• 名称: RAW 264.7(小鼠单核巨噬细胞白血病细胞)
  • 货号: CBP60533
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CBP60533  

Synonyms:

RAW264; RAW2647; RAW264.7; RAW-264.7; Raw 264.7; Raw264.7

Background:

Established from an ascites of a tumour induced in a male mouse by intraperitoneal injection of Abelson Leukaemia Virus (A-MuLV). Cells will pinocytose neutral red and phagocytose zymosan. Cells capable of antibody dependent lysis of sheep erythrocytes and tumour targets. Growth inhibited by LPS.

Species:

Mus musculus, mouse

Tissue:

Abelson murine leukemia virus-induced tumor; ascites

Disease:

Abelson murine leukemia virus-induced tumor

Gender:

Male adult

Morphology:

Monocyte/macrophage

Growth Mode:

Adherent

Doubling Time:

NA

DNA Profile:

NA
Cobioer’s Cell Line Authentication Service

Culture Medium:

DMEM+10%FBS
We strongly suggest to purchase the complete medium from Cobioer.

Cryopreservation medium:

90% FBS+10%DMSO

Photo:

 

Antigen Expression:

H-2d

Receptor Expression:

Complement (C3)

Applications:

This cell line is a suitable transfection host.

The cells will pinocytose neutral red and will phagocytose latex beads and zymosan. They are capable of antibody dependent lysis of sheep erythrocytes and tumor cell targets. LPS or PPD treatment for 2 days stimulates lysis of erythrocytes but not tumor cell targets.

Comments:

This cell line is easy to propagate, high efficiency for DNA transfection, sensitivity to RNA interference, and supports replication of murine noroviruses. This cell line is negative for surface immunoglobulin (sIg-), Ia (Ia-) and Thy-1.2 (Thy-1.2). When this line was established, it was described as not secreting detectable virus particles and negative using the XC plaque formation assay. Based on a published study by Dr. Janet W. Hartley in 2008, this line was demonstrated to express ecotropic and polytropic MuLV, and is positive using the XC plaque assay for virus replication.

For more information, please contact Cobioer (4008-750-250).

 

II. Handling Procedure for Flask Cultures

The flask was seeded with cells grown and completely filled with medium at Cobioer to prevent loss of cells during shipping.
1.Upon receipt visually examine the culture for macroscopic evidence of any microbial contamination.
Using an inverted microscope (preferably equipped with phasecontrast optics), carefully check for any evidence of microbial contamination. Also check to determine if the majority of cells are still attached to the bottom of the flask; during shipping the cultures are sometimes handled roughly and many of the cells often detach and become suspended in the culture medium (but are still viable).
2.If the cells are still attached, aseptically remove all but 5 to 10 mL of the shipping medium. The shipping medium can be saved for reuse. Incubate the cells at 37°C with 5% CO2 in air atmosphere until they are ready to be subcultured.
3.If the cells are not attached, aseptically remove the entire contents of the flask and centrifuge at 1000rpm for 5 to 10 minutes. Remove shipping medium and save. Resuspend the pelleted cells in 10 mL of this medium and add to 25 cm2 flask. Incubate at 37°C with 5% CO2 in air atmosphere until cells are ready to be subcultured.

 

III. Subculturing Procedure

Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
1.Remove and discard culture medium.
2.Briefly rinse the cell layer with 0.25% (w/v) Trypsin0.53 mM EDTA solution to remove all traces of serum that 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 (depend on the batch of trypsin).
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 with 5% CO2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week

 

IV. Cryopreservation Procedure

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) Trypsin0.53mM 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 1 to 10 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.To remove trypsin-EDTA solution, transfer cell suspension to a centrifuge tube and spin at approximately 1000rpmfor 5 to10 minutes.
6.Discard supernatant and resuspend cells in cryopreservation medium.
7.Transfer the cells into Cryogenic Vials, 1ml/vial.
8.Frozen the cells in cryogenic container (Nalgene #5100-0001).

 

V. Database

Mutation:

Mutation Gene

Zygosity

Gene Sequence

Protein Sequence

NA

NA

NA

NA

For more information, Please visit
http://www.cobioer.com/sjk/&pmcId=77.html

Expression:

NA
For more information, Please visit
http://www.cobioer.com/sjk/&pmcId=77.html

Comments:

For more information, please contact Cobioer(4008-750-250).

 

VI. References

1. PubMed=25504905; DOI=10.1002/pmic.201400431; Guo M., Hartlova A., Dill B.D., Prescott A.R., Gierlinski M., Trost M.; "High-resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow derived macrophages."; Proteomics 15:3169-3174(2015).

2.PubMed: 8662857; Lokuta MA, et al. Mechanisms of murine RANTES chemokine gene induction by newcatle disease virus. J. Biol. Chem. 271: 13731-13738, 1996.

3. PubMed: 8663413; Taylor MF, et al. In vitro efficacy of morpholino-modified antisense oligomers directed against tumor necrosis factor-alpha mRNA. J. Biol. Chem. 271: 17445-17452, 1996.

4. PubMed 18177500; Hartley JW, et al. Expression of infectious murine leukemia viruses by RAW264.7 cells, a potential complication for studies with a widely used mouse macrophage cell line. Retrovirology. 4: 5:1, 2008.

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