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Insulin-like growth factor 2 (IGF2; documented in PubMed 20404007)

• 30 nM Paclitaxel
• fetal bovine serum to a final concentration of 10%

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 1.0 to 2.0 mL Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (ATCC^® No. 30-2020) or 0.25% (w/v) Trypsin - 0.53 mM EDTA (ATCC^® No. 30-2101) solution to remove all traces of serum which contains trypsin inhibitor.
3. Add 1.0 to 2.0 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.
4. Add 6.0 to 8.0 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 10mL fresh growth medium. 
7. Add appropriate aliquots of the cell suspension to new culture vessels.
8. Incubate cultures at 37°C.

Buick RN, et al. Comparative properties of five human ovarian adenocarcinoma cell lines. Cancer Res. 45: 3668-3676, 1985. PubMed: 4016745

Huang GS, et al. Insulin-like growth factor 2 expression modulates Taxol resistance and is a candidate biomarker for reduced disease-free survival in ovarian cancer. Clin. Cancer Res. 16(11): 2999–3010, 2010. PubMed: 20404007

King ER, et al. The insulin-like growth factor 1 pathway is a potential therapeutic target for low-grade serous ovarian carcinoma. Gynecol. Oncol. 123(1): 13-18, 2011. PubMed: 21726895