Generation of iPS Cells from Human Umbilical Vein Endothelial Cells by Lentiviral Transduction and Their Differentiation to Neuronal Lineage

Shutova, Maria V.; Честков, И. В.; Bogomazova, Alexandra N.; Lagarkova, M. A.; Kiselev, Sergey L.

doi:10.1007/978-1-61779-267-0_11

Cited by 9 publications

(7 citation statements)

References 20 publications

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“…For the production of recombinant lentiviruses, a Phoenix packaging cell line was transfected with the lentiviral vectors encoding the reprogramming factors LeGOhOct3/ 4, LeGO-hSox2, LeGO-hKlf4, LeGO-hc-Myc as previously described [18]. …”

Section: Methodsmentioning

confidence: 99%

Patient-Specific Induced Pluripotent Stem Cells for SOD1-Associated Amyotrophic Lateral Sclerosis Pathogenesis Studies

et al. 2014

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The genetic reprogramming technology allows one to generate pluripotent stem cells for individual patients. These cells, called induced pluripotent stem cells (iPSCs), can be an unlimited source of specialized cell types for the body. Thus, autologous somatic cell replacement therapy becomes possible, as well as the generation of in vitro cell models for studying the mechanisms of disease pathogenesis and drug discovery. Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disorder that leads to a loss of upper and lower motor neurons. About 10% of cases are genetically inherited, and the most common familial form of ALS is associated with mutations in the SOD1 gene. We used the reprogramming technology to generate induced pluripotent stem cells with patients with familial ALS. Patient-specific iPS cells were obtained by both integration and transgene-free delivery methods of reprogramming transcription factors. These iPS cells have the properties of pluripotent cells and are capable of direct differentiation into motor neurons.

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Section: Methodsmentioning

confidence: 99%

Patient-Specific Induced Pluripotent Stem Cells for SOD1-Associated Amyotrophic Lateral Sclerosis Pathogenesis Studies

et al. 2014

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“…To obtain neural stem cells (NSCs) for subsequent differentiation into neuronal and glial lineages, hiPSCs were cultured in DMEM/F12 (Gibco, Waltham, MA, USA) supplemented with N-2 (to 1X, Gibco, Waltham, MA, USA), 2 mM L-glutamine (PanEco, Moscow, Russia), 100 mg/L penicillin-streptomycin (PanEco, Moscow, Russia), 10 µM SB431542 (Stemcell Technologies, Vancouver, BC, Canada), 2 µM dorsomorphin (Stemcell Technologies, Vancouver, BC, Canada) and 200 nM LDN193189 (Sigma-Aldrich, St. Louis, MO, USA) [ 48 ]. To obtain NPCs, NSCs were cultured for 2 weeks in DMEM/F12 (PanEco, Moscow, Russia) supplemented with B-27 (to 1X, Gibco, Waltham, MA, USA), 2 mM L-glutamine, 100 mg/L penicillin-streptomycin, 10 ng/mL FGF-2 (ProSpec, Ness-Ziona, Israel) and 1 µM purmorphamine (Stemcell Technologies, Vancouver, BC, Canada) [ 49 ]. To obtain GPCs, NSCs were cultured in a DMEM/F12-based growth medium supplemented with N-2, 2 mM L-glutamine, 100 mg/L penicillin-streptomycin and 1% fetal bovine serum (PanEco, Moscow, Russia).…”

Section: Methodsmentioning

confidence: 99%

“…[48]. To obtain NPCs, NSCs were cultured for 2 weeks in DMEM/F12 (PanEco, Moscow, Russia) supplemented with B-27 (to 1X, Gibco, Waltham, MA, USA), 2 mM L-glutamine, 100 mg/L penicillin-streptomycin, 10 ng/mL FGF-2 (ProSpec, Ness-Ziona, Israel) and 1 µM purmorphamine (Stemcell Technologies, Vancouver, BC, Canada) [49]. To obtain GPCs, NSCs were cultured in a DMEM/F12-based growth medium supplemented with N-2, 2 mM Lglutamine, 100 mg/L penicillin-streptomycin and 1% fetal bovine serum (PanEco, Moscow, Russia).…”

Section: Experimental Designmentioning

confidence: 99%

Therapeutic Effects of hiPSC-Derived Glial and Neuronal Progenitor Cells-Conditioned Medium in Experimental Ischemic Stroke in Rats

