Engraftment of oligodendrocyte progenitor cells (OPCs), which form myelinating oligodendrocytes, has the potential to treat demyelinating diseases such as multiple sclerosis. However, conventional strategies for generating oligodendrocytes have mainly focused on direct differentiation into forebrain- or spinal cord–restricted oligodendrocytes without establishing or amplifying stem/progenitor cells. Taking advantage of a recently established culture system, we generated expandable EN1- and GBX2-positive glial-restricted progenitor-like cells (GPLCs) near the anterior hindbrain. These cells expressed PDGFRα, CD9, S100β, and SOX10 and mostly differentiated into GFAP-positive astrocytes and MBP-positive oligodendrocytes. RNA-seq analysis revealed that the transcriptome of GPLCs was similar to that of O4-positive OPCs, but distinct from that of rosette-type neural stem cells. Notably, engrafted GPLCs not only differentiated into GFAP-positive astrocytes but also myelinated the brains of adult shiverer mice 8 weeks after transplantation. Our strategy for establishing anterior hindbrain-specific GPLCs with gliogenic potency will facilitate their use in the treatment of demyelinating diseases and studies of the molecular mechanisms underlying glial development in the hindbrain.
Get full access to this article
View all access options for this article.
References
1.
ContiL and CattaneoE. (2010). Neural stem cell systems: physiological players or in vitro entities?. Nat Rev Neurosci, 11:176–187.
2.
KokovayE, ShenQ and TempleS. (2008). The incredible elastic brain: how neural stem cells expand our minds. Neuron, 60:420–429.
3.
RowitchDH. (2004). Glial specification in the vertebrate neural tube. Nat Rev Neurosci, 5:409–419.
4.
GageFH and TempleS. (2013). Neural stem cells: generating and regenerating the brain. Neuron, 80:588–601.
5.
RossiF and CattaneoE. (2002). Neural stem cell therapy for neurological diseases: dreams and reality. Nat Rev Neurosci, 3:401.
6.
TangY, YuP and ChengL. (2017). Current progress in the derivation and therapeutic application of neural stem cells. Cell Death Dis, 8:e3108.
7.
TrounsonA and McDonaldC. (2015). Stem cell therapies in clinical trials: progress and challenges. Cell Stem Cell, 17:11–22.
CasarosaS, BozziY and ContiL. (2014). Neural stem cells: ready for therapeutic applications?. Mol Cell Ther, 2:31–31.
10.
GoldmanSA. (2011). Progenitor cell-based treatment of the pediatric myelin disorders. Arch Neurol, 68:848–856.
11.
BayraktarOA, FuentealbaLC, Alvarez-BuyllaA and RowitchDH. Astrocyte development and heterogeneity. Cold Spring Harbor Perspectives Biol, 7:a020362-a020362.
12.
MatyashV and KettenmannH. (2010). Heterogeneity in astrocyte morphology and physiology. Brain Res Rev, 63:2–10.
13.
HochstimC, DeneenB, LukaszewiczA, ZhouQ and AndersonDJ. (2008). Identification of positionally distinct astrocyte subtypes whose identities are specified by a homeodomain code. Cell, 133:510–522.
14.
Kuzdas-WoodD, StefanovaN, JellingerKA, SeppiK, SchlossmacherMG, PoeweW and WenningGK. (2014). Towards translational therapies for multiple system atrophy. Prog Neurobiol, 118:19–35.
15.
ValeraE and MasliahE. (2018). The neuropathology of multiple system atrophy and its therapeutic implications. Auton Neurosci, 211:1–6.
16.
MolofskyAV, KrencikR, UllianEM, TsaiH-H, DeneenB, RichardsonWD, BarresBA and RowitchDH. (2012). Astrocytes and disease: a neurodevelopmental perspective. Genes Dev, 26:891–907.
17.
MungenastAE, SiegertS and TsaiLH. (2016). Modeling Alzheimer's disease with human induced pluripotent stem (iPS) cells. Mol Cell Neurosci, 73:13–31.
18.
AbatiE, Di FonzoA and CortiS. (2018). In vitro models of multiple system atrophy from primary cells to induced pluripotent stem cells. J Cell Mol Med, 22:2536–2546.
19.
KirkebyA, GrealishS, WolfDA, NelanderJ, WoodJ, LundbladM, LindvallO and ParmarM. (2012). Generation of regionally specified neural progenitors and functional neurons from human embryonic stem cells under defined conditions. Cell Rep, 1:703–714.
20.
LuJ, ZhongX, LiuH, HaoL, HuangCT, SherafatMA, JonesJ, AyalaM, LiL and ZhangSC. (2016). Generation of serotonin neurons from human pluripotent stem cells. Nat Biotechnol, 34:89–94.
