Neuro-Muscular Differentiation of Adult Porcine Skin Derived Stem Cell-Like Cells 英文参考文献.docVIP

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Neuro-Muscular Differentiation of Adult Porcine Skin Derived Stem Cell-Like Cells 英文参考文献.doc

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Neuro-Muscular Differentiation of Adult Porcine Skin Derived Stem Cell-Like Cells 英文参考文献

Neuro-MuscularDifferentiationofAdultPorcineSkin DerivedStemCell-LikeCells DominikLermen1*,ErwinGorjup2,PaulW.Dyce3,HagenvonBriesen2,PaulMu¨ller1 1DepartmentofBiogeography,TrierUniversity,Trier,Germany,2DepartmentofCellBiologyandAppliedVirology,FraunhoferInstituteforBiomedicalEngineering,St. Ingbert,Germany,3DepartmentofAnimalandPoultryScience,UniversityofGuelph,Guelph,Ontario,Canada Abstract Background: Due to the genetic relationship to humans, porcine stem cells are a very important model system to investigatecelldifferentiation,associatedcellsignalingpathways,andcellfate.Porcineskinderivedstemcellshavebeen isolated from mid-gestation porcine fetus recently. To our knowledge, stem cells from the skin of the adult porcine organism have not been isolated until now. Hence, to our knowledge, we here describe the isolation, expansion, characterizationanddifferentiationofmultipotentporcineskinderivedstemcell-likecells(pSSCs)fromtheadultporcine organismforthefirsttime. Methodology/PrincipalFindings:pSSCshadaspindleshapedmorphologysimilartomesenchymalstemcells(MSCs).They couldbemaintainedproliferativelyactiveinvitroformorethan120daysandwereabletoformcoloniesfromsinglecells. pSSCsexpressedSox2andOct3/4,bothtranscriptionfactorsessentialtothepluripotentandself-renewingphenotypesof embryonicstemcells,whichrecentlygainedattentionduetotheirfunctionininducingpluripotentstemcells.Furthermore, theexpressionoftheprogenitormarkernestin,thesomaticstemcellmarkersBcrp1/ABCG2,Bmi1,andStat3wasdetected by reverse transcriptase-polymerase chain reaction (RT-PCR) in undifferentiated pSSCs. Flow cytometry revealed the expressionoftheMSCrelatedproteinsCD9,CD29,CD44andCD105,butnotCD90.Afterneuronaldifferentiationcellswith a characteristic morphology of neuronal and smooth muscle-like cells were present in the cultures. Subsequent immunochemistryandflowcytometryrevealedthedown-regulationofnestinandtheup-regulationoftheneuronspecific protein beta-III-tubulin and the astrocyte marker GFAP. Als