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Stem Cells International
Volume 2016 (2016), Article ID 2452985, 8 pages
Research Article

Modeling Neurological Disease by Rapid Conversion of Human Urine Cells into Functional Neurons

1Department of Neurology, Institute of Neurology, Huashan Hospital, Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Shanghai Medical College, Fudan University, Shanghai 200040, China
2Department of Neurology, Research Center of Neurology in Second Affiliated Hospital, and the Collaborative Innovation Center for Brain Science, Zhejiang University School of Medicine, Hangzhou 310009, China
3Department of Anatomy, Histology & Embryology, Shanghai Medical College, Fudan University, Shanghai 200030, China
4Institutes of Brain Science, Institute of Neurobiology and State Key Laboratory of Medical Neurobiology, Fudan University, Shanghai 200030, China

Received 11 April 2015; Revised 18 July 2015; Accepted 22 July 2015

Academic Editor: Gary E. Lyons

Copyright © 2016 Shu-Zhen Zhang et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

In order to convert urine cells into functional neurons, five retroviruses carrying Ascl1, Brn2, NeuroD, c-Myc, and Myt1l were used. Twenty-four hours after the recovery, the five retroviruses were added to primary urinary cells for one day. The medium was changed to EPi/N2 medium supplied with 1 µg/ml DOX the second day and last for 2 days. Then cells were replaced onto astrocyte coated cover-slips in N2 medium with supplementary factors. The culture medium was changed every day until used.

The morphology changes of the urine cells during this procedure were described here. For the first 4 days, the cells showed epithelial-like morphology and sustained proliferation. From the Day 4, cells began to change their shape. The expression of the transcriptional factors was analyzed on Day 4. We found that almost all the cells expressed GFP, but we could not know which cells expressed the 5 factors at the same time. On the Day 5, about 30% of the cells elongated and became long spindle cells. Some grew dendrite-like structures. Unfortunately, only a small percentage of these cells could be converted into neurons. Most of these cells began to die at almost the same time. Cells which were successfully converted into neurons grew long processes and exhibited neuron-like morphology. And these cells could be labeled by neuron lineage marker Tuj1.

  1. Supplementary Material