The effect of substrate topography on direct reprogramming of fibroblasts to induced neurons

被引：77
作者：
Kulangara, Karina [1 ]
Adler, Andrew F. [1 ]
Wang, Hong [2 ]
Chellappan, Malathi [1 ]
Hammett, Ellen [1 ]
Yasuda, Ryohei [2 ,3 ]
Leong, Kam W. [1 ]
机构：
[1] Dept Biomed Engn, Durham, NC 27708 USA
[2] Dept Neurobiol, Durham, NC 27708 USA
[3] Duke Univ, Howard Hughes Med Inst, Durham, NC 27708 USA
来源：
BIOMATERIALS | 2014年 / 35卷 / 20期
关键词：
Topography; Direct reprogramming; Biomaterials; Induced Neurons; DIRECT CONVERSION; DOPAMINERGIC-NEURONS; MOUSE; CELLS; ADHESION; GROWTH; GENERATION; OUTGROWTH; SURFACES;
D O I：
10.1016/j.biomaterials.2014.03.034
中图分类号：
R318 [生物医学工程];
学科分类号：
0831 ;
摘要：
Cellular reprogramming holds tremendous potential for cell therapy and regenerative medicine. Recently, fibroblasts have been directly converted into induced neurons (iNs) by overexpression of the neuronal transcription factors Ascl1, Brn2 and Myt1L. Hypothesizing that cell topography interactions could influence the fibroblast-to-neuron reprogramming process, we investigated the effects of various topographies on iNs produced by direct reprogramming. Final iN purity and conversion efficiency were increased on micrograting substrates. Neurite branching was increased on microposts and decreased on microgratings, with a simplified dendritic arbor characterized by the reduction of MAP2(+) neurites. Neurite outgrowth increased significantly on various topographies. DNA microarray analysis detected 20 differentially expressed genes in iNs reprogrammed on smooth versus microgratings, and quantitative PCR (qPCR) confirmed the upregulation of Vip and downregulation of Thy1 and Bmp5 on microgratings. Electrophysiology and calcium imaging verified the functionality of these iNs. This study demonstrates the potential of applying topographical cues to optimize cellular reprogramming. (c) 2014 Elsevier Ltd. All rights reserved.
引用
收藏
页码：5327 / 5336
页数：10
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