Human Astrocytes Derived from Glial Restricted Progenitors Support Regeneration of the Injured Spinal Cord

Haas, Christopher; Fischer, Itzhak

doi:10.1089/neu.2013.2915

Cited by 62 publications

(58 citation statements)

References 75 publications

(221 reference statements)

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“…The percentage of low conductance cells was the highest in VM-Ast cultures at DIV14 and decreased during maturation in VM at DIV35, while the percentages were similar in Ctx DIV14 and Ctx DIV35 (Supplemental Figure 1F). Of note, it has been shown that transplanted immature -but not mature -astrocytes exert neurotrophic support in the injured CNS (25)(26)(27)(28)(29). In addition to a wound-healing capacity superior to that in astrocytes at DIV33-35 (Supplemental Figure 1G) in a scratch injury model (28), astrocytes at DIV12-14 showed greater capacity growing on high levels of chondroitin sulfate proteoglycans (CSPGs) (Supplemental Figure 1H) in an in vitro model recreating the strongly inhibitory CSPG gradient observed in the glial scar in vivo (25).…”

Section: Resultsmentioning

confidence: 99%

Cografting astrocytes improves cell therapeutic outcomes in a Parkinson’s disease model

Song¹,

Oh²,

Kwon³

et al. 2017

Journal of Clinical Investigation

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In this study, we have attempted to exploit the neurotrophic actions of astrocytes and Nurr1+Foxa2 functions in this cell type to improve the therapeutic outcomes of NPC transplantation using an animal model of PD. Our in vitro assays using a coculture system and conditioned media treatment revealed that astrocytes, especially those cultured from the ventral midbrain (VM), where HNF3β) is a potent cofactor that synergizes the Nurr1-mediated antiinflammatory roles in glia (16). Furthermore, forced coexpression of Nurr1 and Foxa2 (Nurr1+Foxa2) in glia promotes the synthesis and release of various neurotrophic factors, indicating that engineering of Nurr1 and Foxa2 in glia could become a powerful strategy for treating CNS disorders. Representative images of TH + DA neurons differentiated from VM-NPCs in the presence of Ctx-NPCs (control), Ctx-Ast, or VM-Ast. Scale bar: 100 μm. Insets, enlarged images of the boxed areas (original magnification, ×400). (C) DA neuronal yields at differentiation day 6 (D6). (D-H) Morphometric measurement of DA neuron maturity assessed by neurite outgrowth lengths (D), soma size (E), and by the Sholl test (F and G). In the Sholl analysis, the total number of neurite crossings was counted in each circle with the radius increasing in steps of 15 μm (F). The critical value (G) is the radius at which there was a maximum number of neurite crossings. *P < 0.05, significantly different from the control; # P < 0.05, significantly different from Ctx-Ast, 1-way ANOVA. n = 3-5 wells (C) and 100 (D and E) and 25-30 (F and G) TH + cells in each group.(H) Representative TH + neuronal morphologies reconstructed in 3D by Neurolucida. (I-L) Synaptic densities on TH + fibers. (I) Representative confocal images of synapsin + TH + fibers. Scale bar: 25 μm. Puncta positive for the synaptic vesicle-specific markers SV2 (J), synapsin (K), and bassoon (L) on TH + fibers were counted. *P < 0.05, significantly different from the control; # P < 0.05, significantly different from Ctx-Ast. n = 20 TH + fibers for each group. (M) Presynaptic DA release. n = 3 independent cultures. DA levels were measured in the media conditioned in the differentiated cultures for 24 hours (D13-D15) and in cultures evoked by KCl-mediated depolarization for 30 minutes. *P < 0.05; # P < 0.05, 1-way ANOVA. The Journal of Clinical Investigation R E S E A R C H A R T I C L E4 6 5 jci.org Volume 128 Number 1 January 2018 (Supplemental Figure 1B; supplemental material available online with this article; https://doi.org/10.1172/JCI93924DS1), immunocytochemical analyses revealed that major cell populations in the Ctx-and VM-astroglial cultures at day in vitro (DIV) 14 were positive for GFAP (85% and 82%), CD44 (79% and 76 %), GLT-1 (58% and 71 %), and GLAST protein (71% and 77%), respectively (Supplemental Figure 1A), except S100β (20% and 32%), a soluble protein associated with harmful reactivation of astrocytes (22). A few (0.24% ± 0.35% in Ctx-Ast and 0.43% ± 0.39% in VM-Ast, where Ast indicates astrocytes) and none of the cells in these c...

