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D.S. Sakaguchi, J.B. Recknor, M.J. Young, S.K. Mallapragada; Growth and Differentiation of Retinal and Brain Progenitor Cells on Micropatterned Polymer Substrates . Invest. Ophthalmol. Vis. Sci. 2006;47(13):5765.
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© ARVO (1962-2015); The Authors (2016-present)
To examine the influence of physical and chemical guidance cues on growth, alignment, and differentiation of murine retinal and brain progenitor cells (RPCs and BPCs).
Micropatterned polystyrene substrates were fabricated and poly–L–lysine and laminin (LN) used to chemically modify the patterned surface and contribute substrate–associated factors for cell adhesion and directed outgrowth. The pattern dimensions used for these experiments were 16 µm groove width, 13 µm mesa width (or groove spacing) and 4 µm groove depth. Green fluorescent protein expressing RPCs and BPCs were cultured under differentiation conditions on these micropatterned substrates. Differentiation of the RPCs and BPCs was assessed morphologically and immunocytochemically after 7 days in vitro using cell–type specific antibodies.
The RPCs and BPCs aligned along the grooves of the substrate. The RPCs and BPCs appeared highly elongated and often extended their processes in the direction of the grooves and their boundaries. These results show that a synergistic combination of guidance cues facilitated oriented growth of the RPCs and BPCs. After 7 days in vitro, many of the RPCs and BPCs have expressed neuronal or glial markers, and the majority of these cells were aligned in the direction of the grooves.
Providing physical and chemical cues produces a dynamic environment for specific cellular activities such as directed growth and differentiation of neural progenitor cells. These results have important application in developing novel experimental strategies for neural regeneration.
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