DEPARTAMENTO DE CIÊNCIAS DA VIDA ... - Estudo Geral

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coated with COL-1 (similar to what was described in Engler et al., 2006), or on TCPs as
control. Cells differentiated on TCPs expressed less Beta-III (B-III) tubulin and Nestin (a
neuronal marker and an early neural marker, respectively) than the ones differentiated
on the PA hydrogels. On the other hand, there were no major differences between
cells differentiated on the ≈1kPa and ≈7kPa hydrogels (Figure 20). The expression of
the glial markers tested - O4 and GFAP (markers for oligodendrocyte and astrocyte
commitment, respectively) - was higher in the 10% PA hydrogels for GFAP, while for O4
it was higher in cells plated on TCPs and 10% PA hydrogels when compared with cells
on the softer (3% PA) hydrogels (Figure 21). In general, these results are consistent
with results obtained by others using MSCs cultured in 3D hydrogels (Her et al., 2013),
where glial fates were also favored by stiffer substrates (~10 kPa) and neuronal and
neural precursor markers were favored by softer substrates (~1 kPa).
Figure 20 – hMSCs cultured on TCPs (Plastic) and PA hydrogels coated with COL-1 for 7 days after being
treated with mitomycin C to inhibit proliferation. Cells were stained with anti-B-III tubulin (red), anti-Nestin
antibodies (green) and DAPI (blue). This experiment was performed once (n=1). Bar corresponds to 400µm.