evidences

RegulationofNPCDifferentiationfromBoneMarrowStromal

CellsbyConstructionofNovelInducingSystem

Abstract:

Neuralprecursorcells(NPC)exploitedas“edcells”intissueengineering

aretheperfectcellsourceforthetreatmentofdegenerativediasandtraumaof

r,thecellsaresomewhatlimitedintheirutility

duetosource-relatedethicsorlegislations,andthediffcultiesinacquiringfrom

autologoustissue,suggestingdiscoveryanddevelopmentofnovelapproachthat

ntyears,theutilityofbone

marrowderivedmenchymalstemcells(BMSCs)todifferentiateintoneural

singrearch

trendtoutilizegrowthfactors(GFs)asaninducertoevokeBMSCsforneural

differentiation,howevertherelateddifferetiationmechanismsinducedbyGFsare

ly,manyevidenceshaveprovedthatPI3K/AKT

tion,it

hasalsobeendemonstratedthatGFsplayanimportantroleintheactivationof

PI3K/orewepropoahypothesisthat

mechanismsinvolvedinthedifferetiationofMSCsintoneuroninducedbyGFsis

meditatedbyitssubquentinductionofPI3K/n

themechanism,wecouldattempttoutilizeneuroblastomacells(NBCs)asa

potentialGFspoolfortheconstantcrectionofmanyGFs,whichmayjointly

activtethissignalingpathwayandefficientlypromotetheneuraldifferentiationof

therhand,bFGF,onetypeofGFs,isdemonstratedtobeableto

eruthe

NBCsandbFGF,wecouldutilizethetechniqueofmultilayeredalginate

microcapsulestomakethecoalitionofthemfortheNPCdifferentiationof

l,wehypothesisthatNBCsmicrocapsulatedinmultilayered

alginatemicrocapsuleswithbFGFsupplementcouldefficientlyinducethe

differentiationofMSCsintoNPCsthroughPI3K/AKTsignalingpathway

ible,thishypothesismightopenanattractive

approachforclinicalNPCtransfortationtoboostneuronregenerationandsynap

reconstruction,andprovidetherelevanttheoreticalbasisfortreatmentofcentral

nervoussystemdias.

Intruduction

Neuro-degenerativediasandcentralnervoussystemtraumahavebeen

ited

potentialsofcentralnervoussysteminnerveregenerationandfunction

reconstructioniscurrentlyahardnuttocrackforthetreatmentofthedias.

Transplantionofstemcells,particularlyneuralprecursorcells(NPCs)/neuralstem

cells(NSCs)havebeenpropodasapromisingtherapeuticstrategyforneuron

repairation[1].PlentyofstudiesidentifiesthatNPCscouldfacilitateCNS

regenerationduetotheirdifferentiationplasticity,whichmightleadtothe

substitutionofdeadcellsandfunctionalintegration[2-3].Itisalsoprovedthat

NPCsposssthecapacitytoreleaneuroprotectivefactorswhicharecapableof

stimulatingendogenousregeneration[4-5].However,thereliablesourceofNPCsis

ntyears,thedifferentiationof

BMSCsintoneuralcellshasbecomeafocusofcurrenttissueengineering

rearches[6-8].BMSCsthatarecapableoflf-renewal[9-10]havebeenwidely

demonstratedtoposssthecapacityofneuraldifferentiationintheprenceof

GFs[11-12].Inaddition,someliteraturesconfirmedthatMSCscanbeconverted

intoNSCs/NPCsinvitrounderthemulti-cellfactors[13-14].However,the

mechanismsinvolvedinthedifferetiationofMSCintoneuroninducedbyGFsis

therhand,althoughMSCshavetheabilitytodifferentiateinto

nervecells,thedifferentiationrateswererelativelylowinmostexperiments,

especially,orethispapercould

providesomecorrelativehypothesistodiscusstheissues.

ectofgrowthfactorsonMSCsdifferetiationisassociatedwith

activationofPI3k/AKTsignalingpathway

Recently,variousdatasuggestedthatveralintracellularsignaltransduction

pathwaysincludingPI3K/AKT,cAMP-PKAandJAK-STAT3signalingpathway

couldtriggerandcontroltheneuronaldifferentiationofMSCs[15-17].Particularly,

PI3K/AKTsignalinghasbeenrevealedtopossstheadditionactionofregulating

neuritedevelopmentandneuroprotection[18-20].Therefore,wefocudonthe

relationshipbetweenPI3K/AKTsignalingpathwayandneuronaldifferetiationof

MSC.