21.
XiJ, LiuY, LiuH, ChenH, EmborgME and ZhangSC. (2012). Specification of midbrain dopamine neurons from primate pluripotent stem cells. Stem Cells (Dayton, Ohio), 30:1655–1663.
22.
KrencikR, WeickJP, LiuY, ZhangZJ and ZhangSC. (2011). Specification of transplantable astroglial subtypes from human pluripotent stem cells. Nat Biotechnol, 29:528–534.
23.
LiW, SunW, ZhangY, WeiW, AmbasudhanR, XiaP, TalantovaM, LinT, KimJ, et al. (2011). Rapid induction and long-term self-renewal of primitive neural precursors from human embryonic stem cells by small molecule inhibitors. Proc Natl Acad Sci U S A, 108:8299–8304.
24.
LuJ, LiuH, HuangCT, ChenH, DuZ, LiuY, SherafatMA and ZhangSC. (2013). Generation of integration-free and region-specific neural progenitors from primate fibroblasts. Cell Rep, 3:1580–1591.
25.
KochP, OpitzT, SteinbeckJA, LadewigJ and BrustleO. (2009). A rosette-type, self-renewing human ES cell-derived neural stem cell with potential for in vitro instruction and synaptic integration. Proc Natl Acad Sci U S A, 106:3225–3230.
26.
GorrisR, FischerJ, ErwesKL, KesavanJ, PetersonDA, AlexanderM, NothenMM, PeitzM, QuandelT, KarusM and BrustleO. (2015). Pluripotent stem cell-derived radial glia-like cells as stable intermediate for efficient generation of human oligodendrocytes. Glia, 63:2152–2167.
27.
GoldmanSA, NedergaardM and WindremMS. (2012). Glial progenitor cell-based treatment and modeling of neurological disease. Science, 338:491–495.
28.
WindremMS, OsipovitchM, LiuZ, BatesJ, Chandler-MilitelloD, ZouL, MunirJ, SchanzS, McCoyK, et al. (2017). Human iPSC glial mouse chimeras reveal glial contributions to schizophrenia. Cell Stem Cell, 21:195–208e6.
29.
WangS, BatesJ, LiX, SchanzS, Chandler-MilitelloD, LevineC, MaheraliN, StuderL, HochedlingerK, WindremM and GoldmanSA. (2013). Human iPSC-derived oligodendrocyte progenitor cells can myelinate and rescue a mouse model of congenital hypomyelination. Cell Stem Cell, 12:252–264.
30.
TanRH, KrilJJ, McGinleyC, HassaniM, Masuda-SuzukakeM, HasegawaM, MitoR, KiernanMC and HallidayGM. (2016). Cerebellar neuronal loss in amyotrophic lateral sclerosis cases with ATXN2 intermediate repeat expansions. Ann Neurol, 79:295–305.
31.
KoiralaS and CorfasG. (2010). Identification of novel glial genes by single-cell transcriptional profiling of Bergmann glial cells from mouse cerebellum. PLoS One, 5:e9198.
32.
HuBY, DuZW, LiXJ, AyalaM and ZhangSC. (2009). Human oligodendrocytes from embryonic stem cells: conserved SHH signaling networks and divergent FGF effects. Development, 136:1443–1452.
33.
DouvarasP, WangJ, ZimmerM, HanchukS, O'BaraMA, SadiqS, SimFJ, GoldmanJ and FossatiV. (2014). Efficient generation of myelinating oligodendrocytes from primary progressive multiple sclerosis patients by induced pluripotent stem cells. Stem Cell Rep, 3:250–259.
34.
PiaoJ, MajorT, AuyeungG, PolicarpioE, MenonJ, DromsL, GutinP, UryuK, TchieuJ, SouletD and TabarV. (2015). Human embryonic stem cell-derived oligodendrocyte progenitors remyelinate the brain and rescue behavioral deficits following radiation. Cell Stem Cell, 16:198–210.
35.
HashimotoR, HoriK, OwaT, MiyashitaS, DewaK, MasuyamaN, SakaiK, HayaseY, SetoY, et al. (2016). Origins of oligodendrocytes in the cerebellum, whose development is controlled by the transcription factor, Sox9. Mech Dev, 140:25–40.
36.
SgaierSK, MilletS, VillanuevaMP, BerenshteynF, SongC and JoynerAL. (2005). Morphogenetic and cellular movements that shape the mouse cerebellum: insights from genetic fate mapping. Neuron, 45:27–40.
37.
