HaCaT Cells












Essential facts about Hacat cells
Description | HaCaT cells are a pivotal model in dermatological research, offering insights into the complex mechanisms of skin biology and pathology. The spontaneously immortalized HaCaT cell line is derived from adult human epidermal cells and retains the capacity to proliferate and undergo differentiation, similar to basal keratinocytes in vivo. HaCaT cells serve as a robust platform for investigating the epidermal differentiation process and studying the epidermal differentiation markers essential for maintaining skin integrity. The susceptibility of HaCaT cells to apoptosis and their sensitivity to apoptosis-inducing agents are extensively studied, particularly in the context of cytotoxic agents like RIPL. Researchers assess these agents' cytotoxicities and the extent of cytotoxicity using HaCaT cells, utilizing techniques such as fluorescence microscopy to visualize cellular changes. Researchers have leveraged HaCaT cells to examine the effects of various agents, including antimicrobial substrates and their influence on cell viability. These cells are an excellent substrate for testing antimicrobial biomaterials and antimicrobial atelocollagen substrates, crucial for skin repair and medical applications. The HaCaT epidermal line also plays a crucial role in studying cellular senescence, cytokines, and gene expression profiles related to aging and chronic diseases. The transcriptional profiles of HaCaT cells, including the role of κB and microRNAs, provide insight into the regulatory mechanisms at the molecular level. The HaCaT keratinocyte line, with their characteristics as epidermal keratinocytes, offers a tractable system for dissecting the intricate interplay between epidermal cells and the immune system, specifically the role of keratinocytes in disease states. They enable the exploration of epigenetic modifications and their influence on the differentiation of keratinocytes, including the formation of the cornified envelope, a key feature in the skin's barrier function. In summary, HaCaT cells are an indispensable model in dermatological research, facilitating a deeper understanding of skin biology and pathology through their resemblance to basal keratinocytes and their ability to undergo cell growth and differentiation. Their application spans from studying epidermal differentiation and antimicrobial effects to exploring cellular responses such as apoptosis, making them a cornerstone in cell biology and biomedical research. |
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Organism | Human |
Tissue | Skin |
Details
Age | 62 years |
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Gender | Male |
Ethnicity | Caucasian |
Cell type | Keratinocytes with a diameter of 20-25 micrometer. |
Growth properties | Adherent |
Documentation
Citation | HaCaT (Cytion catalog number 300493) |
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Biosafety level | 1 |
Genetics of the HaCaT keratinocyte cell line
Tumorigenic | No |
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Karyotype | Aneuploid (hypotetraploid) |
Handling the Hacat cell line
Culture Medium | DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | The 1:1 mixture of EDTA (stock. 0.05%) and trypsin (stock: 0.1%) must be prepared each time ahead of detaching the cells using PBS without Ca2+ and Mg2+ to provide a physiologic osmolarity. Ready-to-use mixtures of trypsin/EDTA are not recommended, as this may result in cell clumps. As an alternative, TrypLE Express (Life Technologies) instead of trypsin/EDTA can be used. The protocol of the manufacturer should be followed. |
Doubling time | The doubling time of HaCaT cells is 28 hours. |
Subculturing |
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Split ratio | A ratio of 1:5 to 1:10 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | 2 times per week |
Freeze medium | As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, 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 assurance
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,x
CSF1PO: 9,11
D13S317: 10,12
D16S539: 9,12
D5S818: 12
D7S820: 9,11
TH01: 9.3
TPOX: 11,12
vWA: 16,17
D3S1358: 16
D21S11: 28,30.2
D18S51: 12
Penta E: 7,12
Penta D: 11,13
D8S1179: 14
FGA: 24
D1S1656: 11,12
D2S1338: 17,25
D12S391: 18,23
D19S433: 13,14
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HLA alleles |
A*: '31:01:02
B*: '40:01:02, '51:01:01
C*: '03:04:01, '15:02:01
DRB1*: '04:01:01, '15:01:01
DQA1*: '01:02:01, '03:03:01
DQB1*: '03:01:01, '06:02:01
DPB1*: '03:01:01, '04:01:01
E: '01:03:01, '01:03:02
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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