Octanoate in Human Albumin Preparations Is Detrimental to Mesenchymal Stromal Cell Culture

Figure 4

The effects of octanoate on MSCs. Growth of MSCs is inhibited at levels of octanoate in albumin products that are relevant to cell culture. Cells were grown for 3 days in PPRF-msc6 media with C-NPOF albumin and varying amounts of octanoate before counting. An inhibitory effect of 50% is seen at octanoate levels between 0.5 and 0.6 mM (a). Colony formation is reduced to about 40% of cells grown in PPRF-msc6 with C-NPOF albumin, 0.67 mM octanoate for 4 passages (b). MSCs grown for 4 passages in PPRF-msc6 with C-NPOF albumin ((i), (ii)) and C-NPOF albumin with octanoate ((iii), (iv)) express Na⁺/K⁺ ATPase. Panels (ii) and (iv) contain mouse anti-Na⁺/K⁺ ATPase 1 : 50 while all panels contain secondary goat anti-mouse antibody coupled to Alexa Fluor 488 (green). Nuclei are stained blue with Hoechst 33258. Bar equals 20 μm (c). Quantitation of Na⁺/K⁺ ATPase expression in MSCs. Cells were grown for four passages as in (c) before being lysed in RIPA buffer and analyzed by Western blotting. Bands were quantitated using Image-J and values normalized against β-actin. Experiments were performed in duplicate (d). ** indicates a .