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MC3T3-E1 Subclone 14 cells are a valuable resource in biological science, specifically in the study of osteoblasts. Derived from a C57BL/6 mouse calvaria, these cells were carefully selected based on their high alkaline phosphatase (ALP) activity while resting.
This unique characteristic makes them an ideal model for investigating osteoblast differentiation and the formation of calcified bone tissue in vitro. As a preosteoblast cell type, MC3T3-E1 Subclone 14 cells exhibit a fibroblast morphology and are primarily associated with bone tissue derived from the calvaria.
One of the notable features of MC3T3-E1 Subclone 14 cells is their ability to differentiate into osteoblasts and osteocytes. Through their extensive morphological and functional resemblance to primary calvarial osteoblasts, these cells offer a reliable platform for studying the extracellular matrix (ECM) signalling and behaviour associated with osteoblast differentiation.
When cultured with ascorbic acid and inorganic phosphate at optimal concentrations (3 to 4 mM), MC3T3-E1 Subclone 14 cells exhibit remarkable levels of osteoblast differentiation. After just ten days, they form a well-mineralized ECM, providing researchers with a window into the intricate process of bone tissue formation.
Moreover, these cells have been found to secrete collagen, an essential component of bone tissue, and express murine leukaemia inhibitory factor (MIF) in RNA. Such characteristics further contribute to their relevance in investigating various biological processes related to bone development and homeostasis. The MC3T3-E1 Subclone 14 cell line has also been employed in cutting-edge research.
For instance, it has been utilized to propose an actin filament cytoskeleton analysis framework, offering insights into the complex intracellular architecture of osteoblasts. Additionally, researchers have explored the effects of biodegradable magnesium and magnesium alloys on these cells, studying their interactions with different materials and their impact on selected cellular properties.
With their diverse applications, these cells are invaluable in 3D cell culture studies, providing a realistic in vitro model for investigating osteoblast behaviour and differentiation within a three-dimensional environment. Their relevance extends to various research fields, including tissue engineering, bone regeneration, and the development of therapeutic interventions for bone-related disorders.
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- Protein expression: Collagen
- Tumorigenic: Yes
- cultureMedium: 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.)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- 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.
- fluidRenewal: 2 to 3 times per week
- freezeMedium: 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.
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