Fundamental cures of central nervous system (CNS) diseases are rarely achieved due to the low regenerative ability of the CNS. Recently, cell-based therapy using mesenchymal stem cells (MSCs) has been explored as an effective treatment for CNS diseases. Among the various tissue-derived MSCs, we have isolated human cranial bone-derived MSCs (cMSCs) in our laboratory. In addition, we have focused on simulated microgravity (MG) as a valuable culture environment of MSCs. However, detailed mechanisms underlying functional recovery from transplantation of MSCs cultured under MG conditions remain unclear. In this study, we investigated the therapeutic mechanisms of transplantation of cMSCs cultured under MG conditions in traumatic brain injury (TBI) model mice. Human cMSCs were cultured under 1G and MG conditions, and cMSCs cultured under MG conditions expressed significantly higher messenger RNA (mRNA) levels of hepatocyte growth factor (HGF) and transforming growth factor beta (TGF-β). In TBI model mice, the transplantation of cMSCs cultured under MG conditions (group MG) showed greater motor functional improvement compared with only phosphate-buffered saline administration (group PBS). Moreover, the protein expression levels of tumor necrosis factor alpha (TNF-α) and the Bcl-2-associated X protein (Bax)/b cell leukemia/lymphoma 2 protein (Bcl-2) ratio were significantly lower at brain injury sites in mice of group MG than those of group PBS. In addition, an in vitro study showed that the conditioned medium of cMSCs cultured under MG conditions significantly suppressed the cell death of NG108-15 cells exposed to oxidative or inflammatory stress through anti-inflammatory and antiapoptosis effects. These findings demonstrate that culturing cMSCs under simulated MG increases the neuroprotective effects, suggesting that simulated MG cultures may be a useful method for cell-based therapy strategies for CNS diseases.
Get full access to this article
View all access options for this article.
References
1.
CookD and GeneverP. (2013). Regulation of mesenchymal stem cell differentiation. Adv Exp Med Biol, 786:213–229.
2.
ChenY, TengFY and TangBL. (2006). Coaxing bone marrow stromal mesenchymal stem cells towards neuronal differentiation: progress and uncertainties. Cell Mol Life Sci, 63:1649–1657.
3.
CaplanAI and DennisJE. (2006). Mesenchymal stem cells as trophic mediators. J Cell Biochem, 98:1076–1084.
4.
WilkinsA, KempK, GintyM, HaresK, MallamE and ScoldingN. (2009). Human bone marrow-derived mesenchymal stem cells secrete brain-derived neurotrophic factor which promotes neuronal survival in vitro. Stem Cell Res, 3:63–70.
5.
DengW, ObrockaM, FischerI and ProckopDJ. (2001). In vitro differentiation of human marrow stromal cells into early progenitors of neural cells by conditions that increase intracellular cyclic AMP. Biochem Biophys Res Commun, 282:148–152.
6.
KomatsuK, HonmouO, SuzukiJ, HoukinK, HamadaH and KocsisJD. (2010). Therapeutic time window of mesenchymal stem cells derived from bone marrow after cerebral ischemia. Brain Res, 1334:84–92.
7.
ChoSR, KimYR, KangHS, YimSH, ParkCI, MinYH, LeeBH, ShinJC and LimJB. (2009). Functional recovery after the transplantation of neurally differentiated mesenchymal stem cells derived from bone marrow in a rat model of spinal cord injury. Cell Transplant, 18:1359–1368.
8.
HonmouO, HoukinK, MatsunagaT, NiitsuY, IshiaiS, OnoderaR, WaxmanSG and KocsisJD. (2011). Intravenous administration of auto serum-expanded autologous mesenchymal stem cells in stroke. Brain, 134:1790–1807.
9.
TseWT, PendletonJD, BeyerWM, EgalkaMC and GuinanEC. (2003). Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplantation, 75:389–397.
10.
