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器官來源：骨
細(xì)胞形態(tài)：成纖維樣
生長狀態(tài)：貼壁生長
數(shù)量：大量
是否是腫瘤細(xì)胞：0
物種來源：小鼠
年限：newborn
運輸方式：凍存運輸
ATCC Number：CRL-2596?
品系：C57BL/6
組織來源：calvaria
規(guī)格：0.1ml Designations: MC3T3-E1 Subclone 30
Depositors: ?RT Franceschi
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Mus musculus deposited as mouse
Morphology:fibroblast


Source: Organ: bone
Strain: C57BL/6
Tissue: calvaria
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.
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.
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. [51540 ]
They form a well mineralized extracellular matrix (ECM) after 10 days.
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. [51540 ]
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. [51540 ]
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. [51540 ]
After implantation into immunodeficient mice, highly differentiating subclones form bone-like ossicles resembling woven bone, while poorly differentiating cells only produce fibrous tissue. [51540 ]
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%.
Temperature: 37.0°C
Subculturing: Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:10 is recommended
Medium Renewal: Every 2 to 3 days
Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37°C) until the cells detach.
Add fresh culture medium, aspirate and dispense into new culture flasks.
Preservation: culture medium 95%; DMSO, 5%
Related Products:recommended serum: ATCC 30-2020
derived from same cell line:ATCC CRL-2593
derived from same cell line:ATCC CRL-2594
derived from same cell line:ATCC CRL-2595
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