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Age:
embryo
Comments:
SPARC is an extracellular Ca+2 binding glycoprotein, which is differentially associated with morphogeneisis, remodeling, cellular migration and proliferation [PubMed; 2745554].Exogenous SPARC, added to cells in culture is associated with profound changes in cell shape, caused rapid, partial detachment of a confluent monolayer, and inhibited spreading of newly plated cells [PubMed; 2745554].
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Atmosphere: air, 95%; carbon dioxide (CO2), 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.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended Medium Renewal: Every 2 to 3 days
Preservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002 recommended serum:ATCC 30-2020
References:
57812: Sage H, et al. SPARC, a secreted protein associated with cellular proliferation, inhibits cell spreading in vitro and exhibits Ca+2-dependent binding to the extracellular matrix. J. Cell Biol. 109: 341-356, 1989. PubMed: 2745554 61189: Basu A, et al. The intracisternal A-particle proximal enhancer-binding protein activates transcription and is identical to the RNA- and DNA-binding protein p54nrb/NonO. Mol. Cell. Biol. 17: 677-686, 1997. PubMed: 9001221 61190: Satyamoorthy K, et al. Identification of an intronic enhancer that nullifies upstream repression of SPARC gene expression. Nucleic Acids Res. 25: 3169-3174, 1997. PubMed: 9224620 89731: Lehman JM, et al. Neoplastic differentiation: characteristics of cell lines derived from a murine teratocarcinoma. J. Cell. Physiol. : 13-27, 1974. PubMed: 4858519