Key Spec Table
|Description||ReNcell® VM Human Neural Progenitor Cell LIne|
|Overview||ReNcell® VM is an immortalized human neural progenitor cell line with the ability to readily differentiate into neurons and glial cells. ReNcell® VM was derived from the ventral mesencephalon region of human fetal brain. Immortalized by retroviral transduction with the v-myc oncogene, this cell line grows rapidly as a monolayer on laminin with a doubling time of 20-30 hours. Karyotype analyses indicate that the ReNcell® VM retains a normal diploid karyotype in culture even after prolonged passage (>45 passages). ReNcell® VM was developed by the ReNeuron Group plc, a biotech company that specializes in using human somatic stem cells for therapeutics. In experiments performed by the ReNeuron Group plc, ReNcell® VM can be differentiated in vitro to a high level of human dopaminergic neurons. Neurons differentiated from ReNcell® VM have furthermore been shown to be electophysiologically active. ReNcell® VM may be used for a variety of research applications such as studies of neurotoxicity, neurogenesis, electrophysiology, neurotransmitter and receptor functions. ReNcell® VM cells have been obtained in a legal and ethical manner, compliant with current local informed consent procedures.
ReNcell Now Used in a 3D Cell Model of Alzheimer's Disease!
Tanzi, R and Kim, DY et. al. A three-dimensional human neural cell culture model of Alzheimer's disease. Nature. 2014 Nov 13;515(7526):274-8.
(Published in Nature)
|Materials Required but Not Delivered||1. ReNcell® NSC Maintenance Medium (Millipore Cat. No. SCM005)
2. ReNcell® NSC Freezing Medium (Millipore Cat. No. SCM007)
3. Basic fibroblast growth factor (bFGF; FGF-2; Specific Activity > 2 X 106 Units/mg. Millipore Cat. No. GF003)
4. Epidermal growth factor (EGF; Specific Activity > 1 x 107 Units/mg; MilliporeCat. No. GF001)
5. Laminin (Millipore Cat. No. CC095)
6. DMEM/F12 w/o HEPES, w/ L-Glutamine (Millipore Cat. No. DF-042-B)
7. Accutase (Millipore Cat. No. SCR005)
8. Tissue culture-ware
9. Phosphate-Buffered Saline (1X PBS) (Millipore Cat. No. BSS-1005-B)
10. Fixative (e.g. 4% Paraformaldehyde in 1X PBS)
11. Blocking Solution (5% normal donkey serum, 0.3% Triton X-100 in 1X PBS)
12. Primary and secondary antibodies
13. 4'-6-Diamidino-2-phenylindole (DAPI) / PBS solution
14. Anti-fading mounting solution (DABCO/PVA)
|Application||ReNcell® VM is an immortalized human neural progenitor cell line with the ability to readily differentiate into neurons & glial cells.|
|Source||Human ventral mesencephalon brain tissue|
|Stem Cell Type||
|Cell Line Type||
|Safety Information according to GHS|
|Product Usage Statements|
|Quality Assurance||Each lot of ReNcell® VM cells has been validated for high level of expression of Nestin and Sox 2 and for their self-renewal and multi-lineage differentiation capacities. Cells also display normal karyotype as assessed by chromosome spread and tested negative for mycoplasma. Maternal blood has been screened for HIV, HTLV, Hepatitis B and C.|
|Material Size||1 ea|
ReNcell® VM Human Neural Progenitor Cell LIne SDS
|Reference overview||Application||Pub Med ID|
|A dynamic view of the proteomic landscape during differentiation of ReNcell VM cells, an immortalized human neural progenitor line.|
Song, Y; Subramanian, K; Berberich, MJ; Rodriguez, S; Latorre, IJ; Luria, CM; Everley, R; Albers, MW; Mitchison, TJ; Sorger, PK
Sci Data 6 190016 2019
The immortalized human ReNcell VM cell line represents a reproducible and easy-to-propagate cell culture system for studying the differentiation of neural progenitors. To better characterize the starting line and its subsequent differentiation, we assessed protein and phospho-protein levels and cell morphology over a 15-day period during which ReNcell progenitors differentiated into neurons, astrocytes and oligodendrocytes. Five of the resulting datasets measured protein levels or states of phosphorylation based on tandem-mass-tag (TMT) mass spectrometry and four datasets characterized cellular phenotypes using high-content microscopy. Proteomic analysis revealed reproducible changes in pathways responsible for cytoskeletal rearrangement, cell phase transitions, neuronal migration, glial differentiation, neurotrophic signalling and extracellular matrix regulation. Proteomic and imaging data revealed accelerated differentiation in cells treated with the poly-selective CDK and GSK3 inhibitor kenpaullone or the HMG-CoA reductase inhibitor mevastatin, both of which have previously been reported to promote neural differentiation. These data provide in-depth information on the ReNcell progenitor state and on neural differentiation in the presence and absence of drugs, setting the stage for functional studies.
