Table 1.

Comparison of Original and Improved 2D-Multistep Methods

Direct comparison of media and additives and their final concentrations for the original Smith et al. (2013) (A, green) and the improved 2D-multistep (B, blue) methods. Values alone indicate a change of media, + followed by a number indicates this final concentration has been added to the existing media, and + alone indicates addition at the same concentration as added previously. The bottom row provides a summary of the changes made from the original protocol. 2PAA, 2-phospho-L-ascorbic acid; B-27 supp, B-27 supplement; bFGF, basic fibroblast growth factor; BMP4, bone morphogenetic protein 4; BSA, bovine serum albumin; EPO, erythropoietin; FICZ, 6-formylindolo(3,2-b)carbazole; FLT3L, FMS-related tyrosine kinase 3 ligand; HAMS-F12, Ham's nutrient medium F-12; [high], high concentration; IL6, interleukin-6; IMDM, Iscove's modified Dulbecco's medium; KOSR, KnockOut Serum Replacement; MTG, monothioglycerol; N-2 supp, N-2 supplement; RPMI, Roswell Park Memorial Institute medium; SCF, stem cell factor; StemPro, StemPro-34 SFM Complete Medium; TPO, thrombopoietin; VEGF, vascular endothelial growth factor A165; Wnt3a, Wingless-type MMTV integration site 3A.