Article

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

ABSTRACT

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.

Download full-text

Full-text

Available from: Dajiang Qin, Apr 23, 2014
  • Source
    • "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. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Fibrodysplasia ossificans progressiva (FOP) is a rare disease characterized by progressive ossification of soft tissues, for which there is no effective treatment. Mutations in the bone morphogenetic protein (BMP) type I receptor activin receptor-like kinase 2 (ACVR1/ALK2) are the main cause of FOP. We generated human induced pluripotent stem cells (hiPSCs) from FOP patients with the ALK2 R206H mutation. The mutant ALK2 gene changed differentiation efficiencies of hiPSCs into FOP bone-forming progenitors: endothelial cells (ECs) and pericytes. ECs from FOP hiPSCs showed reduced expression of vascular endothelial growth factor receptor 2 and could transform into mesenchymal cells through endothelial-mesenchymal transition. Increased mineralization of pericytes from FOP hiPSCs could be partly inhibited by the ALK2 kinase inhibitor LDN-212854. Thus, differentiated FOP hiPSCs recapitulate some aspects of the disease phenotype in vitro, and they could be instrumental in further elucidating underlying mechanisms of FOP and development of therapeutic drug candidates.
    Full-text · Article · Nov 2015 · Stem Cell Reports
  • Source
    • "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). "
    [Show abstract] [Hide abstract]
    ABSTRACT: In mice, leukemia inhibitory factor (LIF)-dependent primitive neural stem cells (NSCs) have a higher neurogenic potential than bFGF-dependent definitive NSCs. Therefore, expandable primitive NSCs are required for research and for the development of therapeutic strategies for neurological diseases. There is a dearth of suitable techniques for the generation of human long-term expandable primitive NSCs. Here, we have described a method for the conversion of human fibroblasts to LIF-dependent primitive NSCs using a strategy based on techniques for the generation of induced pluripotent stem cells (iPSCs). These LIF-dependent induced NSCs (LD-iNSCs) can be expanded for >100 passages. Long-term cultured LD-iNSCs demonstrated multipotent neural differentiation potential and could generate motor neurons and dopaminergic neurons, as well as astrocytes and oligodendrocytes, indicating a high level of plasticity. Furthermore, LD-iNSCs easily reverted to human iPSCs, indicating that LD-iNSCs are in an intermediate iPSC state. This method may facilitate the generation of patient-specific human neurons for studies and treatment of neurodegenerative diseases.
    Preview · Article · Oct 2015 · Biology Open
  • Source
    • "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. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Granule neurons in the hippocampal dentate gyrus (DG) receive their primary inputs from the cortex and are known to be continuously generated throughout adult life. Ongoing integration of newborn neurons into the existing hippocampal neural circuitry provides enhanced neuroplasticity, which plays a crucial role in learning and memory; deficits in this process have been associated with cognitive decline under neuropathological conditions. In this Primer, we summarize the developmental principles that regulate the process of DG neurogenesis and discuss recent advances in harnessing these developmental cues to generate DG granule neurons from human pluripotent stem cells.
    Full-text · Article · Jun 2014 · Development
Show more