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female patient, pregnant myometrium (39 weeks gestation)

The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, (ATCC^® No. 30-2002). To make the complete growth medium, add the following components to the base medium:

• 0.1 mg/mL G-418
• additional 2 mM Glutamine
• heat-inactivated fetal bovine serum (FBS) to a final concentration of 10%

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 0.05% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which containstrypsin inhibitor.
3. Add 5 mL of 0.05% 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 1.0 X 1⁴ to 2.0 X 10⁴ viable cells/cm² is recommended.
7. Incubate cultures at 37°C.

Monga M, et al. Oxytocin-stimulated responses in a pregnant human immortalized myometrial cell line. Biol. Reprod. 55(2): 427-32, 1996. PubMed: 8828850

Yang M, et al. Multiple Trp isoforms implicated in capacitative calcium entry are expressed in human pregnant myometrium and myometrial cells. Biol Reprod 67(3):988-94, 2002. PubMed: 12193412

Zhong M., et al. Extracellular signal-regulated kinase 1/2 activation by myometrial oxytocin receptor involves Galpha(q)Gbetagamma and epidermal growth factor receptor tyrosine kinase activation. Endocrinology 144(7): 2947-56, 2003. PubMed: 12810550

Zhong M., et al. Multiple signals regulate phospholipase CBeta3 in human myometrial cells. Biol Reprod. 78(6): 1007-17, 2008. PubMed: 18322273

Murtazina DA, TRPC1, STIM1, and ORAI influence signal-regulated intracellular and endoplasmic reticulum calcium dynamics in human myometrial cells. Biol. Reprod. 85(2): 315-26, 2011. PubMed: 21565997