There is compelling evidence that the mature central nervous system (CNS) harbors stem cell populations outside conventional neurogenic regions. We previously demonstrated that brain pericytes (PCs) in both mouse and human exhibit multipotency to differentiate into various neural lineages following cerebral ischemia. PCs are found throughout the CNS, including cerebellum, but it remains unclear whether cerebellar PCs also form ischemia-induced multipotent stem cells (iSCs). In this study, we demonstrate that putative iSCs can be isolated from poststroke human cerebellum (cerebellar iSCs [cl-iSCs]). These cl-iSCs exhibited multipotency and differentiated into electrophysiologically active neurons. Neurogenic potential was also confirmed in single-cell suspensions. DNA microarray analysis revealed highly similar gene expression patterns between PCs and cl-iSCs, suggesting PC origin. Global gene expression comparison with cerebral iSCs revealed general similarity, but cl-iSCs differentially expressed certain cerebellum-specific genes. Thus, putative iSCs are present in poststroke cerebellum and possess region-specific traits, suggesting potential capacity to regenerate functional cerebellar neurons following ischemic stroke.
Get full access to this article
View all access options for this article.
References
1.
MokinM, Kass-HoutT, Kass-HoutO, DumontTM, KanP, SnyderKV, HopkinsLN, SiddiquiAH and LevyEI. (2012). Intravenous thrombolysis and endovascular therapy for acute ischemic stroke with internal carotid artery occlusion: a systematic review of clinical outcomes. Stroke, 43:2362–2368.
2.
ZaidatOO, SuarezJI, SunshineJL, TarrRW, AlexanderMJ, SmithTP, EnterlineDS, SelmanWR and LandisDM. (2005). Thrombolytic therapy of acute ischemic stroke: correlation of angiographic recanalization with clinical outcome. AJNR Am J Neuroradiol, 26:880–884.
3.
GoyalM, MenonBK, van ZwamWH, DippelDW, MitchellPJ, DemchukAM, DavalosA, MajoieCB, van der LugtA, (2016). Endovascular thrombectomy after large-vessel ischaemic stroke: a meta-analysis of individual patient data from five randomised trials. Lancet, 387:1723–1731.
4.
SalewskiRP, MitchellRA, LiL, ShenC, MilekovskaiaM, NagyA and FehlingsMG. (2015). Transplantation of induced pluripotent stem cell-derived neural stem cells mediate functional recovery following thoracic spinal cord injury through remyelination of axons. Stem Cells Transl Med, 4:743–754.
5.
KimuraH, YoshikawaM, MatsudaR, ToriumiH, NishimuraF, HirabayashiH, NakaseH, KawaguchiS, IshizakaS and SakakiT. (2005). Transplantation of embryonic stem cell-derived neural stem cells for spinal cord injury in adult mice. Neurol Res, 27:812–819.
6.
HonmaT, HonmouO, IihoshiS, HaradaK, HoukinK, HamadaH and KocsisJD. (2006). Intravenous infusion of immortalized human mesenchymal stem cells protects against injury in a cerebral ischemia model in adult rat. Exp Neurol, 199:56–66.
7.
AppaixF, NissouMF, van der SandenB, DreyfusM, BergerF, IssartelJP and WionD. (2014). Brain mesenchymal stem cells: the other stem cells of the brain?. World J Stem Cells, 6:134–143.
8.
LuP, BleschA and TuszynskiMH. (2004). Induction of bone marrow stromal cells to neurons: differentiation, transdifferentiation, or artifact?. J Neurosci Res, 77:174–191.
9.
NakagomiT, KuboS, Nakano-DoiA, SakumaR, LuS, NaritaA, KawaharaM, TaguchiA and MatsuyamaT. (2015). Brain vascular pericytes following ischemia have multipotential stem cell activity to differntiate into neural and vascular lineage cells. Stem Cells, 33:1962–1974.
10.
NakagomiT, MolnarZ, Nakano-DoiA, TaguchiA, SainoO, KuboS, ClausenM, YoshikawaH, NakagomiN and MatsuyamaT. (2011). Ischemia-induced neural stem/progenitor cells in the pia mater following cortical infarction. Stem Cells Dev, 20:2037–2051.
