study of cell signaling pathways.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +8°C, protected from light.
Once opened, store at 4°C and use within 6–8 weeks.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Amino Acids
Glycine
30.00
L-Arginine HCl
84.00
L-Cystine 2 HCl
62.57
L-Glutamine
584.00
L-Histidine HCl H2O
42.00
L-Isoleucine
105.00
L-Leucine
105.00
L-Lysine HCl
146.00
L-Methionine
30.00
L-Phenylalanine
66.00
L-Serine
42.00
L-Threonine
95.00
L-Tryptophan
16.00
L-Tyrosine 2 Na 2H2O
103.79
L-Valine
94.00
Vitamins
Choline chloride
4.00
D-Calcium Pantothenate
4.00
Folic Acid
4.00
myo-Inositol
7.20
Nicotinamide
4.00
Pyridoxal HCl
4.00
Riboflavin
0.40
Thiamine HCl
4.00
Inorganic Salts
CaCl2 2 H2O
265.00
Fe(NO3)3 9 H2O
0.10
KCl
400.00
MgSO4 7H2O
200.10
NaCl
6400.00
NaHCO3
3700.00
NaH2PO4 2H2O
141.73
Other Components
D-Glucose
4500.00
Phenol Red Sodium Salt
15.90
Sodium Pyruvate
110.00
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.
Phosphate-buffered saline (PBS) is a widely used buffer solution in biological and chemical research. It plays a crucial role in maintaining the pH balance and osmolarity during various experimental procedures, including tissue processing and cell culture. Our PBS solution is meticulously formulated with high-purity ingredients to ensure stability and reliability in every experiment. The osmolarity and ion concentrations of our PBS closely mimic those of the human body, making it isotonic and non-toxic to most cells.
Composition of Our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains:
8000 mg/L Sodium chloride (NaCl)
200 mg/L Potassium chloride (KCl)
1150 mg/L Sodium phosphate dibasic anhydrous (Na2HPO4)
200 mg/L Potassium phosphate monobasic anhydrous (KH2PO4)
This composition ensures an optimal pH and ionic balance, suitable for a wide range of biological applications.
Applications of Our PBS Solution
Our PBS solution is ideal for various applications in biological research. Its isotonic and non-toxic properties make it suitable for substance dilution and cell container rinsing. PBS solutions containing EDTA are effective for disengaging attached and clumped cells. However, divalent metals such as zinc should not be added to PBS, as this can cause precipitation. In such cases, Good's buffers are recommended. Additionally, our PBS solution is an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, including SARS-CoV-2.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +25°C, protected from light.
Once opened, store at 2°C to 25°C and use within 24 months.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Salts
Potassium chloride
200
Potassium phosphate monobasic anhydrous
200
Sodium chloride
8000
Sodium phosphate dibasic anhydrous
1150