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Contents

Background

These cells were made from ~0.5g chunks of kidney taken from a male with ADPKD.
Refer to (Making Primary Cells from Mouse Tissue protocol) There were 9 different tissue samples in total. Which part of the kidney the tissue was taken from is unknown as the kidney was so riddled with cyts and fibrotic tissue that the original kidney anatomy was unrecognisable.
Six samples successfully grew in culture. Samples are identified as "sample 1" etc.

Grow Cells on Collagen Coated Surfaces

Refer to (Collagen Coating of Cell Culture Vessels) for protocol.

Cell Culture Media used for human ADPKD kidney cells (with additives)

Reagent 50mL 500mL Final Concentration
DMEM/F-12 (1:1) 46.85mL
insulin-transferrin-selenium (100x) 375uL3.75mL0.75x
dexamethasone (20ug/mL) 250uL2.5mL100ng/mL
triiodotyronine (20ug/mL) 15uL150uL6ng/uL
murine EGF (0.1mg/mL) 5uL50uL10ng/mL
pen/strep 0.5mL5mL
FCS 2.5mL25mL5%



Insulin-Transferrin-Selenium (100x)

Insulin-transferrin-selenium, 10mL, Invitrogen #41400-045
Store at 4 degrees C
Ready to use

Dexamethasone (20ug/mL)

Dexamethasone, 1mg, Sigma #D8893
Lyophilised reagent stored at 4 degrees C


Triiodotyronine (20ug/mL)

Triiodotyronine, 1mg, Sigma #T5516


Murine Epidermal Growth Factor

mEGF, 0.1mg, Sigma #E4127