DB1X

The clonal cell line A10 was derived by B. Kimes and B. Brandt from the thoracic aorta of DB1X embryonic rat and possesses many of the properties characteristic of smooth muscle cells.
The cells produce spontaneous action potentials at the stationary phase of the growth cycle and exhibit an increase in activity of the enzymes myokinase and creatine phosphokinase.

Protocol:

1. Remove and discard culture medium.
2. 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.
3. 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.
4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.

Subcultivation ratio: A subcultivation ratio of 1:3 to 1:6 is recommended

Medium renewal: Every 3 to 4 days

29 hours

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