11.
SakumaR, KawaharaM, Nakano-DoiA, TakahashiA, TanakaY, NaritaA, Kuwahara-OtaniS, HayakawaT, YagiH, MatsuyamaT and NakagomiT. (2016). Brain pericytes serve as microglia-generating multipotent vascular stem cells following ischemic stroke. J Neuroinflammation, 13:57.
12.
GouveiaA, SeegobinM, KannangaraTS, HeL, WondisfordF, CominCH, CostaLDF, BeiqueJC, LagaceDC, LacosteB and WangJ. (2017). The aPKC-CBP pathway regulates post-stroke neurovascular remodeling and functional recovery. Stem Cell Rep, 9:1735–1744.
13.
SakumaR, TakahashiA, Nakano-DoiA, SawadaR, KamachiS, BeppuM, TakagiT, YoshimuraS, MatsuyamaT and NakagomiT. (2018). Comparative characterization of ischemia-induced brain multipotent stem cells with mesenchymal stem cells: similarities and differences. Stem Cells Dev, 27:1322–1338.
14.
TatebayashiK, TanakaY, Nakano-DoiA, SakumaR, KamachiS, ShirakawaM, UchidaK, KageyamaH, TakagiT, et al. (2017). Identification of multipotent stem cells in human brain tissue following stroke. Stem Cells Dev, 26:787–797.
15.
BastianAJ, ZackowskiKM and ThachWT. (2000). Cerebellar ataxia: torque deficiency or torque mismatch between joints?. J Neurophysiol, 83:3019–3030.
16.
KleinC, ButtSJ, MacholdRP, JohnsonJE and FishellG. (2005). Cerebellum- and forebrain-derived stem cells possess intrinsic regional character. Development, 132:4497–4508.
17.
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.
18.
WystrychowskiW, PatlollaB, ZhugeY, NeofytouE, RobbinsRC and BeyguiRE. (2016). Multipotency and cardiomyogenic potential of human adipose-derived stem cells from epicardium, pericardium, and omentum. Stem Cell Res Ther, 7:84.
19.
NakagomiT, TaguchiA, FujimoriY, SainoO, Nakano-DoiA, KuboS, GotohA, SomaT, YoshikawaH, et al. (2009). Isolation and characterization of neural stem/progenitor cells from post-stroke cerebral cortex in mice. Eur J Neurosci, 29:1842–1852.
20.
BhingeA, NambooriSC, ZhangX, VanDongenAMJ and StantonLW. (2017). Genetic correction of SOD1 mutant iPSCs reveals ERK and JNK activated AP1 as a driver of neurodegeneration in amyotrophic lateral sclerosis. Stem Cell Rep, 8:856–869.
21.
DraniasMR, JuH, RajaramE and VanDongenAM. (2013). Short-term memory in networks of dissociated cortical neurons. J Neurosci, 33:1940–1953.
22.
ReynoldsBA and WeissS. (1992). Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science, 255:1707–1710.
23.
NakataM, NakagomiT, MaedaM, Nakano-DoiA, MomotaY and MatsuyamaT. (2017). Induction of perivascular neural stem cells and possible contribution to neurogenesis following transient brain ischemia/reperfusion injury. Transl Stroke Res, 8:131–143.
24.
ShimadaIS, LeComteMD, GrangerJC, QuinlanNJ and SpeesJL. (2012). Self-renewal and differentiation of reactive astrocyte-derived neural stem/progenitor cells isolated from the cortical peri-infarct area after stroke. J Neurosci, 32:7926–7940.
25.
MokryJ, CizkovaD, FilipS, EhrmannJ, OsterreicherJ, KolarZ and EnglishD. (2004). Nestin expression by newly formed human blood vessels. Stem Cells Dev, 13:658–664.
26.
SuzukiS, NamikiJ, ShibataS, MastuzakiY and OkanoH. (2010). The neural stem/progenitor cell marker nestin is expressed in proliferative endothelial cells, but not in mature vasculature. J Histochem Cytochem, 58:721–730.
27.
