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

Summary

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

Materials & Methods

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

Method

Preparation of Medium

DMEM S⁰

• 500 mL DMEM
• 3 mL Fungizone
• 10 mL PenStrep

DMEM S⁺

• 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

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