| 產(chǎn)品名稱 |
RMC |
| 商品貨號(hào) |
B165614 |
| Organism |
Rattus norvegicus, rat |
| Tissue |
kidney |
| Cell Type |
Mesangial Cell |
| Product Format |
frozen |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain SV40 viral sequences]
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 |
3 months |
| Gender |
male |
| Strain |
Sprague-Dawley |
| Applications |
Mesangial cells are crucial for kidney function and are used extensively in kidney research.
|
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
Rat mesangial cell at passage 8 were immortalized with pSV3-Neo and maintained in the presence of G418. |
| Clinical Data |
male
3 months |
| Genes Expressed |
desmin; vimentin |
| Cellular Products |
desmin; vimentin |
| Comments |
Rat mesangial cell at passage 8 were immortalized with pSV3-Neo and maintained in the presence of G418.
The cells express normal genes of the wild type mesangial cells; they are positive for desmin and vimentin and negative for cytokeratin 8.
|
| Complete Growth Medium |
Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose and supplemented with 0.4 mg/ml G418, 85%; fetal bovine serum, 15%
|
| 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.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53% (w/v) EDTA solution to remove all traces of serum that contains trypsin inhibitor.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C
Subculture Ratio: 1:4 to 1:8
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 |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Name of Depositor |
M Ailenberg, M Silverman |
| Deposited As |
rat |
| Passage History |
Rat mesangial cell at passage 8 were immortalized with pSV3-Neo and maintained in the presence of G418. |