Nakano-DoiA, NakagomiT, FujikawaM, NakagomiN, KuboS, LuS, YoshikawaH, SomaT, TaguchiA and MatsuyamaT. (2010). Bone marrow mononuclear cells promote proliferation of endogenous neural stem cells through vascular niches after cerebral infarction. Stem Cells, 28:1292–1302.
28.
NakagomiN, NakagomiT, KuboS, Nakano-DoiA, SainoO, TakataM, YoshikawaH, SternDM, MatsuyamaT and TaguchiA. (2009). Endothelial cells support survival, proliferation, and neuronal differentiation of transplanted adult ischemia-induced neural stem/progenitor cells after cerebral infarction. Stem Cells, 27:2185–2195.
29.
NewSE, Alvarez-GonzalezC, VagaskaB, GomezSG, BulstrodeNW, MadrigalA and FerrettiP. (2015). A matter of identity—phenotype and differentiation potential of human somatic stem cells. Stem Cell Res, 15:1–13.
30.
PehGS, LangRJ, PeraMF and HawesSM. (2009). CD133 expression by neural progenitors derived from human embryonic stem cells and its use for their prospective isolation. Stem Cells Dev, 18:269–282.
31.
TakagiT, YoshimuraS, SakumaR, Nakano-DoiA, MatsuyamaT and NakagomiT. (2017). Novel regenerative therapies based on regionally induced multipotent stem cells in post-stroke brains: their origin, characterization, and perspective. Transl Stroke Res, 8:515–528.
32.
EtcheversHC, VincentC, Le DouarinNM and CoulyGF. (2001). The cephalic neural crest provides pericytes and smooth muscle cells to all blood vessels of the face and forebrain. Development, 128:1059–1068.
33.
KornJ, ChristB and KurzH. (2002). Neuroectodermal origin of brain pericytes and vascular smooth muscle cells. J Comp Neurol, 442:78–88.
34.
LiP, DuF, YuellingLW, LinT, MuradimovaRE, TricaricoR, WangJ, EnikolopovG, BellacosaA, Wechsler-ReyaRJ and YangZJ. (2013). A population of nestin-expressing progenitors in the cerebellum exhibits increased tumorigenicity. Nat Neurosci, 16:1737–1744.
35.
WojcinskiA, LawtonAK, BayinNS, LaoZ, StephenDN and JoynerAL. (2017). Cerebellar granule cell replenishment postinjury by adaptive reprogramming of Nestin(+) progenitors. Nat Neurosci, 20:1361–1370.
36.
JacksonEL, Garcia-VerdugoJM, Gil-PerotinS, RoyM, Quinones-HinojosaA, VandenBergS and Alvarez-BuyllaA. (2006). PDGFR alpha-positive B cells are neural stem cells in the adult SVZ that form glioma-like growths in response to increased PDGF signaling. Neuron, 51:187–199.
37.
DoetschF, CailleI, LimDA, Garcia-VerdugoJM and Alvarez-BuyllaA. (1999). Subventricular zone astrocytes are neural stem cells in the adult mammalian brain. Cell, 97:703–716.
38.
KuhnHG, Dickinson-AnsonH and GageFH. (1996). Neurogenesis in the dentate gyrus of the adult rat: age-related decrease of neuronal progenitor proliferation. J Neurosci, 16:2027–2033.
39.
NakagomiT, MolnarZ, TaguchiA, Nakano-DoiA, LuS, KasaharaY, NakagomiN and MatsuyamaT. (2012). Leptomeningeal-derived doublecortin-expressing cells in poststroke brain. Stem Cells Dev, 21:2350–2354.
40.
NakagomiT, Nakano-DoiA and MatsuyamaT. (2015). Leptomeninges: a novel stem cell niche harboring ischemia-induced neural progenitors. Histol Histopathol, 30:391–399.
41.
DecimoI, BifariF, RodriguezFJ, MalpeliG, DolciS, LavariniV, PrettoS, VasquezS, SciancaleporeM, et al. (2011). Nestin- and doublecortin-positive cells reside in adult spinal cord meninges and participate in injury-induced parenchymal reaction. Stem Cells, 29:2062–2076.
42.
