国产精品国产精品一区精品国产自在现偷99精品国产在热2019国产拍偷精品网国产精品视频全国免费观看,国产精品v欧美精品v日韩精品青青精品视频国产久久国产精品久久精品国产亚洲精品国产精品国产欧美精品一区二区三区,国产精品第一页国产亚洲精品国产福利国产精品自拍国产精品视频在线观看亚洲国产精品一区二区久久国产精品国产三级国产专不,国产精品视频大陆精大陆国产国语精品2019精品国产品对白在线285年香蕉精品国产高清自在自线隔壁老王国产在线精品在线观看精品国产福利片,国产三级精品三级在专区精品国产自在现偷国产精品一区二区三区国产日韩精品欧美一区喷水亚洲精品国产精品国自产国产在线精品一区二区不卡

1. Remove and discard culture medium.
2. Briefly rinse the cell layer, twice, with 5 mL Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
3. Add 5 mL of 0.25% 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 5 mL of complete growth medium and aspirate cells by gently pipetting.
5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new vessels. An inoculum of 7.0 X 1⁴ to 1.0 X 10⁵ viable cells/cm² is recommended.
7. Incubate cultures at 37°C.

Muszynski M, et al. Glucocorticoids stimulate the production of preprocalcitonin-derived secretory peptides by a rat medullary thyroid carcinoma cell line. J. Biol. Chem. 258(19): 11678-83, 1983. PubMed: 6311819

Birnbaum RS, et al. Identification of procalcitonin in a rat medullary thyroid carcinoma cell line. J. Biol. Chem. 259(5): 2870-4, 1984. PubMed: 6199352

Russo AF, et al. Neuronal properties of a thyroid C-cell line: partial repression by dexamethasone and retinoic acid. Mol. Endocrinol. 6(2): 207-18, 1992. PubMed: 1569964

Birnbaum RS, et al. Biosynthesis of calcitonin by a rat medullary thyroid carcinoma cell line. J. Biol. Chem. 261(2): 699-703, 1986. PubMed: 3941098

Durham PL, et al. Repression of the calcitonin gene-related peptide promoter by 5-HT1 receptor activation. J. Neurosci. 17(24): 9545-53, 1997. PubMed: 9391009

Wood JL, et al. Autoregulation of cell-specific MAP kinase control of the tryptophan hydroxylase promoter. J. Biol. Chem. 276(24): 21262-71, 2001. PubMed: 11283010

Viney TJ, et al. Regulation of the cell-specific calcitonin/calcitonin gene-related peptide enhancer by USF and the Foxa2 forkhead protein. J. Biol. Chem. 279(48): 49948-55, 2004. PubMed: 15385550

Park KY, et al. Control of the calcitonin gene-related peptide enhancer by upstream stimulatory factor in trigeminal ganglion neurons. J. Biol. Chem. 283(9):5 441-51, 2008. PubMed: 18167349