Generation of integration-free neural progenitor cells from cells in human urine

1] Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China. [2] Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China. [3] Department of Pathology, Dalian Medical University, Dalian, China.
Nature Methods (Impact Factor: 32.07). 12/2012; 10(1). DOI: 10.1038/nmeth.2283
Source: PubMed


Human neural stem cells hold great promise for research and therapy in neural disease. We describe the generation of integration-free and expandable human neural progenitor cells (NPCs). We combined an episomal system to deliver reprogramming factors with a chemically defined culture medium to reprogram epithelial-like cells from human urine into NPCs (hUiNPCs). These transgene-free hUiNPCs can self-renew and can differentiate into multiple functional neuronal subtypes and glial cells in vitro. Although functional in vivo analysis is still needed, we report that the cells survive and differentiate upon transplant into newborn rat brain.

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Available from: Dajiang Qin, Apr 23, 2014
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    • "In summary, cultured cells from urine were electroporated with episomal vectors containing OCT4, SOX2, KLF4, and the pCEP4-miR-302-367 cluster (containing miR-302b, c, a, d, and miR-367) (Xue et al., 2013). Transfected urine cells were maintained in serum-free mTesR1 medium supplemented with a cocktail of small molecule inhibitors to promote reprogramming: CHIR99021, PD0325901, A83-01, and thiazovivin (Wang et al., 2013). Small colonies of cells appeared that progressively adopted a human embryonic stem cell (hESC)-like morphology. "
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    • "This indirect approach would allow for the production of sufficient amounts of cells for basic research, drug screening, disease modeling, or cell therapy. Some researchers have recently reported that multipotent neural stem/progenitor cells (NSCs) can be directly induced from fibroblasts using neural progenitor-specific transcription factors or the Yamanaka factors as reprogramming agents (Han et al., 2012; Kim et al., 2011; Lujan et al., 2012; Ring et al., 2012; Thier et al., 2012; Wang et al., 2013). These induced neural stem/progenitor cells (iNSCs) are cultured in a standard neural stem cell medium supplemented with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). "
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    • "In a single culture, these may include dopaminergic neurons marked by tyrosine hydroxylase (TH), glutamatergic neurons marked by vesicular glutamate transporter, interneurons marked by γ-aminobutyric acid, and motor neurons marked by choline acetyltransferase (Ambasudhan et al., 2011; Pang et al., 2011; Qiang et al., 2011; Yoo et al., 2011). In a similar way, neurons differentiated from iNPCs thus far also seem to contain multiple neuronal subtypes (Han et al., 2012; Ring et al., 2012; Wang et al., 2013; Zou et al., 2014). It is clear from these studies that the method of NPC production and the precise patterning cues that are provided have a direct bearing on the potential of the cells to reliably differentiate into specialized cell types. "
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