JinHJ, BaeYK, KimM, KwonSJ, JeonHB, ChoiSJ, KimSW, YangYS, OhW, ChangJW. (2013). Comparative analysis of human mesenchymal stem cells from bone marrow, adipose tissue, and umbilical cord blood as sources of cell therapy. Int J Mol Sci, 14:17986–18001.
11.
De BariC, Dell'AccioF, TylzanowskiP and LuytenFP. (2001). Multipotent mesenchymal stem cells from adult human synovial membrane. Arthritis Rheum, 44:1928–1942.
12.
ShinagawaK, MitsuharaT, OkazakiT, TakedaM, YamaguchiS, MagakiT, OkuraY, UwatokoH, KawaharaY, YugeL and KurisuK. (2015). The characteristics of human cranial bone marrow mesenchymal stem cells. Neurosci Lett, 606:161–166.
13.
MeadB, LoganA, BerryM, LeadbeaterW and SchevenBA. (2014). Paracrine-mediated neuroprotection and neuritogenesis of axotomised retinal ganglion cells by human dental pulp stem cells: comparison with human bone marrow and adipose-derived mesenchymal stem cells. PLoS One, 9:e109305.
14.
PlettPA, AbonourR, FrankovitzSM and OrschellCM. (2004). Impact of modeled microgravity on migration, differentiation, and cell cycle control of primitive human hematopoietic progenitor cells. Exp Hematol, 32:773–781.
15.
YugeL, HideI, KumagaiT, KumeiY, TakedaS, KannoM, SugiyamaM and KataokaK. (2003). Cell differentiation and p38(MAPK) cascade are inhibited in human osteoblasts cultured in a three-dimensional clinostat. In Vitro Cell Dev Biol Anim, 39:89–97.
16.
YugeL, KajiumeT, TaharaH, KawaharaY, UmedaC, YoshimotoR, WuSL, YamaokaK, AsashimaM, KataokaK and IdeT. (2006). Microgravity potentiates stem cell proliferation while sustaining the capability of differentiation. Stem Cells Dev, 15:921–929.
17.
MitsuharaT, TakedaM, YamaguchiS, ManabeT, MatsumotoM, KawaharaY, YugeL and KurisuK. (2013). Simulated microgravity facilitates cell migration and neuroprotection after bone marrow stromal cell transplantation in spinal cord injury. Stem Cell Res Ther, 4:35.
18.
ImuraT, TomiyasuM, OtsuruN, NakagawaK, OtsukaT, TakahashiS, TakedaM, ShresthaL and KawaharaY. (2017). Hypoxic preconditioning increases the neuroprotective effects of mesenchymal stem cells in a rat model of spinal cord injury. Stem Cell Res Ther, 7:375.
19.
NakamuraT, NishizawaT, HagiyaM, SekiT, ShimonishiM, SugimuraA, TashiroK and ShimizuS. (1989). Molecular cloning and expression of human hepatocyte growth factor. Nature, 342:440–443.
20.
SunW, FunakoshiH and NakamuraT. (2002). Localization and functional role of hepatocyte growth factor (HGF) and its receptor c-met in the rat developing cerebral cortex. Brain Res Mol Brain Res, 103:36–48.
21.
ShangJ, DeguchiK, OhtaY, LiuN, ZhangX, TianF, YamashitaT, IkedaY, MatsuuraT, et al. (2011). Strong neurogenesis, angiogenesis, synaptogenesis, and antifibrosis of hepatocyte growth factor in rats brain after transient middle cerebral artery occlusion. J Neurosci Res, 89:86–95.
22.
WeidnerKM, BehrensJ, VandekerckhoveJ and BirchmeierW. (1990). Scatter factor: molecular characteristics and effect on the invasiveness of epithelial cells. J Cell Biol, 111:2097–2108.
23.
CatonA, HackerA, NaeemA, LivetJ, MainaF, BladtF, KleinR, BirchmeierC and GuthrieS. (2000). The branchial arches and HGF are growth-promoting and chemoattractant for cranial motor axons. Development, 127:1751–1766.
