MC3T3-E1 Cell Line

Brief Introdution
Product List

General Information
Organism	Mus musculus, mouse
Cell Line Description	The osteoblastic cell line MC3T3-E1 was established from C57BL/6 mouse calvaria and selected based on high alkaline phosphatase (ALP) activity at rest. The cells can differentiate into osteoblasts and osteocytes and have been shown to form calcified bone tissue in vitro. The mineral deposit has been identified as hydroxyapatite. Basic fibroblast growth factor (bFGF) mRNA and protein expression is modulated by TGF beta and bFGF treatment. Prostaglandin F2a has been reported to stimulate DNA synthesis and proliferation through upregulation of insulin-like growth factor I receptors. MC3T3-E1 secretes collagen and expresses murine leukemia inhibitory factor (mLIF) in RNA. Contact inhibition is a natural process that prevents cell growth when cells come into contact with each other.
Tissue	Calvaria
Morphology	Fibroblast-like
Strain	C57BL/6
Age	Neonate
Product Format	Frozen
Growth Mode	Adherent
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Applications	1. Bone metabolism research 2. Bone tissue engineering 3. Drug development 4. Cell-cell interaction studies 5. Genetic studies
Shipped in	Dry ice
Storage Temperature	−196°C
Additional Info	It cannot be determined how the loss of contact inhibition affects other properties of the MC3T3-E1 cell line. This cell line catalog number remains a popular choice. A key consideration is whether the intended use of the cell line requires contact inhibitory properties. For example, it is required when testing the effects of potential oncogene expression (where induction of loss of contact inhibition is being measured), whereas it is less relevant when using cell lines expressing recombinant proteins for production and purification.
Characteristics
Tumorigenic	Yes, in immunodeficient mice (forms bone-like ossicles)
Viruses	ELISA: reverse transcriptase negative; PCR: SMRV -
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Remove and discard the culture medium. 2. Briefly rinse the cell layer with a 0.25% (w/v) trypsin-0.53 mM EDTA solution to remove all traces of serum containing trypsin inhibitors. 3. Add 2.0 to 3.0 mL of trypsin-EDTA solution to the flask and observe the cells under an inverted microscope until the cell layer is dispersed (usually within 5 to 15 minutes). NOTE: To avoid clumping, do not agitate the cells by knocking or shaking the flask while waiting for cells to detach. Cells that are difficult to separate can be placed at 37°C to facilitate dispersion. 4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gentle pipetting. 5. Add appropriate aliquots of cell suspension to new culture vessels. 6. Incubate cultures at 37°C. NOTE: The volumes used in this protocol are for a 75 cm² flask. For other sizes of culture vessels, proportionally reduce or increase the amount of dissociation medium.
Medium Renewal	Every 2 to 3 days
Subcultivation Ratio	The ratio of 1:3 to 1:5 is recommended.
Culture Medium	MEM alpha + 2mM Glutamine + 10% Foetal Bovine Serum (FBS)
Culture Conditions	Atmosphere: Air, 95%; CO₂, 5%;Temperature: 37°C
Cryopreservation	Frozen with 70% medium, 20% FBS, 10% DMSO

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* For research use only. Not intended for any clinical use.

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