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Age:
7 to 10 weeks
Comments:
The cells display prominent cytoskeletal staining for actin, and have abundant parallel fibrils in the cytoplasm. They are reported to contract in the presence of 1 X 10（-6） M angiotensin II. The cells form colonies in soft agar, and are positive for SV40 large T antigen.
Propagation:
ATCC complete growth medium: The base medium for this cell line is 3:1 mixture of ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002 and Ham's F12 medium with 14 mM HEPES.To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 5%. Temperature: 37.0°C
Subculturing:
Protocol:
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% （w/v） Trypsin- 0.53 mM 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.
Subc*tion Ratio: A subc*tion ratio of 1:4 to 1:10 is recommended Medium Renewal: Every 2 to 3 days
22754: MacKay K, et al. Glomerular epithelial, mesangial, and endothelial cell lines from transgenic mice. Kidney Int. 33: 677-684, 1988. PubMed: 2835539 51029: Robey RB, et al. Regulation of mesangial cell hexokinase activity by PKC and the classic MAPK pathway. Am. J. Physiol. 277: F742-F749, 1999. PubMed: 10564237 51030: Robey RB, et al. Thrombin is a novel regulator of hexokinase activity in mesangial cells. Kidney Int. 57: 2308-2318, 2000. PubMed: 10844601 61211: Robey RB, et al. Regulation of mesangial cell hexokinase activity and expression by heparin-binding epidermal growth factor-like growth factor: epidermal growth factors and phorbol esters increase glucose metabolism via a common mechanism involving classic mitogen-activated protein kinase pathway activation and induction of hexokinase II expression. J. Biol. Chem. 277: 14370-14378, 2002. PubMed: 11782486 61224: Maile S, et al. The morphology of mesangial cells cultured at high density and in collagen gels. Histol. Histopathol. 15: 403-414, 2000. PubMed: 10809358 61225: Coy PE, et al. LPA is a novel lipid regulator of mesangial cell hexokinase activity and HKII isoform expression. Am. J. Physiol. Renal Physiol. 283: F271-F279, 2002. PubMed: 12110510 90657: Taneja N, et al. Proinflammatory interleukin-1 cytokines increase mesangial cell hexokinase activity and hexokinase II isoform abundance. Am. J. Physiol. Cell Physiol. 287: C548-C557, 2004. PubMed: 15070811
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