Efficient induction of neural precursor cells from fibroblasts using stromal cell-derived inducing activity

Title
Efficient induction of neural precursor cells from fibroblasts using stromal cell-derived inducing activity
Authors
임미선
Keywords
Induced neural precursor cell; Direct conversion; Stromal cell; Co-culture; Dopaminergic neuron
Issue Date
2016-02
Publisher
KOREAN TISSUE ENGINEERING REGENERATIVE MEDICINE SOC
Citation
TISSUE ENGINEERING AND REGENERATIVE MEDICINE, v. 13, NO 5, Page. 554-559
Abstract
The direct lineage conversion of fibroblasts into neuronal or neural precursor cells (NPCs) has become a hot issue in recent years as an attractive approach in the field of stem cell regenerative medicine. In this study, we adopted the stromal feeder co-culture method during the early conversion period to enhance conversion efficiency. Stromal cells are often used in directed differentiation of dopaminergic (DA) neurons from pluripotent stem cells. We co-cultured rat embryonic fibroblasts (REFs) on gamma-irradiated sonic hedgehog-overexpressing MS5 stromal (MS5-SHH) cells after transduction with Brn2, Ascl1, Myt1L, and BclxL-GFP (BAMX(GFP)) transcription factors to REFs. One week after co-culture, transduced cells (GFP+ cells) that proliferated on MS5-SHH cells were separated from MS5-SHH cells through a 40 A mu m cell strainer. Subsequently, the converted cells (GFP+ cells) were expanded on fibronectin-coated culture plates in NPC expansion medium. The induced NPCs (iNPCs) expressed NPC potential (NESTIN+/SOX2+) earlier than seen with non-co-culture methods and were efficiently differentiated into DA neurons by overexpression of Nurr1 and Foxa2 genes, which are specific transcription factors for midbrain DA neuron development. These observations indicated that direct conversion to NPCs using an MS5 stromal cells co-culture method is a suitable technique for efficient generation of iNPC/DA neurons from fibroblasts.
URI
https://link.springer.com/article/10.1007%2Fs13770-016-0012-3http://hdl.handle.net/20.500.11754/33859
ISSN
1738-2696
DOI
http://dx.doi.org/10.1007/s13770-016-0012-3
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RESEARCH INSTITUTE[S](부설연구소) > ETC
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