User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/03/12

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


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Summary

 * HT4 cells split 10Feb2010 were stored @ -80°C by Anouk
 * Splitting hTERT
 * Refreshing media

Materials

 * DMEM S+
 * Remove 50 mL of DMEM out of 500 mL
 * Add 50 mL FBS
 * 3 mL Fungizone
 * 10 mL PenStrep
 * hTERT Cells
 * D12 P26 (9Feb2010)
 * D9 P22 (9Feb2010)

Method

 * Wash cells twice with warm PBS
 * Add 1 mL Trypsin
 * Incubate @ 37 °C
 * Collect cells in total volume of 30 mL DMEM S+
 * Count cells

Results

 * D12 (66×104 cells / mL)
 * 74
 * 67
 * 69
 * 54
 * For 24-wells 1.36 mL cells in 11.14 mL S+, for 3 petridishes 4.55 mL cells in 25.45 mL S+
 * D9 (75×104 cells / mL)
 * 74
 * 58
 * 77
 * 64
 * For 24-wells 1 mL cells in 12.5 mL S+, for 3 petridishes 4 mL cells in 26 mL S+

Related

 * Splitting cells
 * Counting cells
 * History cells


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