Tissue Eng Regen Med.
2016 Oct;13(5):554-559. 10.1007/s13770-016-0012-3.
Efficient Induction of Neural Precursor Cells from Fibroblasts Using Stromal Cell-Derived Inducing Activity
- Affiliations
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- 1Hanyang Biomedical Research Institute, College of Medicine, Hanyang University, Seoul, Korea. chshpark@hanyang.ac.kr
- 2R&D Center Jeil Pharmaceutical Co. Ltd., Yongin, Korea.
- 3Department of Biomedical Science, Graduate School, Hanyang University, Seoul, Korea.
- 4Graduate School of Biomedical Science and Engineering, Hanyang University, Seoul, Korea.
- 5Department of Microbiology, College of Medicine, Hanyang University, Seoul, Korea.
- KMID: 2360804
- DOI: http://doi.org/10.1007/s13770-016-0012-3
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 γ-irradiated sonic hedgehog-overexpressing MS5 stromal (MS5-SHH) cells after transduction with Brn2, Ascl1, Myt1L, and BclxL-GFP (BAMXGFP) 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 µm 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.
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