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品系：C57BL/6
組織來源：calvaria
細胞形態(tài)：成纖維樣
細胞類型：其他細胞類型
運輸方式：凍存運輸
是否是腫瘤細胞：0
物種來源：小鼠
ATCC Number：CRL-2593?
器官來源：骨
生長狀態(tài)：貼壁生長
數(shù)量：大量
年限：newborn
規(guī)格：0.1ml Designations: MC3T3-E1 Subclone 4
Depositors: ?RT Franceschi
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Mus musculus
Morphology:fibroblast


Source: Organ: bone
Strain: C57BL/6
Tissue: calvaria
Cell Type: preosteoblast;
Cellular Products:collagen [51540 ]
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Applications:These cell lines are good models for studying in vitro osteoblast differentiation, particularly ECM signaling. They have behavior similar to primary calvarial osteoblasts.
The MC3T3-E1 Subclone 4 (ATCC CRL-2593 ) and the MC3T3 Subclone 14 (ATCC CRL-2594 ) lines exhibit high levels of osteoblast differentiation after growth in ascorbic acid and 3 to 4 mM inorganic phosphate.
Tumorigenic:Yes
Age: newborn
Comments:A series of subclones were isolated from the cloned but phenotypically heterogeneous MC3T3-E1 cell line. The subclones were selected for high or low osteoblast differentiation and mineralization after growth in medium containing ascorbic acid. The MC3T3-E1 Subclone 4 (ATCC CRL-2593 ) and the MC3T3 Subclone 14 (ATCC CRL-2594 ) lines exhibit high levels of osteoblast differentiation after growth in ascorbic acid and 3 to 4 mM inorganic phosphate. They form a well mineralized extracellular matrix (ECM) after 10 days [PubMed: 10352097].
The MC3T3 Subclone 24 (ATCC CRL-2595 ) and the MC3T3 Subclone 30 (ATCC CRL-2596 ) lines exhibit poor osteoblast differentiation after growth in ascorbic acid. They do not form ECM. They can be used as negative controls for Subclones 4 and 14 [PubMed: 10352097].
Mineralizing subclones selectively express mRNAs for the osteoblast markers, bone sialoprotein (BSP), osteocalcin (OCN), and the parathyroid hormone (PTH)/parathyroid hormone-related protein (PTHrP) receptor. Subclones with both high and low differentiation potential produce similar amounts of collagen in culture and express comparable basal levels of mRNA encoding Osf2/Cbfa1, an osteoblast-related transcription factor [PubMed: 10352097].
After implantation into immunodeficient mice, highly differentiating subclones form bone-like ossicles resembling woven bone, while poorly differentiating cells only produce fibrous tissue [PubMed: 10352097].
These cell lines are good models for studying in vitro osteoblast differentiation, particularly ECM signaling. They have behavior similar to primary calvarial osteoblasts.
Propagation: ATCC complete growth medium: The base medium for this cell line is Alpha Minimum Essential Medium with ribonucleosides, deoxyribonucleosides, 2 mM L-glutamine and 1 mM sodium pyruvate, but without ascorbic acid (GIBCO, Custom Product, Catalog No. A1049001). To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol:

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitogen vapor phase
Doubling Time: approximately 38 hours
Related Products:recommended serum:ATCC 30-2020
derived from same cell line:ATCC CRL-2594
derived from same cell line:ATCC CRL-2595
derived from same cell line:ATCC CRL-2596
0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
References: 51540: Wang D, et al. Isolation and characterization of MC3T3-E1 preosteoblast subclones with distinct in vitro and in vivo differentiation/mineralization potential. J. Bone Miner. Res. 14: 893-903, 1999. PubMed: 10352097