BifariF, BertonV, PinoA, KusaloM, MalpeliG, Di ChioM, BersanE, AmatoE, ScarpaA, et al. (2015). Meninges harbor cells expressing neural precursor markers during development and adulthood. Front Cell Neurosci, 9:383.
43.
BifariF, DecimoI, ChiamuleraC, BersanE, MalpeliG, JohanssonJ, LisiV, BonettiB, FumagalliG, PizzoloG and KramperaM. (2009). Novel stem/progenitor cells with neuronal differentiation potential reside in the leptomeningeal niche. J Cell Mol Med, 13:3195–3208.
44.
BifariF, DecimoI, PinoA, Llorens-BobadillaE, ZhaoS, LangeC, PanuccioG, BoeckxB, ThienpontB, et al. (2017). Neurogenic radial glia-like cells in meninges migrate and differentiate into functionally integrated neurons in the neonatal cortex. Cell Stem Cell, 20:360–373e367.
45.
NinomiyaS, EsumiS, OhtaK, FukudaT, ItoT, ImayoshiI, KageyamaR, IkedaT, ItoharaS and TamamakiN. (2013). Amygdala kindling induces nestin expression in the leptomeninges of the neocortex. Neurosci Res, 75:121–129.
46.
RoseSE, FrankowskiH, KnuppA, BerryBJ, MartinezR, DinhSQ, BrunerLT, WillisSL, CranePK, et al. (2018). Leptomeninges-derived induced pluripotent stem cells and directly converted neurons from autopsy cases with varying neuropathologic backgrounds. J Neuropathol Exp Neurol, 77:353–360.
47.
DolciS, PinoA, BertonV, GonzalezP, BragaA, FumagalliM, BonfantiE, MalpeliG, PariF, et al. (2017). High yield of adult oligodendrocyte lineage cells obtained from meningeal biopsy. Front Pharmacol, 8:703.
48.
BelmadaniA, RenD, BhattacharyyaBJ, RothwanglKB, HopeTJ, PerlmanH and MillerRJ. (2015). Identification of a sustained neurogenic zone at the dorsal surface of the adult mouse hippocampus and its regulation by the chemokine SDF-1. Hippocampus, 25:1224–1241.
49.
HutchingsM and WellerRO. (1986). Anatomical relationships of the pia mater to cerebral blood vessels in man. J Neurosurg, 65:316–325.
50.
KumarM, CsabaZ, PeineauS, SrivastavaR, RasikaS, ManiS, GressensP and El GhouzziV. (2014). Endogenous cerebellar neurogenesis in adult mice with progressive ataxia. Ann Clin Transl Neurol, 1:968–981.
KabaraM, KawabeJ, MatsukiM, HiraY, MinoshimaA, ShimamuraK, YamauchiA, AonumaT, NishimuraM, et al. (2014). Immortalized multipotent pericytes derived from the vasa vasorum in the injured vasculature. A cellular tool for studies of vascular remodeling and regeneration. Lab Invest, 94:1340–1354.
53.
BirbrairA, ZhangT, WangZM, MessiML, EnikolopovGN, MintzA and DelbonoO. (2012). Skeletal muscle pericyte subtypes differ in their differentiation potential. Stem Cell Res, 10:67–84.
54.
BirbrairA, ZhangT, WangZM, MessiML, EnikolopovGN, MintzA and DelbonoO. (2013). Role of pericytes in skeletal muscle regeneration and fat accumulation. Stem Cells Dev, 22:2298–2314.
55.
BirbrairA, ZhangT, WangZM, MessiML, MintzA and DelbonoO. (2014). Pericytes: multitasking cells in the regeneration of injured, diseased, and aged skeletal muscle. Front Aging Neurosci, 6:245.
56.
Farrington-RockC, CroftsNJ, DohertyMJ, AshtonBA, Griffin-JonesC and CanfieldAE. (2004). Chondrogenic and adipogenic potential of microvascular pericytes. Circulation, 110:2226–2232.
57.
DarA, DomevH, Ben-YosefO, TzukermanM, Zeevi-LevinN, NovakA, GermanguzI, AmitM and Itskovitz-EldorJ. (2012). Multipotent vasculogenic pericytes from human pluripotent stem cells promote recovery of murine ischemic limb. Circulation, 125:87–99.