24.
GutierrezH, DolcetX, TolcosM and DaviesA. (2004). HGF regulates the development of cortical pyramidal dendrites. Development, 131:3717–3726.
25.
HossainMA, RussellJC, GomezR and LaterraJ. (2002). Neuroprotection by scatter factor/hepatocyte growth factor and FGF-1 in cerebellar granule neurons is phosphatidylinositol 3-kinase/Akt-dependent and MAPK/CREB-independent. J Neurochem, 81:365–378.
26.
HeF, WuLX, ShuKX, LiuFY, YangLJ, ZhouX, ZhangY, HuangBS, HuangD and DengXL. (2008). HGF protects cultured cortical neurons against hypoxia/reoxygenation induced cell injury via ERK1/2 and PI-3K/Akt pathways. Colloids Surf B Biointerfaces, 61:290–297.
27.
MosesHL, BranumEL, ProperJA and RobinsonRA. (1981). Transforming growth factor production by chemically transformed cells. Cancer Res, 41:2842–2848.
28.
JianH, ShenX, LiuI, SemenovM, HeX and WangXF. (2006). Smad3-dependent nuclear translocation of beta-catenin is required for TGF-beta1-induced proliferation of bone marrow-derived adult human mesenchymal stem cells. Genes Dev, 20:666–674.
29.
WatabeT and MiyazonoK. (2009). Roles of TGF-beta family signaling in stem cell renewal and differentiation. Cell Res, 19:103–115.
30.
YangEY and MosesHL. (1990). Transforming growth factor beta 1-induced changes in cell migration, proliferation, and angiogenesis in the chicken chorioallantoic membrane. J Cell Biol, 111:731–741.
31.
BeckLS, DeguzmanL, LeeWP, XuY, McFatridgeLA and AmentoEP. (1991). TGF-beta 1 accelerates wound healing: reversal of steroid-impaired healing in rats and rabbits. Growth Factors, 5:295–304.
EcheverryS, ShiXQ, HawA, LiuH, ZhangZW and ZhangJ. (2009). Transforming growth factor-beta1 impairs neuropathic pain through pleiotropic effects. Mol Pain, 5:16.
34.
HuangC, AkaishiS and OgawaR. (2012). Mechanosignaling pathways in cutaneous scarring. Arch Dermatol Res, 304:589–597.
35.
KannoT, TakahashiT, TsujisawaT, AriyoshiW and NishiharaT. (2007). Mechanical stress-mediated Runx2 activation is dependent on Ras/ERK1/2 MAPK signaling in osteoblasts. J Cell Biochem, 101:1266–1277.
36.
JiangJG and ZarnegarR. (1997). A novel transcriptional regulatory region within the core promoter of the hepatocyte growth factor gene is responsible for its inducibility by cytokines via the C/EBP family of transcription factors. Mol Cell Biol, 17:5758–5770.
37.
LiH, GadeP, XiaoW and KalvakolanuDV. (2007). The interferon signaling network and transcription factor C/EBP-beta. Cell Mol Immunol, 4:407–418.
38.
Ruiz-OrtegaM, Rodríguez-VitaJ, Sanchez-LopezE, CarvajalG and EgidoJ. (2007). TGF-beta signaling in vascular fibrosis. Cardiovasc Res, 74:196–206.
LucasSM, RothwellNJ and GibsonRM. (2006). The role of inflammation in CNS injury and disease. Br J Pharmacol, 147(Suppl 1):S232–S240.
41.
BayirH, KaganVE, BorisenkoGG, TyurinaYY, JaneskoKL, VagniVA, BilliarTR, WilliamsDL and KochanekPM. (2005). Enhanced oxidative stress in iNOS-deficient mice after traumatic brain injury: support for a neuroprotective role of iNOS. J Cereb Blood Flow Metab, 25:673–684.