|A three-dimensional human neural cell culture model of Alzheimer's disease.|
Choi, SH; Kim, YH; Hebisch, M; Sliwinski, C; Lee, S; D'Avanzo, C; Chen, H; Hooli, B; Asselin, C; Muffat, J; Klee, JB; Zhang, C; Wainger, BJ; Peitz, M; Kovacs, DM; Woolf, CJ; Wagner, SL; Tanzi, RE; Kim, DY
Alzheimer's disease is the most common form of dementia, characterized by two pathological hallmarks: amyloid-β plaques and neurofibrillary tangles. The amyloid hypothesis of Alzheimer's disease posits that the excessive accumulation of amyloid-β peptide leads to neurofibrillary tangles composed of aggregated hyperphosphorylated tau. However, to date, no single disease model has serially linked these two pathological events using human neuronal cells. Mouse models with familial Alzheimer's disease (FAD) mutations exhibit amyloid-β-induced synaptic and memory deficits but they do not fully recapitulate other key pathological events of Alzheimer's disease, including distinct neurofibrillary tangle pathology. Human neurons derived from Alzheimer's disease patients have shown elevated levels of toxic amyloid-β species and phosphorylated tau but did not demonstrate amyloid-β plaques or neurofibrillary tangles. Here we report that FAD mutations in β-amyloid precursor protein and presenilin 1 are able to induce robust extracellular deposition of amyloid-β, including amyloid-β plaques, in a human neural stem-cell-derived three-dimensional (3D) culture system. More importantly, the 3D-differentiated neuronal cells expressing FAD mutations exhibited high levels of detergent-resistant, silver-positive aggregates of phosphorylated tau in the soma and neurites, as well as filamentous tau, as detected by immunoelectron microscopy. Inhibition of amyloid-β generation with β- or γ-secretase inhibitors not only decreased amyloid-β pathology, but also attenuated tauopathy. We also found that glycogen synthase kinase 3 (GSK3) regulated amyloid-β-mediated tau phosphorylation. We have successfully recapitulated amyloid-β and tau pathology in a single 3D human neural cell culture system. Our unique strategy for recapitulating Alzheimer's disease pathology in a 3D neural cell culture model should also serve to facilitate the development of more precise human neural cell models of other neurodegenerative disorders.
|Dynamic mass redistribution assay decodes differentiation of a neural progenitor stem cell.|
Pai, S; Verrier, F; Sun, H; Hu, H; Ferrie, AM; Eshraghi, A; Fang, Y
Journal of biomolecular screening 17 1180-91 2012
Stem cells hold great potential in drug discovery and development. However, challenges remain to quantitatively measure the functions of stem cells and their differentiated products. Here, we applied fluorescent imaging, quantitative real-time PCR, and label-free dynamic mass redistribution (DMR) assays to characterize the differentiation process of the ReNcell VM human neural progenitor stem cell. Immunofluorescence imaging showed that after growth factor withdrawal, the neuroprogenitor stem cell was differentiated into dopaminergic neurons, astrocytes, and oligodendrocytes, thus creating a neuronal cell system. High-performance liquid chromatography analysis showed that the differentiated cell system released dopamine upon depolarization with KCl. In conjunction with quantitative real-time PCR, DMR assays using a G-protein-coupled receptor agonist library revealed that a subset of receptors, including dopamine D(1) and D(4) receptors, underwent marked alterations in both receptor expression and signaling pathway during the differentiation process. These findings suggest that DMR assays can decode the differentiation process of stem cells at the cell system level.
|Interplay between REST and nucleolin transcription factors: a key mechanism in the overexpression of genes upon increased phosphorylation.|
Tediose, Teeo, et al.