Salikhova

Бухарова

Cherkashova

et al. 2021

IJMS

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Transplantation of various types of stem cells as a possible therapy for stroke has been tested for years, and the results are promising. Recent investigations have shown that the administration of the conditioned media obtained after stem cell cultivation can also be effective in the therapy of the central nervous system pathology (hypothesis of their paracrine action). The aim of this study was to evaluate the therapeutic effects of the conditioned medium of hiPSC-derived glial and neuronal progenitor cells in the rat middle cerebral artery occlusion model of the ischemic stroke. Secretory activity of the cultured neuronal and glial progenitor cells was evaluated by proteomic and immunosorbent-based approaches. Therapeutic effects were assessed by overall survival, neurologic deficit and infarct volume dynamics, as well as by the end-point values of the apoptosis- and inflammation-related gene expression levels, the extent of microglia/macrophage infiltration and the numbers of formed blood vessels in the affected area of the brain. As a result, 31% of the protein species discovered in glial progenitor cells-conditioned medium and 45% in neuronal progenitor cells-conditioned medium were cell type specific. The glial progenitor cell-conditioned media showed a higher content of neurotrophins (BDNF, GDNF, CNTF and NGF). We showed that intra-arterial administration of glial progenitor cells-conditioned medium promoted a faster decrease in neurological deficit compared to the control group, reduced microglia/macrophage infiltration, reduced expression of pro-apoptotic gene Bax and pro-inflammatory cytokine gene Tnf, increased expression of anti-inflammatory cytokine genes (Il4, Il10, Il13) and promoted the formation of blood vessels within the damaged area. None of these effects were exerted by the neuronal progenitor cell-conditioned media. The results indicate pronounced cytoprotective, anti-inflammatory and angiogenic properties of soluble factors secreted by glial progenitor cells.

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“…To obtain NPСs, NSCs were cultured for 2 weeks in DMEM/F12 (PanEco) supplemented with B-27 (to 1X, Gibco), 2 mM L-glutamine, 100 mg/L penicillin-streptomycin, 10 ng/mL FGF-2 (ProSpec, UK) and 1 µM purmorphamine (Stemcell Technologies) [11].…”

Section: Methodsmentioning

confidence: 99%

Secretome-Mediated Neuroprotective Effects of the hiPSC-derived Glial and Neuronal Progenitor Cells in the Hypoxia-Induced Neuronal Damage (Comparative Study)

Salikhova

Бухарова

Cherkashova³

et al. 2021

Preprint

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Background: Stem cell secretomes hold great promise for regenerative medicine. This study is focused on the secretome-mediated neuroprotective effects of the human induced pluripotent stem cell-derived neuronal and glial progenitor cells. Therapeutic properties of the secretomes were assessed under conditions of the hypoxia-induced neuronal damage in vitro and in vivo. Methods: Secretory activity of the cultured neuronal and glial progenitor cells was analyzed by proteomic and immunosorbent-based approaches. Conditioned media collected from the cultures was tested for neuroprotective properties in vitro and in vivo.In vitro experiments involved exposure of SH-SY5Y cells to the conditioned media during the recovery from the cobalt chloride-induced hypoxia. Neuroprotective effects were assessed by cell survival and neurite outgrowth. Cell survival indicators included MTT and LDH tests, vital staining with propidium iodide and Hoechst 33342, and polymerase chain reaction assay for the expression of apoptosis-related genes. Neurite outgrowth was assessed by alterations in SH-SY5Y cell morphology and MAP2/GAP43 gene expression dynamics. In vivo experiments involved intra-arterial administration of the conditioned media to laboratory rats during the recovery from experimental ischemic stroke. Neuroprotective effects were assessed by overall survival, neurologic deficit and infarct volume dynamics, as well as by the end-point values of the apoptosis- and inflammation-related gene expression levels, the extent of microglia/macrophage infiltration, and the numbers of newly formed blood vessels in the affected area of the brain. Results: Secretomes of glial and neuronal progenitor cells partially overlapped, with specific proteins (found in secretome of one of the studied cultures and absent from the other) constituting, respectively, 31% and 45%. The glial progenitor cell-conditioned media showed higher content of neurotrophins (BDNF, GDNF, CNTF and NGF).Moreover, the glial progenitor cell-conditioned media was superior to the neuronal progenitor cell-conditioned media in facilitating neurite outgrowth and increasing SHSY-5Y cell survival after the cobalt dichloride-induced hypoxia. In addition, intra-arterial infusion of the glial progenitor cell-conditioned media to the animals after experimental ischemic stroke significantly enhanced functional recovery and promoted tissue repair at the site of brain damage, as indicated by reduced microglia/macrophage infiltration, decreased expression of pro-apoptotic gene Bax and pro-inflammatory cytokine gene Tnf, increased expression of anti-inflammatory cytokine genes (Il4, Il10, Il13), and increased numbers of newly formed blood vessels within the damaged area. None of these effects were exerted by the neuronal progenitor cell-conditioned media. Conclusions: The results indicate pronounced cytoprotective, anti-inflammatory and angionenic properties of soluble factors secreted by glial progenitor cells.

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Generation of iPS Cells from Human Umbilical Vein Endothelial Cells by Lentiviral Transduction and Their Differentiation to Neuronal Lineage

Cited by 9 publications

References 20 publications

Patient-Specific Induced Pluripotent Stem Cells for SOD1-Associated Amyotrophic Lateral Sclerosis Pathogenesis Studies

Patient-Specific Induced Pluripotent Stem Cells for SOD1-Associated Amyotrophic Lateral Sclerosis Pathogenesis Studies

Therapeutic Effects of hiPSC-Derived Glial and Neuronal Progenitor Cells-Conditioned Medium in Experimental Ischemic Stroke in Rats

Secretome-Mediated Neuroprotective Effects of the hiPSC-derived Glial and Neuronal Progenitor Cells in the Hypoxia-Induced Neuronal Damage (Comparative Study)

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