User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/02/15

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Putting cells to S0
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Summary
To sensitize airway smooth muscle cells (ASMC's) to stimulation cells are put to DMEM medium without serum (S0).

Materials

 * Dulbecco's Modified Eagle Medium (DMEM) (500 mL)
 * Heat inactivated Fetal Bovine Serum (FBS) (50 mL)
 * Penicilin/Streptomycin (PenStrep) (5000 units per mL each)
 * Fungizone
 * ASMC's grown in 24-wells plate.
 * Warm PBS

Preparation of Medium
 DMEM S0   DMEM S+ 
 * 500 mL DMEM
 * 3 mL Fungizone
 * 10 mL PenStrep
 * Remove 50 mL of DMEM out of 500 mL
 * Add 50 mL FBS
 * 3 mL Fungizone
 * 10 mL PenStrep

Putting cells on S0

 * Remove DMEM S+
 * Wash twice with warm PBS, be sure to remove all medium
 * Add 500 μL DMEM S0
 * Incubate ON @ 37 °C

Run 1: Ht31/Ht31P D9/D12

 * Splitting cells 10Feb2010
 * Counting cells 12Feb2010


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