DEPARTAMENTO DE CIÊNCIAS DA VIDA ... - Estudo Geral

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I.4 - Objectives
It is of interest to understand what would be the effect of isolating MSCs directly
on substrates with stiffness similar to that of neural tissues in terms of their potential
to express neural markers.
We propose to isolate and culture human umbilical cord matrix MSCs directly on
softer substrates, namely hydrogels compliant with neural tissue (1 to 10KPa) and, as a
control, part of the umbilical cord matrix of every sample will be used to isolate MSCs
using normal tissue-culture polystyrene plates (the typical isolation and culture
protocol) and then transferred onto similar hydrogels after several passages on
polystyrene (P1-P5). To study if prolonged culture of MSCs on stiff polystyrene (P2-P5)
will restrain their capacity to express neural markers later on or not when compared to
isolated cells in the hydrogels. We will then assess the differences in expression levels
of specific MSCs markers, but also of makers of neural stem/progenitor cells (nestin)
and specific neural lineage markers, such as neuronal (beta-III-tubulin),
oligodendroglial (O4) and astroglial (GFAP) markers.