42.
WielochT and NikolichK. (2006). Mechanisms of neural plasticity following brain injury. Curr Opin Neurobiol, 16:258–264.
43.
Basic KesV, SimundicAM, NikolacN, TopicE and DemarinV. (2008). Pro-inflammatory and anti-inflammatory cytokines in acute ischemic stroke and their relation to early neurological deficit and stroke outcome. Clin Biochem, 41:1330–1334.
44.
ChodobskiA, ZinkBJ and Szmydynger-ChodobskaJ. (2011). Blood-brain barrier pathophysiology in traumatic brain injury. Transl Stroke Res, 2:492–516.
45.
StahelPF, ShohamiE, YounisFM, KariyaK, OttoVI, LenzlingerPM, GrosjeanMB, EugsterHP, TrentzO, KossmannT and Morganti-KossmannMC. (2000). Experimental closed head injury: analysis of neurological outcome, blood-brain barrier dysfunction, intracranial neutrophil infiltration, and neuronal cell death in mice deficient in genes for pro-inflammatory cytokines. J Cereb Blood Flow Metab, 20:369–380.
46.
YenariMA, XuL, TangXN, QiaoY and GiffardRG. (2006). Microglia potentiate damage to blood-brain barrier constituents: improvement by minocycline in vivo and in vitro. Stroke, 37:1087–1093.
47.
FloydCL and LyethBG. (2007). Astroglia: important mediators of traumatic brain injury. Prog Brain Res, 161:61–79.
48.
WangPP, XieDY, LiangXJ, PengL, ZhangGL, YeYN, XieC and GaoZL. (2012). HGF and direct mesenchymal stem cells contact synergize to inhibit hepatic stellate cells activation through TLR4/NF-kB pathway. PLoS One, 7:e43408.
49.
BendinelliP, MatteucciE, DogliottiG, CorsiMM, BanfiG, MaroniP and DesiderioMA. (2010). Molecular basis of anti-inflammatory action of platelet-rich plasma on human chondrocytes: mechanisms of NF-κB inhibition via HGF. J Cell Physiol, 225:757–766.
50.
BenvenisteEN, TangLP and LawRM. (1995). Differential regulation of astrocyte TNF-alpha expression by the cytokines TGF-beta, IL-6 and IL-10. Int J Dev Neurosci, 13:341–349.
51.
ChenNF, HuangSY, ChenWF, ChenCH, LuCH, ChenCL, YangSN, WangHM and WenZH. (2013). TGF-β1 attenuates spinal neuroinflammation and the excitatory amino acid system in rats with neuropathic pain. J Pain, 14:1671–1685.
NeirinckxV, AgirmanG, CosteC, MarquetA, DionV, RogisterB, FranzenR and WisletS. (2015). Adult bone marrow mesenchymal and neural crest stem cells are chemoattractive and accelerate motor recovery in a mouse model of spinal cord injury. Stem Cell Res Ther, 6:211.
54.
PomièsP, BlaquièreM, MauryJ, MercierJ, GouziF and HayotM. (2016). Involvement of the FoxO1/MuRF1/Atrogin-1 signaling pathway in the oxidative stress-induced atrophy of cultured chronic obstructive pulmonary disease myotubes. PLoS One, 11:e0160092.
55.
DumitruCD, CeciJD, TsatsanisC, KontoyiannisD, StamatakisK, LinJH, PatriotisC, JenkinsNA, CopelandNG, KolliasG and TsichlisPN. (2000). TNF-alpha induction by LPS is regulated posttranscriptionally via a Tpl2/ERK-dependent pathway. Cell, 103:1071–1083.
HuZX, GengJM, LiangDM, LuoM and LiML. (2010). Hepatocyte growth factor protects human embryonic stem cell derived-neural progenitors from hydrogen peroxide-induced apoptosis. Eur J Pharmacol, 645:23–31.
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.
