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. Author manuscript; available in PMC: 2018 Aug 1.
Published in final edited form as: Exp Eye Res. 2017 May 30;161:17–29. doi: 10.1016/j.exer.2017.05.011

Table 3.

Growth medium for retinal cell lines*

Name Source Catalogue number Concentration (or proportional volume) per liter in final medium# Comments
DMEM with HEPES Sigma-Aldrich, Saint Louis, MO D6171 460 ml High glucose formulation
Ham’s F-12 with HEPES Sigma-Aldrich N8641 450 ml
TAPSO Sigma-Aldrich T9269 1 g Supplemental pH buffering component
HCl Sigma-Aldrich H9892 2 ml Supplied as sterile 1 N solution
Bovine calf serum HyClone, Logan, UT SH30073.03 2 ml Filter-sterilize thawed aliquots and store refrigerated not more than 2 wk
Vitamin E acetate Sigma-Aldrich T3001 1 mg From ethanol stock
Cholesterol Sigma-Aldrich C8667 2 mg From ethanol stock
Bovine retina extract Animal Technologies, Tyler, TX (retinas) (N/A) 2.5 ml Refer to Pfeffer (1990) for details of preparation; final amount of added protein approximately
6 mg/L; thawed aliquots stored refrigerated not more than 1 wk
Linoleic acid
(complexed with bovine serum albumin (BSA))		 Cayman Chemical, Ann Arbor, MI;
(Sigma-Aldrich)		 90150
(A8806)		 1 mg
(120 mg)		 Linoleic acid stored under Ar at −20°C;
(Complex stored under Ar at −80°C)
Transferrin Sigma-Aldrich T8158 7.5 mg Solid and stock aliquots both stored at −20°C
Long® R3 IGF-1 Sigma-Aldrich I1271 10 µg
Oxalacetic acid Sigma-Aldrich O7753 66 mg
Sodium pyruvate Sigma-Aldrich S8636 5 ml Supplemental; supplied as sterile 100 mM solution; store refrigerated
Sodium selenite Sigma-Aldrich S9133 0.173 µg
MnCl2•4H2O Sigma-Aldrich M3634 0.9 µg
(NH4)6Mo7O24•4H2O Sigma-Aldrich M1019 1.24 µg
Glucose Sigma-Aldrich G7021 375 mg Supplemental; stock formulated together with the next four components (below)
Fructose Sigma-Aldrich F3510 375 mg
D-(+)-Gluconic acid lactone Sigma-Aldrich G4750 10 mg
D-Glucuronic acid, sodium salt monohydrate Sigma-Aldrich G8645 1 mg
D-(+)-Glucosamine HCl Sigma-Aldrich G1514 5 mg
D-(+)-Galactose Sigma-Aldrich G0750 180 mg Stock formulated with the next two components (below)
L-(−)-Fucose Sigma-Aldrich F2252 82 mg
β-Lactose Sigma-Aldrich L3750 750 mg
Non-essential amino acids Sigma-Aldrich M7145 5 ml Supplement supplied as sterile solution; refrigerate
Taurine Sigma-Aldrich T8691 1.25 mg
Heparin, sodium salt Sigma-Aldrich H3149 1 mg Not less than 175 U per mg solid
Alanyl-glutamine Sigma-Aldrich A8185 868 mg
Uridine Sigma-Aldrich U3003 0.3 mg
Thymidine Santa Cruz Biotechnology,
Dallas, TX		 296542 1 mg Supplemental
L-Carnitine tartrate LKT Laboratories, St. Paul, MN C0264 2.35 mg
Thiamine hydrochloride Santa Cruz Biotechnology 205859 4 mg Supplemental
Ethanolamine Sigma-Aldrich E0135 0.5 mg Viscous liquid stored at rt
Glutathione SH Sigma-Aldrich G6529 2 mg
Hydrocortisone Sigma-Aldrich H0396 6 µg Complexed with cyclodextrin; amount given with respect to hydrocortisone
Progesterone Sigma-Aldrich P7556 35 µg Complexed with cyclodextrin; amount given with respect to progesterone
Retinyl acetate Sigma-Aldrich R0635 0.2 mg Complexed with cyclodextrin; amount given with respect to retinyl acetate; protect from light
Triiodothyronine Sigma-Aldrich T6397 3.5 µg
Ascorbic acid phosphate Wako Chemicals, Richmond, VA 013-19641 36 mg
Putrescine Sigma-Aldrich P5780 0.8 mg Supplemental
Thioglycerol Sigma-Aldrich M6145 5.4 mg Supplied as liquid; store refrigerated and tightly sealed
*

Use with pCO2 = 6%

#

All components expressed by weight are added from concentrated stock solutions