Int J Stem Cells.  2019 Jul;12(2):340-346. 10.15283/ijsc18123.

Hybrid Nanofiber Scaffold-Based Direct Conversion of Neural Precursor Cells/Dopamine Neurons

• Lim MS
• Ko SH
• Kim MS
• Lee B
• Jung HS
• Kim K
• Park CH

• KMID: 2465903
• DOI: http://doi.org/10.15283/ijsc18123

Abstract

The concept of cellular reprogramming was developed to generate induced neural precursor cells (iNPCs)/dopaminergic (iDA) neurons using diverse approaches. Here, we investigated the effects of various nanoscale scaffolds (fiber, dot, and line) on iNPC/iDA differentiation by direct reprogramming. The generation and maturation of iDA neurons (microtubule-associated protein 2-positive and tyrosine hydroxylase-positive) and iNPCs (NESTIN-positive and SOX2-positive) increased on fiber and dot scaffolds as compared to that of the flat (control) scaffold. This study demonstrates that nanotopographical environments are suitable for direct differentiation methods and may improve the differentiation efficiency.

Keyword

Hybrid nanofiber; Direct conversion; Neural precursor; iNPC

MeSH Terms

Cellular Reprogramming
Nanofibers*
Neurons*
Tyrosine
Tyrosine

Figure

• 

  Fig. 1 Fabrication process of the hybrid nanofiber scaffold and planar SEM images of the fabricated nanopatterns by UV-assisted capillary force lithography (CFL). Scale bars represent 5 and 1 μm in the panel and inset images: (A) Electrospinning process to secure aligned nanofibers and transfer onto the film, (B) Image of the nanofiber based sieve for cell sheet (transfer number: 6, density: 173 fiber/mm) with PDMS frame (20 mm diameter hole, frame size: 30 mm×30 mm, height: 5 mm) and (C) 400 nm diameter and 400 nm pitch dots (left) and 400 nm width and 400 nm pitch lines (right).

• 

  Fig. 2 Generation of iNPC/iDA neurons from fibroblasts on nanoscale pattern substrates. (A) Schematic diagram depicting the timeline of conversion process. (B) Phase-contrast images of transduced cells on each nanoscale pattern and calculation graph. Scale bar=20 μm.

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  Fig. 3 Characterization of iNPCs among diverse nanoscale pattern surfaces. (A) Phase-contrast images of transduced cells and immunostained image of NPC markers, nestin and SOX2, on each nanoscale pattern. White dot box indicates high magnification images. (B) Calculation graph. Error bars denote the standard error of the mean (SEM, n=3, *p<0.05, **p<0.01). Scale bar=20 μm. D: Dot, L: Line.

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  Fig. 4 Characterization of iNPC-derived DA neurons among diverse nanoscale pattern surfaces. (A) Immunostained image of MAP2 and TH on each nanoscale pattern. (B) Calculation graph. Error bars denote the standard error of the mean (SEM, n=3, *p<0.05). Scale bar=20 μm. D: Dot, L: Line.

Reference

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