MesenCult-ACF Plus Culture Kit

The MesenCult-ACF Plus Culture Kit from STEMCELL Technologies is an animal component-free system used for the culture of mesenchymal stromal cells (MSCs). As described in an independent protocol, the kit's medium is completed by combining MesenCult-ACF Plus Medium with MesenCult-ACF Plus 500X Supplement and 2 mM L-glutamine, and culture vessels are pre-coated with an Animal Component-Free Cell Attachment Substrate diluted in D-PBS.

Using round-bottom 96-well plates, MSCs seeded in the completed medium form visible spheroids within 1–3 days under standard incubation at 37 °C, 5% CO2, and 80% humidity. These spheroids are suitable for subsequent endothelial differentiation in medium supplemented with 10 ng/ml VEGF, EGF, and bFGF plus 2% (v/v) human serum for 5–11 days. Differentiated spheroids have been shown by immunofluorescence to express endothelial markers including von Willebrand factor (vWF) and CD31, and spheroids can be disassociated with accutase for downstream flow cytometry analysis.