Nucleic acids research, (2010) 2010
Non-malignant cells can be transformed via the activation of kinases that control degradation of neural-restrictive silencer factor (REST). Here, we identify a mechanism that contributes to the activation of genes, expression of which is controlled by responsive elements containing overlapping binding sites for REST and nucleolin. We demonstrate that both phosphorylated and non-phosphorylated nucleolin-bound DNA; however, only phosphorylated nucleolin successfully competed with either full-length REST or a REST-derived DNA-binding peptide, REST68, for binding to the overlapping binding sites. We show that this interplay between the two transcription factors regulates the activation of cell survival and immunomodulatory genes in tumors and non-malignant cells with activated protein kinase C, which is accompanied with alterations in cell proliferation and apoptosis. We propose a model for the regulation of these genes, which brings a new insight into the molecular mechanisms that control cellular transformation driven by activation of protein kinases.
|Transcriptional signature and memory retention of human-induced pluripotent stem cells.|
Marchetto, Maria C N, et al.
PLoS ONE, 4: e7076 (2009) 2009
Genetic reprogramming of somatic cells to a pluripotent state (induced pluripotent stem cells or iPSCs) by over-expression of specific genes has been accomplished using mouse and human cells. However, it is still unclear how similar human iPSCs are to human Embryonic Stem Cells (hESCs). Here, we describe the transcriptional profile of human iPSCs generated without viral vectors or genomic insertions, revealing that these cells are in general similar to hESCs but with significant differences. For the generation of human iPSCs without viral vectors or genomic insertions, pluripotent factors Oct4 and Nanog were cloned in episomal vectors and transfected into human fetal neural progenitor cells. The transient expression of these two factors, or from Oct4 alone, resulted in efficient generation of human iPSCs. The reprogramming strategy described here revealed a potential transcriptional signature for human iPSCs yet retaining the gene expression of donor cells in human reprogrammed cells free of viral and transgene interference. Moreover, the episomal reprogramming strategy represents a safe way to generate human iPSCs for clinical purposes and basic research.
|Differential development of neuronal physiological responsiveness in two human neural stem cell lines.|
Donato, R; Miljan, EA; Hines, SJ; Aouabdi, S; Pollock, K; Patel, S; Edwards, FA; Sinden, JD
BMC neuroscience 8 36 2007
Neural stem cells (NSCs) are powerful research tools for the design and discovery of new approaches to neurodegenerative disease. Overexpression of the myc family transcription factors in human primary cells from developing cortex and mesencephalon has produced two stable multipotential NSC lines (ReNcell VM and CX) that can be continuously expanded in monolayer culture.In the undifferentiated state, both ReNcell VM and CX are nestin positive and have resting membrane potentials of around -60 mV but do not display any voltage-activated conductances. As initially hypothesized, using standard methods (stdD) for differentiation, both cell lines can form neurons, astrocytes and oligodendrocytes according to immunohistological characteristics. However it became clear that this was not true for electrophysiological features which designate neurons, such as the firing of action potentials. We have thus developed a new differentiation protocol, designated 'pre-aggregation differentiation' (preD) which appears to favor development of electrophysiologically functional neurons and to lead to an increase in dopaminergic neurons in the ReNcell VM line. In contrast, the protocol used had little effect on the differentiation of ReNcell CX in which dopaminergic differentiation was not observed. Moreover, after a week of differentiation with the preD protocol, 100% of ReNcell VM featured TTX-sensitive Na+-channels and fired action potentials, compared to 25% after stdD. Currents via other voltage-gated channels did not appear to depend on the differentiation protocol. ReNcell CX did not display the same electrophysiological properties as the VM line, generating voltage-dependant K+ currents but no Na+ currents or action potentials under either stdD or preD differentiation.These data demonstrate that overexpression of myc in NSCs can be used to generate electrophysiologically active neurons in culture. Development of a functional neuronal phenotype may be dependent on parameters of isolation and differentiation of the cell lines, indicating that not all human NSCs are functionally equivalent.
|ReNcell VM Human Neural Progenitor Cell Line|
Newsletters / Publications
|Cellutions Newsletter: 2007, Volume 1|
|Cellutions Newsletter: 2008, Volume 1|
|Cellutions Newsletter: 2008, Volume 3|