Human Gingival Fibroblasts (hGF)

Accutase Cell Dissociation Reagent
- 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.

DMEM:Ham's F12 is a widely recognized and extensively utilized basal medium in cell culture for biological research. It serves as a fundamental source of nutrients for the growth of various mammalian cell lines, particularly when supplemented with Fetal Bovine Serum (FBS).

This unique formulation combines Dulbecco's Modified Eagle Medium (DMEM) and Ham's F-12 (Ham's Nutrient Mixture F-12) in a precise 1:1 ratio. The addition of L-glutamine further enhances its composition.

DMEM, derived from Eagle's Minimal Essential Medium (EMEM), offers an increased concentration of amino acids and vitamins compared to its predecessor. In contrast, Ham's F-12 is based on Ham's F-10 medium, providing a complementary set of essential components.

To support optimal cell growth, it is common practice to supplement DMEM:Ham's F12 with FBS at a typical concentration of 5-10%. This addition is necessary as the medium lacks growth hormones, lipids, and proteins crucial for cellular development.

DMEM:Ham's F12 incorporates a pH buffer system and is often supplemented with phenol red, a pH indicator. Cultured cells in DMEM:Ham's F12, or any medium utilizing the bicarbonate buffer system, require a controlled CO2 environment of 5-10% to maintain appropriate pH levels. Phenol red enables monitoring of pH changes from 6.2 (yellow) to 8.2 (red).
Quality Control

pH = 7.2 +/
- 0.02 at 20-25°C.  
Each lot has been tested for sterility and absence of mycoplasma and bacteria. 

Maintenance

Keep refrigerated at +2°C to +8°C in the dark. Freezing as well as warming up to +37°C minimize the quality of the product.  
Do not heat the medium to more than 37°C or use uncontrollable sources of heat (e.g., microwave appliances).  
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature. 
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture. 

Composition




Components
mg/L


Inorganic Salts
Calcium chloride x 2H2O
154,45



Iron(III)-nitrate x 9H2O
0,05



Iron(II)-sulfate x 7H2O
0,42



Potassium chloride
311,83



Copper(II)-sulfate x 5H2O
0.001



Magnesium chloride anhydrous
28,57



Magnesium sulfate
48,85



Sodium chloride
6,999.50



Sodium dihydrogen phosphate anhydrous
54,35



di-Sodium hydrogen phosphate
70,98



Zinc sulfate x 7H2O
0,43


Other Components
D(+)-Glucose anhydrous
3,151.00



HEPES
3,574.50



Hypoxanthine
2,04



Linoleic acid
0,04



DL-68-Lipoic acid
0.103



Sodium pyruvate
110,00



Phenol red
8,10



Putrescin x 2HCl
0.081



Thymidine
0,36


Amino Acids
L-Alanine
4,45



L-Arginine x HCl
147,35



L-Asparagine x H2O
7,50



L-Aspartic acid
6,65



L-Cystine x 2HCl
31,29



L-Cysteine x HCl x H2O
17,56



L-Glutamine
365,00



L-Glutamic acid
7,35



Glycine
18,75



L-Histidine x HCl x H2O
31,48



L-Isoleucine
54,37



L-Leucine
58,96



L-Lysine x HCl
91,37



L-Methionine
17,24



L-Phenylalanine
35,48



L-Proline
17,27



L-Serine
26,25



L-Threonine
53,55



L-Tryptophan
9,02



L-Tyrosine x 2Na x 2H2O
55,81



L-Valine
52,66


Vitamins
D-(+)-Biotine
0,004



D-Calcium pantothenate
2,12



Choline chloride
8,98



Folic acid
2,66



myo-Inositol
12,51



Nicotinamide
2,02



Pyridoxine x HCl
2,03



Riboflavin
0,22



Thiamine x HCl
2,17



Vitamine B12
0,68



NaHCO3
1,200.00