PC-12 Cells
























General information
Description | PC-12 cells are a cell line derived from a pheochromocytoma of the rat adrenal medulla. These cells are of embryonic origin, grow adherently and resemble a mixture of neuroblastic and eosinophilic cells. PC-12 cells are catecholamine cells that synthesize, store and release norepinephrine and dopamine. They have a diameter of approximately 10-12 microns and are small, irregularly shaped cells. The PC12 cell line is a classical neuronal cell model due to its ability to acquire sympathetic neuron features when dealing with nerve growth factor (NGF). Studies on dopamine regulation have shown that PC12 cells synthesize, release, and reuptake dopamine and have been extensively characterized for neurosecretion and the presence of ion channels and neurotransmitter receptors. Moreover, the relative proportion of various subtypes of Ca channels changes during differentiation. The PC12 cell line is an established neuronal cell model that is particularly useful in studying cellular responses to nerve growth factors (NGF) and how these lead to the expression of differentiation-specific proteins and differentiation. When cultured in NGF, PC12 cells differentiate into sympathetic ganglion neurons morphologically and functionally. The differentiation results from the reversible induction of a neuronal phenotype by NGF. Collagen coating has been shown to be favourable to achieving neuronal characteristics in terms of length and density of neurites by NGF treatment. PC12 cells are tumorigenic and were derived from male New England Deaconess Hospital strain rats. The PC-12 cell line has 40 chromosomes, 38 autosomes, plus xY. Nerve growth factor (NGF) is expressed in PC12 cells, and exposure to NGF is one crucial regulator of cell differentiation. In conclusion, PC12 cells are a versatile and widely used model system in neurobiology due to their ability to acquire sympathetic neuron features when dealing with nerve growth factor (NGF). These cells have been extensively characterized for neurosecretion, ion channels, and neurotransmitter receptors. Their extreme versatility for pharmacological testing and use as an established model for studying the proliferation and differentiation of neuronal cells make them a valuable tool in neurobiology research. |
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Organism | Rat |
Tissue | Adrenal gland |
Disease | Pheochromocytoma |
Synonyms | PC 12, PC12 |
Characteristics
Age | Unspecified |
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Gender | Male |
Ethnicity | Japanese |
Morphology | Polygonal |
Growth properties | Small clusters in suspension, poorly adherent, patches on collagen. |
Identifiers / Biosafety / Citation
Citation | PC-12 (Cytion catalog number 500311) |
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Biosafety level | 1 |
Expression / Mutation
Receptors expressed | Nerve growth factor (NGF) |
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Tumorigenic | Yes, in New England Deaconess Hospital strain rats |
Products | Catecholamines, dopamine |
Karyotype | 40 chromosomes, 38 autosomes plus xY |
Handling
Culture Medium | RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) |
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Medium supplements | Supplement the medium with 10% FBS |
Subculturing | Suspension cells: Remove cells from substrate by pipetting with fresh medium. To obtain single cells, pass the suspension several times through a 22 gauge needle and dispense into new flasks. Growing on collagen: To remove adherent cells, use the following standard protocol. Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add TrypleExpress (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. Carefully resuspend the cells, the addition of medium is optional but not necessary, and dispense into new flasks which contain fresh medium. |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 48 hours. |
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 / Genetic profile / HLA
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 |
Rat_D1Wox31: 100
Rat_D2Wox37: 156
Rat_D19Wox11: 228
Rat_D10Wox8: 262,266
Rat_D4Wox7: 145
Rat_D2Wox27: 207
Rat_D5Rat33: 116,118,120
Rat_D10Wox11: 174
Rat_D1Wox23: 226,230
Rat_D12Wox1: 402,406
Rat_D6Wox2: 104
Rat_D8Wox7: 182
Rat_D6Cebr1: 229,231,233
SRY: x,Y
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Required products
Initially designed to support the growth of human leukemic cells in both suspension and monolayer cultures, RPMI 1640 Medium has evolved through modifications by researchers and commercial suppliers to become suitable for a diverse range of mammalian cells. It is exceptionally compatible with cell lines such as HeLa, Jurkat, MCF-7, PC12, PBMC, astrocytes, and carcinomas.
RPMI 1640 Medium stands apart from other cell culture media due to its unique composition. It contains a substantial amount of phosphate, amino acids, and vitamins. Notably, it encompasses biotin, vitamin B12, and PABA, absent in Eagle's Minimal Essential Medium or Dulbecco's Modified Eagle Medium. Moreover, RPMI 1640 Medium exhibits significantly elevated concentrations of vitamins inositol and choline. However, it does not contain proteins, lipids, or growth factors. Consequently, supplementation with 10% Fetal Bovine Serum (FBS) is commonly required to provide optimal conditions for cell growth.
The buffering system of RPMI 1640 Medium relies on sodium bicarbonate and necessitates a 5-10% CO2 environment to maintain a physiologically appropriate pH. The inclusion of the reducing agent glutathione further distinguishes this medium from others.
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
10.00
L-Alanyl-L-Glutamine
434.40
L-Arginine
200.00
L-Asparagine H2O
56.82
L-Aspartic Acid
20.00
L-Cystine 2HCl
65.20
L-Glutamic Acid
20.00
L-Histidine HCl H2O
20.27
L-Hydroxy-L-Proline
20.00
L-Isoleucine
50.00
L-Leucine
50.00
L-Lysine HCl
40.00
L-Methionine
15.00
L-Phenylalanine
15.00
L-Proline
20.00
L-Serine
30.00
L-Threonine
20.00
L-Tryptophan
5.00
L-Tyrosine 2Na 2H2O
28.83
L-Valine
20.00
Vitamins
p-Amino Benzoic Acid
1.00
D-Biotin
0.20
Choline Chloride
3.00
D-Calcium Pantothenate
0.25
Folic Acid
1.00
myo-Inositol
35.00
Nicotinamide
1.00
Pyridoxine HCl
1.00
Riboflavin
0.20
Thiamine HCl
1.00
Vitamin B12
0.005
Inorganic Salts
Ca(NO3)2 4H2O
100.00
KCl
400.00
MgSO4 7H2O
100.00
NaCl
6000.00
NaHCO3
2000.00
Na2HPO4
800.00
Other Components
D-Glucose
2000.00
L-Glutathione Reduced
1.00
Phenol Red Sodium Salt
5.30
- 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