When grown in osteogenic medium, the cells expressed alkaline phosphatase (ALP) and osteocalcin mRNA.
After the removal of the cell medium, the cells were washed by 70% cold methanol for 15 minutes on shaking.
After removing the medium, the cells were washed three times with PBS in order to detach the free csMSN.
When they put human macrophages in the lower compartment filled with cell culture medium, the cells rose toward the membrane.
When all factors were added to the culture medium, the cells exhibited features that closely resembled human adult hepatocytes (Chivu et al., 2009).
Most interestingly, when isolated perivascular MSCs were exposed to glial induction medium, the cells differentiated into oligodendrocytes or astrocytes, pericyte-specific antigens were downregulated and cells expressed glial fibrillary acidic protein (GFAP).
When differentiated with 2.5 μM 5Brdu in a low serum (2%) medium, the cells stops proliferating and forms extensive network of neurites (red arrows in Fig. 2b) together with connected clumps of cells (green arrow in Fig. 2b).
Using the new medium, the cells endocytosed ligands for up to 20 days, and survived for at least an additional 10 days, albeit without the high endocytic activity typical of intact LSECs.
After exchange of the medium, the cells were incubated for 60 hours.
Upon addition of nocodazole to the growth medium the cells arrested at the SAC and started contracting (Movie S8B).
After 5 days in drug-containing medium, the cells were grown in drug-free medium for another 3 5 days before further studies were performed.
