NG2 and Olig2 Expression Provides Evidence for Phenotypic Deregulation of Cultured Central Nervous System and Peripheral Nervous System Neural Precursor Cells

Dromard, Cécile; Bartolami, Sylvain; Deleyrolle, Loic P.; Takebayashi, Hirohide; Ripoll, Camille; Simonneau, Lionel; Prome, Sylvie; Puech, Sylvie; Ba, Christophe Tran Van; Duperray, Christophe; Valmier, Jean; Privat, Alain; Hugnot, Jean‐Philippe

doi:10.1634/stemcells.2005-0556

Cited by 46 publications

(48 citation statements)

References 77 publications

(110 reference statements)

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“…Surprisingly, Olig2 transcripts were expressed in both neural tube and BC derivatives. However, the absence of Olig2 transcripts in meninges but their presence in BC derivatives indicated induction of Olig2 transcripts upon this culture condition, as previously reported (11).…”

Section: Resultssupporting

confidence: 87%

“…4). This suggests that BC-derived cells acquired CNS features or underwent deregulation in response to EGF/FGF2 treatment as previously reported for embryonic NPC and DRG stem cells (11). However, Olig2 induction in BC progeny was not sufficient to redirect BC into cells of the oligodendrocyte lineage, as nuclear Nkx2.2 was not detected, even in conditions favoring oligodendrocyte differentiation (NPC differentiation medium).…”

Section: Discussionsupporting

confidence: 68%

See 1 more Smart Citation

Boundary cap cells are peripheral nervous system stem cells that can be redirected into central nervous system lineages

Zujovic

Thibaud

Bachelin

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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Boundary cap cells (BC), which express the transcription factor Krox20, participate in the formation of the boundary between the central nervous system and the peripheral nervous system. To study BC stemness, we developed a method to purify and amplify BC in vitro from Krox20Cre/+ , R26RYFP/+ mouse embryos. We show that BC progeny are EGF/FGF2-responsive, form spheres, and express neural crest markers. Upon growth factor withdrawal, BC progeny gave rise to multiple neural crest and CNS lineages. Transplanted into the developing murine forebrain, they successfully survived, migrated, and integrated within the host environment. Surprisingly, BC progeny generated exclusively CNS cells, including neurons, astrocytes, and myelin-forming oligodendrocytes. In vitro experiments indicated that a sequential combination of ventralizing morphogens and glial growth factors was necessary to reprogram BC into oligodendrocytes. Thus, BC progeny are endowed with differentiation plasticity beyond the peripheral nervous system. The demonstration that CNS developmental cues can reprogram neural crest-derived stem cells into CNS derivatives suggests that BC could serve as a source of cell type-specific lineages, including oligodendrocytes, for cell-based therapies to treat CNS disorders.dysmyelination | reprogramming | transplantation B oundary cap cells (BC) are neural crest (NC) derivatives and were first described as discrete cell clusters localized at the dorsal root entry zone and motor exit point of the embryonic spinal cord (1). Their specific location at the central nervous system (CNS) and peripheral nervous system (PNS) interface and genetic ablation (2) showed that BC are involved in the formation of PNS-CNS boundaries. BC maintain spinal cord integrity by inhibiting motoneuron cell bodies exit to the periphery (3). Fate mapping, based on the exclusive expression of the zinc finger transcription factor Krox20 by BC between embryonic day (E)10.5 and E15.5, showed that migrating BC derivatives differentiate into Schwann cells (SC) in spinal roots, and satellite cells and a subset of nociceptive neurons in dorsal root ganglia (DRG), suggesting their multipotency (2). HjerlingLeffler and collaborators (4) highlighted the presence of a pluripotent population in the E11 mouse DRG capable of selfrenewal and differentiation into multiple NC derivatives. We reported that BC have a defined molecular signature intermediate between NC cells and SC precursors (3). Furthermore, when transplanted remotely from a focal myelin lesion of the spinal cord, BC generated remyelinating SC and few oligodendrocytes. Overall, these results strongly suggest that BC are stem cells of the embryonic PNS. However, the definition of BC stemness has been hampered by the difficulty of purifying and amplifying this restricted population. In this study, we used cellfate mapping, FACS, and microdissection to isolate, expand, and assess BC stemness in vitro and in vivo. Our data show that BC progeny behave as stem cells that can acquire differentiation plasti...