58.
DohertyMJ, AshtonBA, WalshS, BeresfordJN, GrantME and CanfieldAE. (1998). Vascular pericytes express osteogenic potential in vitro and in vivo. J Bone Miner Res, 13:828–838.
59.
BirbrairA, ZhangT, WangZM, MessiML, OlsonJD, MintzA and DelbonoO. (2014). Type-2 pericytes participate in normal and tumoral angiogenesis. Am J Physiol Cell Physiol, 307:C25–C38.
60.
Nakano-DoiA, NakagomiT, SakumaR, TakahashiA, TanakaY, KawamuraM and MatsuyamaT. (2016). Expression patterns and phenotypic changes regarding stemness in brain pericytes in health and disease. J Stem Cell Res Ther, 6:332.
61.
KarowM, SanchezR, SchichorC, MasserdottiG, OrtegaF, HeinrichC, GasconS, KhanMA, LieDC, et al. (2012). Reprogramming of pericyte-derived cells of the adult human brain into induced neuronal cells. Cell Stem Cell, 11:471–476.
62.
KarowM, CampJG, FalkS, GerberT, PataskarA, Gac-SantelM, KageyamaJ, BrazovskajaA, GardingA, et al. (2018). Direct pericyte-to-neuron reprogramming via unfolding of a neural stem cell-like program. Nat Neurosci, 21:932–940.
63.
Nakano-DoiA, SakumaR, MatsuyamaT and NakagomiT. (2018). Ischemic stroke activates the VE-cadherin promoter and increases VE-cadherin expression in adult mice. Histol Histopathol, 33:507–521.
64.
PaulG, OzenI, ChristophersenNS, ReinbotheT, BengzonJ, VisseE, JanssonK, DannaeusK, Henriques-OliveiraC, et al. (2012). The adult human brain harbors multipotent perivascular mesenchymal stem cells. PLoS One, 7:e35577.
65.
CrisanM, YapS, CasteillaL, ChenCW, CorselliM, ParkTS, AndrioloG, SunB, ZhengB, et al. (2008). A perivascular origin for mesenchymal stem cells in multiple human organs. Cell Stem Cell, 3:301–313.
66.
BouacidaA, RossetP, TrichetV, GuillotonF, EspagnolleN, CordonierT, HeymannD, LayrolleP, SensebeL and DeschaseauxF. (2012). Pericyte-like progenitors show high immaturity and engraftment potential as compared with mesenchymal stem cells. PLoS One, 7:e48648.
67.
OzenI, BoixJ and PaulG. (2012). Perivascular mesenchymal stem cells in the adult human brain: a future target for neuroregeneration?. Clin Transl Med, 1:30.
68.
VezzaniB, PierantozziE and SorrentinoV. (2016). Not all pericytes are born equal: pericytes from human adult tissues present different differentiation properties. Stem Cells Dev, 25:1549–1558.
FengJ, MantessoA, De BariC, NishiyamaA and SharpePT. (2011). Dual origin of mesenchymal stem cells contributing to organ growth and repair. Proc Natl Acad Sci U S A, 108:6503–6508.
71.
CaplanAI. (2008). All MSCs are pericytes?. Cell Stem Cell, 3:229–230.
72.
CorselliM, ChenCW, CrisanM, LazzariL and PeaultB. (2010). Perivascular ancestors of adult multipotent stem cells. Arterioscler Thromb Vasc Biol, 30:1104–1109.
73.
UmeharaK, SunY, HiuraS, HamadaK, ItohM, KitamuraK, OshimaM, IwamaA, SaitoK, et al. (2018). A new conditionally immortalized human fetal brain pericyte cell line: establishment and functional characterization as a promising tool for human brain pericyte studies. Mol Neurobiol, 55:5993–6006.
74.
TakashimaY, EraT, NakaoK, KondoS, KasugaM, SmithAG and NishikawaS. (2007). Neuroepithelial cells supply an initial transient wave of MSC differentiation. Cell, 129:1377–1388.
75.
NagoshiN, ShibataS, NakamuraM, MatsuzakiY, ToyamaY and OkanoH. (2009). Neural crest-derived stem cells display a wide variety of characteristics. J Cell Biochem, 107:1046–1052.