TailorJ, KittappaR, LetoK, GatesM, BorelM, PaulsenO, SpitzerS, KaradottirRT, RossiF, FalkA and SmithA. (2013). Stem cells expanded from the human embryonic hindbrain stably retain regional specification and high neurogenic potency. J Neurosci, 33:12407–12422.
38.
KangPJ, ZhengJ, LeeG, SonD, KimIY, SongG, ParkG and YouS. (2019). Glycine decarboxylase regulates the maintenance and induction of pluripotency via metabolic control. Metab Eng, 53:35–47.
39.
IrvingC and MasonI. (2000). Signalling by FGF8 from the isthmus patterns anterior hindbrain and establishes the anterior limit of Hox gene expression. Development, 127:177–186.
40.
WaiteMR, SkaggsK, KavianyP, SkidmoreJM, CauseretF, MartinJF and MartinDM. (2012). Distinct populations of GABAergic neurons in mouse rhombomere 1 express but do not require the homeodomain transcription factor PITX2. Mol Cell Neurosci, 49:32–43.
41.
PintoL and GotzM. (2007). Radial glial cell heterogeneity—the source of diverse progeny in the CNS. Prog Neurobiol, 83:2–23.
42.
HowardBM, ZhichengM, FilipovicR, MooreAR, AnticSD and ZecevicN. (2008). Radial glia cells in the developing human brain. Neuroscientist, 14:459–473.
43.
MalatestaP, AppolloniI and CalzolariF. (2008). Radial glia and neural stem cells. Cell Tissue Res, 331:165–178.
44.
Turrero Garcia M and HarwellCC. (2017). Radial glia in the ventral telencephalon. FEBS Lett, 591:3942–3959.
45.
DouvarasP and FossatiV. (2015). Generation and isolation of oligodendrocyte progenitor cells from human pluripotent stem cells. Nat Protoc, 10:1143–1154.
46.
ThakurelaS, GardingA, JungRB, MullerC, GoebbelsS, WhiteR, WernerHB and TiwariVK. (2016). The transcriptome of mouse central nervous system myelin. Sci Rep, 6:25828.
47.
LuQR, YukD, AlbertaJA, ZhuZ, PawlitzkyI, ChanJ, McMahonAP, StilesCD and RowitchDH. (2000). Sonic hedgehog—regulated oligodendrocyte lineage genes encoding bHLH proteins in the mammalian central nervous system. Neuron, 25:317–329.
48.
OrentasDM, HayesJE, DyerKL and MillerRH. (1999). Sonic hedgehog signaling is required during the appearance of spinal cord oligodendrocyte precursors. Development, 126:2419–2429.
49.
ZhouQ and AndersonDJ. (2002). The bHLH transcription factors OLIG2 and OLIG1 couple neuronal and glial subtype specification. Cell, 109:61–73.
50.
StacpooleSR, SpitzerS, BilicanB, CompstonA, KaradottirR, ChandranS and FranklinRJ. (2013). High yields of oligodendrocyte lineage cells from human embryonic stem cells at physiological oxygen tensions for evaluation of translational biology. Stem Cell Rep, 1:437–450.
51.
FurushoM, KagaY, IshiiA, HebertJM and BansalR. (2011). Fibroblast growth factor signaling is required for the generation of oligodendrocyte progenitors from the embryonic forebrain. J Neurosci, 31:5055–5066.
52.
NeryS, WichterleH and FishellG. (2001). Sonic hedgehog contributes to oligodendrocyte specification in the mammalian forebrain. Development, 128:527–540.
53.
KessarisN, FogartyM, IannarelliP, GristM, WegnerM and RichardsonWD. (2006). Competing waves of oligodendrocytes in the forebrain and postnatal elimination of an embryonic lineage. Nat Neurosci, 9:173–179.
BlackburnD, SargsyanS, MonkPN and ShawPJ. (2009). Astrocyte function and role in motor neuron disease: a future therapeutic target?. Glia, 57:1251–1264.
56.
MendoncaLS, NobregaC, HiraiH, KasparBK and Pereira de AlmeidaL. (2015). Transplantation of cerebellar neural stem cells improves motor coordination and neuropathology in Machado-Joseph disease mice. Brain, 138:320–335.
57.
PolSU, PolancoJJ, SeidmanRA, O'BaraMA, ShayyaHJ, DietzKC and SimFJ. (2017). Network-based genomic analysis of human oligodendrocyte progenitor differentiation. Stem Cell Rep, 9:710–723.
58.
RosenbergAB, RocoCM, MuscatRA, KuchinaA, SampleP, YaoZ, GrayL, PeelerDJ, MukherjeeS, et al. (2018). Single-cell profiling of the developing mouse brain and spinal cord with split-pool barcoding. Science, 360:176–182.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
