MC3T3-E1 Cells
















General information
Description | MC3T3-E1 is a pre-osteoblastic cell line derived from the calvaria of a mouse embryo. These cells are extensively utilized in the study of osteogenesis, particularly for examining the molecular and cellular mechanisms underlying bone formation and differentiation. The MC3T3-E1 cell line is known for its robust ability to differentiate into osteoblasts in vitro, a process that can be stimulated by ascorbic acid and beta-glycerophosphate. This differentiation is marked by the expression of key osteogenic markers such as alkaline phosphatase, osteocalcin, and type I collagen. MC3T3-E1 cells are instrumental in research focused on bone biology, including the study of bone matrix deposition and mineralization. These cells provide a reliable model for investigating the effects of various drugs, hormones, and genetic modifications on osteoblast function and bone formation. Additionally, the MC3T3-E1 cell line is valuable in studying pathological conditions such as osteoporosis and other bone-related diseases. Their ease of culture and well-characterized response to osteogenic stimuli make them a preferred choice for researchers aiming to unravel the complexities of bone physiology and pathology. |
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Organism | Mouse |
Tissue | Bone, calvaria |
Applications | In vitro osteoblast differentiation |
Synonyms | Mc3T3-E1, MC3T3E1, MC-3T3-E1, MC 3T3-E1 |
Characteristics
Age | 1 day |
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Gender | Unspecified |
Morphology | Fibroblast-like |
Cell type | Osteoblast |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | MC3T3-E1 (Cytion catalog number 305187) |
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Biosafety level | 1 |
Expression / Mutation
Tumorigenic | Yes, in immunodeficient mice |
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Products | Collagen |
Handling
Culture Medium | Alpha MEM, w: 2.0 mM stable Glutamine, w: Ribonucleosides, w: Deoxyribonucleosides, w: 1.0 mM Sodium pyruvate, w: 2.2g/L NaHCO3, w/o: Ascorbic acid (GIBCO, Catalog No. A1049001. We do not supply this product; please consider other suppliers. Please let us know if you need further assistance.) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Doubling time | 24 to 48 hours |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | 1:2 to 1:4 |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) |
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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Required products
- 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.