PI3K(phosphoinositol-3-kinas),afamilyofkinas,couldgeneratePIP2

intoPIP3(phosphoinositide-3,4,5-triphosphate),whichisacondmesngerthat

activatesmanytargetmoleculesincludingAKT/3K/AKTsignaling

pathwayhasbeenshowntocontributetotheregulationofBMSCsdifferentiation

throughup-regulationofproteinsthatmediatecellsurvivalsuchasmammalian

targetofrapamycin(mTOR)[21].Inaddition,itisreportedthattheactivityof

PI3K/AKTsignallingpathwaycouldbeevokedbyvariousGFssuchas

platelet-derivedgrowthfactor(PDGF),EpidermalGrowthFactor(EGF),basic

fibroblastgrowthfactor(bFGF),andVEGF[19-22].Thefindings,togetherwith

theobrvationsfromHermannetal[13]andHuWeietal[14]thattestifiedthe

indispensableroleofmulti-GFsinNPCs/,

wehypothesisthatthemechanismsinvolvedinthedifferentiationofMSCsinto

neuroninducedbyGFsismediatedbyitssubquentinductionofPI3K/AKT

themechanisms,maybewecouldutilizethesuperiority

ofimmortalcelllinessuchasNBCstocontinuouslycretemanyGFsin

Micro-environment，theallianceofGFsjointlyactivtethissignalingpathwayand

contributetothegrowthdifferentiationofBMSCsintoneuralcells.

ianceofGFscretedbyNBCsareinvovedinregulationofMSCs

differentiation

Neuroblastomaisakindofneuroendocrinetumor,whichderivedfromthe

eenshown

actorssuchasVEGF-B,

VEGF-C,bFGF,PDGFplayanimportantroleincellgrowth,progressionand

metastasisofNB[23-24],duetotheireffectofregulatingcellproliferationand

differentiation.

Theroleofconditionedmedium(CM)fromNBCsinthemodificationofcell

ve

demonstratedthatthedifferetiationofoligodendrocyteprecursorcellscouldbe

availablebyinducingneuralstemcellsintheprenceofconditionedmedium(CM)

obtainedfromB104neuroblastomacells(B104-CM),bFGFandPDGF-AAin

B104CMare2keyfactorsthatstimulateoligodendrocyteprecursorcell(OPC)

proliferationanddifferentiation[25].Moreover,veshownthat

conditionedmedium(CM)ofB104neuroblastomacellswascapableofinducing

differentiationofadipotissue-derivedmenchymalstemcells(AT-MSCs)toa

neuronalphenotype[26].Thus,abovestudiessomeextentconfirmedthat

thecombinedeffectsofmultipleGFscretedbyNBCscouldtriggerandpromote

theneuraldifferentiation.

sueengineeringtechnologyofcellsmicrocapsulation

Badonthepotentialofconditionedmedium(CM)fromNBCsto

induceneuraldifferentiation,wesuggestanapproachtoobtainsufficientand

constantGFsfromconditionedmedium(CM)

microencapsulationasapromisingstrategyhaveachievedimportantprogressfor

capsulationtechnology,asthenameimplies,isan

idesthe

uniquecultivationof3Denvironmentandpromotesthepassageofoxygen,

therapeuticproteins,nutrients,growthfactorsduetoitsthinmembraneand

relativelyhighratioofsurface-to-volume[27].Cellencapsulationcanalsoprotect

thecellfromthehost’simmunesystemandmechanicalstress[28].Inadditionthe

technologycouldachievecontinuousdeliverofGFsbytheencapsulatedcellsover

time[29-30].Indeed,thecharacteristicsofmicroencapsulatedtumorcells(TCs)