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Section: Resultssupporting

confidence: 87%

Section: Discussionsupporting

confidence: 68%

Boundary cap cells are peripheral nervous system stem cells that can be redirected into central nervous system lineages

Zujovic

Thibaud

Bachelin

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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“…NG2, PDGFR-a and Olig2, relatively specific markers for oligodendrocyte precursors, 26 were found by quantitative flow cytometry expressed in a similar proportion of cells (460%) and with similar intensity both in shCon and shATM neurospheres and at D0 (Supplementary Figure S5). The expression of these oligodendrocyte lineage markers in a large proportion of our neural stem cells is in agreement with the findings in murine neurospheres.…”

Section: Effects Of Atm Ablation In Human Neural Stem Cells L Carlessmentioning

confidence: 82%

“…The expression of these oligodendrocyte lineage markers in a large proportion of our neural stem cells is in agreement with the findings in murine neurospheres. 26 The flow cytometry analysis showed also a faint expression of CNPase, a marker of immature oligodendrocytes, 27 and virtual absence of GalC staining (Supplementary Figure S5). The regulation during differentiation of Olig2 and PDGFR-a assessed on western blots showed a modest reduction in Olig2 levels between D0 and D3 in shATM relative to shATM, and a marked drop in PDGFR-a levels at D10, more effective in shATM than shCon, particularly at D17 (Supplementary Figure S6).…”

Section: Effects Of Atm Ablation In Human Neural Stem Cells L Carlessmentioning

confidence: 98%

DNA-damage response, survival and differentiation in vitro of a human neural stem cell line in relation to ATM expression

et al. 2009

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Ataxia-telangiectasia (A-T) is a neurodegenerative disorder caused by defects in the ATM kinase, a component of the DNA-damage response (DDR). Here, we employed an immortalized human neural stem-cell line (ihNSC) capable of differentiating in vitro into neurons, oligodendrocytes and astrocytes to assess the ATM-dependent response and outcome of ATM ablation. The time-dependent differentiation of ihNSC was accompanied by an upregulation of ATM and DNA-PK, sharp downregulation of ATR and Chk1, transient induction of p53 and by the onset of apoptosis in a fraction of cells. The response to ionizing radiation (IR)-induced DNA lesions was normal, as attested by the phosphorylation of ATM and some of its substrates (e.g., Nbs1, Smc1, Chk2 and p53), and by the kinetics of c-H2AX nuclear foci formation. Depletion in these cells of ATM by shRNA interference (shATM) attenuated the differentiation-associated apoptosis and response to IR, but left unaffected the growth, self-renewal and genomic stability. shATM cells generated a normal number of MAP2/b-tubulin III þ neurons, but a reduced number of GalC þ oligodendrocytes, which were nevertheless more susceptible to oxidative stress. Altogether, these findings highlight the potential of ihNSCs as an in vitro model system to thoroughly assess, besides ATM, the role of DDR genes in neurogenesis and/or neurodegeneration.

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“…Therefore, we propose that the majority of the cells that are reprogrammed in our cultures are NCSCs-like cells. Importantly, we can exclude contamination of our cultures with spinal cord cells as (1) the dissection procedure is well established, simple, and avoids CNS contaminations (Dromard et al, 2007); (2) acutely isolated cells in the culture lack spinal cord marker expression (Nkx2.2, Nkx6.1, Pax6 ) ( Table 2) and express the DRG-specific Runx3 (Table 2); (3) the expression profile in early short-term cultures reflects PNS but not CNS progenitors (Fig. 1); and (4) spinal cord progenitors but not DRG-derived neurospheres can be maintained in medium containing EGF alone (data not shown).…”

Section: Identity Of Sphere-forming Cells In Embryonic Drgmentioning

confidence: 99%

Peripheral Nervous System Progenitors Can Be Reprogrammed to Produce Myelinating Oligodendrocytes and Repair Brain Lesions

Binder¹,

Rukavina²,

Hassani³

et al. 2011

J. Neurosci.

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Neural crest stem cells (NCSCs) give rise to the neurons and glia of the peripheral nervous system (PNS). NCSC-like cells can be isolated from multiple peripheral organs and maintained in neurosphere culture. Combining in vitro culture and transplantation, we show that expanded embryonic NCSC-like cells lose PNS traits and are reprogrammed to generate CNS cell types. When transplanted into the embryonic or adult mouse CNS, they differentiate predominantly into cells of the oligodendrocyte lineage without any signs of tumor formation. NCSC-derived oligodendrocytes generate CNS myelin and contribute to the repair of the myelin deficiency in shiverer mice. These results demonstrate a reprogramming of PNS progenitors to CNS fates without genetic modification and imply that PNS cells could be a potential source for cell-based CNS therapy.

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NG2 and Olig2 Expression Provides Evidence for Phenotypic Deregulation of Cultured Central Nervous System and Peripheral Nervous System Neural Precursor Cells

Abstract: STEM CELLS 2007;25:340 -353

Cited by 46 publications

References 77 publications

Boundary cap cells are peripheral nervous system stem cells that can be redirected into central nervous system lineages

Boundary cap cells are peripheral nervous system stem cells that can be redirected into central nervous system lineages

DNA-damage response, survival and differentiation in vitro of a human neural stem cell line in relation to ATM expression

Peripheral Nervous System Progenitors Can Be Reprogrammed to Produce Myelinating Oligodendrocytes and Repair Brain Lesions

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