76.
NagoshiN, ShibataS, KubotaY, NakamuraM, NagaiY, SatohE, MorikawaS, OkadaY, MabuchiY, et al. (2008). Ontogeny and multipotency of neural crest-derived stem cells in mouse bone marrow, dorsal root ganglia, and whisker pad. Cell Stem Cell, 2:392–403.
77.
KubotaY, TakuboK, HirashimaM, NagoshiN, KishiK, OkunoY, Nakamura-IshizuA, SanoK, MurakamiM, et al. (2011). Isolation and function of mouse tissue resident vascular precursors marked by myelin protein zero. J Exp Med, 208:949–960.
78.
CalloniGW, Le DouarinNM and DupinE. (2009). High frequency of cephalic neural crest cells shows coexistence of neurogenic, melanogenic, and osteogenic differentiation capacities. Proc Natl Acad Sci U S A, 106:8947–8952.
79.
CalloniGW, Glavieux-PardanaudC, Le DouarinNM and DupinE. (2007). Sonic hedgehog promotes the development of multipotent neural crest progenitors endowed with both mesenchymal and neural potentials. Proc Natl Acad Sci U S A, 104:19879–19884.
80.
Wislet-GendebienS, LaudetE, NeirinckxV, AlixP, LeprinceP, GlejzerA, PouletC, HennuyB, SommerL, ShakhovaO and RogisterB. (2012). Mesenchymal stem cells and neural crest stem cells from adult bone marrow: characterization of their surprising similarities and differences. Cell Mol Life Sci, 69:2593–2608.
81.
Simoes-CostaM and BronnerME. (2015). Establishing neural crest identity: a gene regulatory recipe. Development, 142:242–257.
82.
AlderJ, ChoNK and HattenME. (1996). Embryonic precursor cells from the rhombic lip are specified to a cerebellar granule neuron identity. Neuron, 17:389–399.
83.
MizuharaE, MinakiY, NakataniT, KumaiM, InoueT, MugurumaK, SasaiY and OnoY. (2010). Purkinje cells originate from cerebellar ventricular zone progenitors positive for Neph3 and E-cadherin. Dev Biol, 338:202–214.
84.
MossJ, GebaraE, BushongEA, Sanchez-PascualI, O'LaoiR, El M'GhariI, Kocher-BraissantJ, EllismanMH and ToniN. (2016). Fine processes of Nestin-GFP-positive radial glia-like stem cells in the adult dentate gyrus ensheathe local synapses and vasculature. Proc Natl Acad Sci U S A, 113:E2536–E2545.
85.
MakiT, MaedaM, UemuraM, LoEK, TerasakiY, LiangAC, ShindoA, ChoiYK, TaguchiA, et al. (2015). Potential interactions between pericytes and oligodendrocyte precursor cells in perivascular regions of cerebral white matter. Neurosci Lett, 597:164–169.
86.
SeoJH, MakiT, MaedaM, MiyamotoN, LiangAC, HayakawaK, PhamLD, SuwaF, TaguchiA, et al. (2014). Oligodendrocyte precursor cells support blood-brain barrier integrity via TGF-beta signaling. PLoS One, 9:e103174.
87.
ParkTI, MonzoH, MeeEW, BerginPS, TeohHH, MontgomeryJM, FaullRL, CurtisMA and DragunowM. (2012). Adult human brain neural progenitor cells (NPCs) and fibroblast-like cells have similar properties in vitro but only NPCs differentiate into neurons. PLoS One, 7:e37742.
88.
MakiharaN, ArimuraK, AgoT, TachibanaM, NishimuraA, NakamuraK, MatsuoR, WakisakaY, KurodaJ, et al. (2015). Involvement of platelet-derived growth factor receptor beta in fibrosis through extracellular matrix protein production after ischemic stroke. Exp Neurol, 264:127–134.
89.
VanlandewijckM, HeL, MaeMA, AndraeJ, AndoK, Del GaudioF, NaharK, LebouvierT, LavinaB, et al. (2018). A molecular atlas of cell types and zonation in the brain vasculature. Nature, 554:475–480.
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.