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Section: Resultsmentioning

confidence: 99%

Cografting astrocytes improves cell therapeutic outcomes in a Parkinson’s disease model

Song¹,

Oh²,

Kwon³

et al. 2017

Journal of Clinical Investigation

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“…Specifically, with respect to GFAP, previous studies have shown heterogeneity in both maturity and marker expression in cultures of human stem cell-derived astrocytes, but identify GFAP expression as a marker for astroglial identity (90), and GFAP is commonly used to characterize astroglial differentiation in hNSC cultures (91)(92)(93). Critically, more mature astroglial markers such as GLAST and ALDH1L1 are absent in human cell cultures ,4 wk old (90).…”

Section: Discussionmentioning

confidence: 99%

Neutrophils Induce Astroglial Differentiation and Migration of Human Neural Stem Cells via C1q and C3a Synthesis

Hooshmand

Nguyen

Piltti

et al. 2017

The Journal of Immunology

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Inflammatory processes play a key role in pathophysiology of many neurologic diseases/trauma, but the effect of immune cells and factors on neurotransplantation strategies remains unclear. We hypothesized that cellular and humoral components of innate immunity alter fate and migration of human neural stem cells (hNSC). In these experiments, conditioned media collected from polymorphonuclear leukocytes (PMN) selectively increased hNSC astrogliogenesis and promoted cell migration in vitro. PMN were shown to generate C1q and C3a; exposure of hNSC to PMN-synthesized concentrations of these complement proteins promoted astrogliogenesis and cell migration. Furthermore, in vitro, Abs directed against C1q and C3a reversed the fate and migration effects observed. In a proof-of-concept in vivo experiment, blockade of C1q and C3a transiently altered hNSC migration and reversed astroglial fate after spinal cord injury. Collectively, these data suggest that modulation of the innate/humoral inflammatory microenvironment may impact the potential of cell-based therapies for recovery and repair following CNS pathology.

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“…Such progenitors can be isolated from embryonic tissue and expanded for therapeutic applications to reduce inhibition and provide a permissive environment for neuroprotection and axonal growth (Mothe and Tator, 2013; Okano et al, 2007; Tetzlaff et al, 2011). It is therefore critical to establish efficient culture protocols for large-scale cell preparation, and characterize these cell populations in terms of their phenotypic and functional properties under different environments (Haas et al, 2012; Haas and Fischer, 2013). The present study addresses these issues by analyzing the properties of glial progenitors after long term culturing and following exposure to inflammatory mediators.…”

Section: Discussionmentioning

confidence: 99%

Glial restricted precursors maintain their permissive properties after long-term expansion but not following exposure to pro-inflammatory factors

et al. 2015

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Glial restricted precursors (GRP) are a promising cellular source for transplantation therapy of spinal cord injury (SCI), capable of creating a permissive environment for axonal growth and regeneration. However, there are several issues regarding the nature of their permissive properties that remain unexplored. For example, cellular transplantation strategies for spinal cord repair require the preparation of a large number of cells, but it is unknown whether the permissive properties of GRP are maintained following the process of in vitro expansion. We used rat GRP isolated from the embryonic day 13.5 spinal cord to compare the properties of early (10-20 days) and late (120-140 days) passage GRP. We found that late passage GRP showed comparable effects on neurite outgrowth of adult rat DRG to early passage GRP in both in vitro co-culture and conditioned medium experiments. In addition, to further examine the effects of the inflammatory cascade activated in the aftermath of SCI on the microenvironment, we studied the direct effects of strong inflammatory mediators, Lipopolysaccharide and interferon gamma (LPS and IFNɤ, respectively), on the properties of GRP. We showed that exposure to these pro-inflammatory mediators altered GRP phenotype and attenuated their growth-promoting effects on neurite outgrowth in a dose dependent manner. Taken together, our data suggest that GRP maintain their growth-promoting properties following extensive in vitro passaging and underscore the importance of modulating the inflammatory environment at the injured spinal cord.

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Human Astrocytes Derived from Glial Restricted Progenitors Support Regeneration of the Injured Spinal Cord

Cited by 62 publications

References 75 publications

Cografting astrocytes improves cell therapeutic outcomes in a Parkinson’s disease model

Cografting astrocytes improves cell therapeutic outcomes in a Parkinson’s disease model

Neutrophils Induce Astroglial Differentiation and Migration of Human Neural Stem Cells via C1q and C3a Synthesis

Glial restricted precursors maintain their permissive properties after long-term expansion but not following exposure to pro-inflammatory factors

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