edthat

thecultureofhepatocarcinomacellsinalginate-poly-lysine-alginate(APA)

microcapsulesinvitrocouldproliferateTCswithhighviability,forming

multicellularspheroidjustasthecytoarchitectureofTCsinvivo[31].ShaolingWu

thattheimplantsofmicroencapsulatedratpheochromocytoma(PC12)

cellscouldcretecatecholaminespeciallydopaminetodecreathe

coldallodyniabehaviorofratsthathadtheneuropathicpain[32].Theyalsofound

thatthecell-loadedgroupdidnotoccurneoplasiaintherats’spinalcords[33].

Therefore,wehypothesisthatthesuperiorityofcellmicroencapsulationtechnique

cannotonlyenhanceNBCsproliferationandboosttheeffectontheneural

differentiaonofMSCs,butalsogregateNBCsfromnormalcells,thusreducing

theirharmfuleffectofcarcinogenicity.

gyofmicroencapsulatedNBCstogetherwithbFGFsupplementfor

NPC/NSCdifferetionderiveredfromMSCs(sig.1)

Severalexperimentshavefoundthattheexpressionofnestin(amarkerof

neuralstemcells)graduallydecreaintheprocessoftheneuronaldifferetiation

ofMSCs,indicatingthatNPC/NSCderiveredfromMSCdifferentiatedintomature

neuralcells(MNCs)F,onetypeofGFs,hasbeenreportedto

beabletoprolongthetransformationofNPCsintomatureneuralcellsandleadto

neuronalcellsdedifferentiateandappearsimilartoneuralstemcells[34],

suggestingtheutilizeofbFGFcouldstimulatethedifferetiationofNPC/NSC

ly,bFGFhasalsobeenudasthethemajor

nthedata,wehypothesisthe

specialcharacteristicofbFGFmaybeappliedtotheissueofNPC/NSC

er,accordingtothetechniqueofcontemporary

tissueengineering,wecanprojecteaproceduretosynthesizemultilayeredalginate

system,Thealginatecore

wascoatedwithapermlectivepoly-L-ornithine(PLO)layerwhichcould

facilitatethepermeabilityofsolutesintothemicrocapsules[35],theinnerlayer

canbeudforNBencapsulation,whiletheouterlayercanbeutilizedforbFGF

portedthattheadditionoftheouterlayercouldkeepthe

bFGFsustainedreleaformorethan18dayswithoutalteringthepermlectivity

ofthemicrocapsulecoat[36].MeanwhilethebFGFalsocouldpromotethe

capacityofproliferationofNBCs,togetherwiththeabilityofNBCstodeliverythe

bFGF,othemutualpromotion

betweenNBCsandbFGFintheinductionsystemmayplayaprominentpartin

NPC/NSCdifferetiationderiveredfromMSCs.

Conclusion:

Recently,thetissueengineeringtechnologyofneuraldifferetiationfrom

BMSCsappearsbrightdevelopingprospectsintreatmentforcentralnervous

rocessofneuronaldifferentiationofMSCs,PI3K/AKT

signalingpathwaythatcouldbeactivatedbyGFshavebeenprovedtoplayan

esntialroleinthistransformation,suggestingahypothesisthatGFscould

activateitssubquentinductionofPI3K/AKTsignalingpathwayandpromote

thismechanism,weexploitto

co-culturethemicroencapsulationofNBCandbFGFwithMSCsforpromotionof

elinductionsystemmaygreatlystimulatethe

directeddifferetiationrateofNPC/r,

subquentexperimentalstudiesarerequiredtofurthervalidatethepotential

benefitsofthenovelinductionsystem.

Conflictofinterest

None.

Figure1

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