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Microsoft Word - MediaRecipesForMCF10ACells

The document describes three different media recipes for culturing MCF-10A cells, including growth medium, resuspension medium, and assay medium. It lists the components, amounts, and concentrations needed for each type of medium, noting that some components like EGF and hydrocortisone vary between the recipes. Instructions are provided for reconstituting and storing the additives.

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Farhana Anuar
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166 views

Microsoft Word - MediaRecipesForMCF10ACells

The document describes three different media recipes for culturing MCF-10A cells, including growth medium, resuspension medium, and assay medium. It lists the components, amounts, and concentrations needed for each type of medium, noting that some components like EGF and hydrocortisone vary between the recipes. Instructions are provided for reconstituting and storing the additives.

Uploaded by

Farhana Anuar
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Table 1: Media recipes for MCF-10A Cells

Component Growth Medium1 Resuspension Medium1 Assay Medium1


(Without EGF)
DMEM/F12 500.0 ml 400.0 ml 500.0 ml
(Invitrogen #11330-032)
Horse Serum 25.0 ml 100.0 ml 10.00 ml
(Invitrogen#16050-122) (5% final) (20% final) (2% final)
EGF 100 μl -- --
(100μg/ml stock)2 (20ng/ml final)
Hydrocortisone 250 μl -- 250 μl
(1mg/ml)3 (0.5 mg/ml final) (0.5 μg/ml final)
Cholera Toxin 50 μl -- 50μl
(1mg/ml stock)4 (100 ng/ml final) (100 ng/ml final)
Insulin 500 μl -- 500μl
(10mg/ml stock)5 (10μg/ml final) (10μg/ml final)
Pen/Strep 5.0 ml 5.0 ml 5.0 ml
(100x solution, Invitrogen #15070-063)

Notes:
1) For each medium type, premix all of the appropriate additives, sterile filter through a 0.2 μm filer, and add to DMEM/F12 medium
bottle.
2) EGF: (Peprotech, 1 mg): Resuspend at 100 μg/ml in sterile dH2O. Store aliquots at –20oC.
3) Hydrocortisone: (Sigma #H-0888, 1 g bottles) Resuspend at 1 mg/ml in 200 proof ethanol and store aliquots at -20 oC.
4) Cholera Toxin: (Sigma #C-8052, 2 mg vials) Resuspend at 1 mg/ml in sterile dH2O and allow to reconstitute for about 10 minutes.
Store aliquots at 4oC.
5) Insulin: (Sigma #I-1882, 100 mg vials) Resuspend at 10 mg/ml in sterile dH2O containing 1% glacial acetic acid. Shake solution
and allow 10-15 min to reconstitute. Store aliquots at –20C.

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