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MC3T3-E1 Subclone 30
MC3T3-E1 Subclone 30
規格:
貨期:
編號:B165101
品牌:Mingzhoubio

標準菌株
定量菌液
DNA
RNA

規格:
凍干粉
斜面
甘油
平板


產品名稱
MC3T3-E1 Subclone 30
商品貨號
B165101
Organism
Mus musculus, mouse
Tissue
bone, calvaria
Cell Type
preosteoblast
Product Format
frozen
Morphology
fibroblast
Culture Properties
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.

Age
newborn
Strain
C57BL/6
Applications
These cell lines are good models for studying in vitro osteoblast differentiation, particularly ECM signaling.
Storage Conditions
liquid nitrogen vapor phase
Derivation
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.
Genes Expressed
collagen
 RefWang 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
Cellular Products
collagen
Comments
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. 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. 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. After implantation into immunodeficient mice, highly differentiating subclones form bone-like ossicles resembling woven bone, while poorly differentiating cells only produce fibrous tissue. RefWang 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
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%.
Subculturing
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-053mM 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: 1:4 to 1:10
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation
Culture medium 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
Name of Depositor
RT Franceschi
Deposited As
mouse
References

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

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