Name: progenitor-derived oligodendrocyte culture system from human fetal brain
Uploaded: 2012-12-20T11:00:00
Description: Watch this Scientific Journal Video about Progenitor-derived Oligodendrocyte Culture System from Human Fetal Brain at JoVE.com

Denne forskning blev støttet af Intramural Research Program ved National Institutes of Health, NINDS. Forfatterne vil gerne takke alle medlemmerne af Laboratory of Molecular Medicine og Neurovidenskab, Rick Dreyfuss for at hjælpe med mikroskopi og Pamela C. Sigtning for at hjælpe med redigeringen.

Bemærk: Til rutinemæssig dyrkning af neurale stamceller og oligodendrocytic afstamningsceller, er inkubation udføres ved 37 ° C i en befugtet 5% CO2-atmosfære. Hver 2 dage, udskiftes mediet med 50 til 100% frisk medium, hvis kultur er 40-70% sammenflydende. På tidspunktet for nær sammenløb, er kulturerne passeres på 2-2.5e6/T75 kolbe som regel på et ugeskema. 1. Fremstilling af de overtrukne Flask <ol><li> Til fremstilling af overtrukne kolber fortynde...

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	<li>Messam, C. A., Hou, J., Gronostajski, R. M., Major, E. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lineage+pathway+of+human+brain+progenitor+cells+identified+by+JC+virus+susceptibility.">Lineage pathway of human brain progenitor cells identified by JC virus susceptibility.</a> Ann. Neurol. 53, 636-646 (2003).</li><li>Vescovi, A. L., Reynolds, B. A., Fraser, D. D., Weiss, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=bFGF+regulates+the+proliferative+fate+of+unipotent+(neuronal)+and+bipotent+(neuronal/astroglial)+EGF-generated+CNS+progenitor+cells.">bFGF regulates the proliferative fate of unipotent (neuronal) and bipotent (neuronal/astroglial) EGF-generated CNS progenitor cells.</a> Neuron. 11, 951-966 (1993).</li><li>Johe, K. K., Hazel, T. G., Muller, T., Dugich-Djordjevic, M. M., McKay, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+factors+direct+the+differentiation+of+stem+cells+from+the+fetal+and+adult+central+nervous+system.">Single factors direct the differentiation of stem cells from the fetal and adult central nervous system.</a> Genes Dev. 10, 3129-3140 (1996).</li><li>McKay, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stem+cells+in+the+central+nervous+system.">Stem cells in the central nervous system.</a> Science. 276, 66-71 (1997).</li><li>Belachew, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postnatal+NG2+proteoglycan-expressing+progenitor+cells+are+intrinsically+multipotent+and+generate+functional+neurons.">Postnatal NG2 proteoglycan-expressing progenitor cells are intrinsically multipotent and generate functional neurons.</a> J. Cell Biol. 161, 169-186 (2003).</li><li>Almazan, G., McKay, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+oligodendrocyte+precursor+cell+line+from+rat+optic+nerve.">An oligodendrocyte precursor cell line from rat optic nerve.</a> Brain Res. 579, 234-245 (1992).</li><li>Filipovic, R., Zecevic, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroprotective+role+of+minocycline+in+co-cultures+of+human+fetal+neurons+and+microglia.">Neuroprotective role of minocycline in co-cultures of human fetal neurons and microglia.</a> Exp. Neurol. 211, 41-51 (2008).</li><li>Goldman, S. A., Natesan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+niche-defying+feat:+induced+oligoneogenesis+in+the+adult+dentate+gyrus.">A niche-defying feat: induced oligoneogenesis in the adult dentate gyrus.</a> Cell Stem Cell. 3, 125-126 (2008).</li><li>Nunes, M. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+isolation+of+multipotential+neural+progenitor+cells+from+the+subcortical+white+matter+of+the+adult+human+brain.">Identification and isolation of multipotential neural progenitor cells from the subcortical white matter of the adult human brain.</a> Nat. Med. 9, 439-447 (2003).</li><li>Lyssiotis, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+histone+deacetylase+activity+induces+developmental+plasticity+in+oligodendrocyte+precursor+cells.">Inhibition of histone deacetylase activity induces developmental plasticity in oligodendrocyte precursor cells.</a> Proc. Natl. Acad. Sci. U.S.A. 104, 14982-14987 (2007).</li><li>Sim, F. J., Goldman, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=White+matter+progenitor+cells+reside+in+an+oligodendrogenic+niche.">White matter progenitor cells reside in an oligodendrogenic niche.</a> Ernst Schering Res. Found Workshop. , 61-81 (2005).</li><li>Raff, M. C., Miller, R. H., Noble, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+glial+progenitor+cell+that+develops+in+vitro+into+an+astrocyte+or+an+oligodendrocyte+depending+on+culture+medium.">A glial progenitor cell that develops in vitro into an astrocyte or an oligodendrocyte depending on culture medium.</a> Nature. 303, 390-396 (1983).</li><li>Windrem, M. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progenitor+cells+derived+from+the+adult+human+subcortical+white+matter+disperse+and+differentiate+as+oligodendrocytes+within+demyelinated+lesions+of+the+rat+brain.">Progenitor cells derived from the adult human subcortical white matter disperse and differentiate as oligodendrocytes within demyelinated lesions of the rat brain.</a> J. Neurosci. Res. 69, 966-975 (2002).</li><li>Chen, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+culture+of+rat+and+mouse+oligodendrocyte+precursor+cells.">Isolation and culture of rat and mouse oligodendrocyte precursor cells.</a> Nat. Protoc. 2, 1044-1051 (2007).</li><li>Armstrong, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+immature+oligodendrocyte+lineage+cells.">Isolation and characterization of immature oligodendrocyte lineage cells.</a> Methods. 16, 282-292 (1998).</li><li>Hu, B. Y., Du, Z. W., Li, X. J., Ayala, M., Zhang, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+oligodendrocytes+from+embryonic+stem+cells:+conserved+SHH+signaling+networks+and+divergent+FGF+effects.">Human oligodendrocytes from embryonic stem cells: conserved SHH signaling networks and divergent FGF effects.</a> Development. 136, 1443-1452 (2009).</li><li>Gard, A. L., Williams, W. C., Burrell, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oligodendroblasts+distinguished+from+O-2A+glial+progenitors+by+surface+phenotype+(O4+GalC-)+and+response+to+cytokines+using+signal+transducer+LIFR+beta.">Oligodendroblasts distinguished from O-2A glial progenitors by surface phenotype (O4+GalC-) and response to cytokines using signal transducer LIFR beta.</a> Dev. Biol. 167, 596-608 (1995).</li><li>Hu, B. Y., Du, Z. W., Zhang, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differentiation+of+human+oligodendrocytes+from+pluripotent+stem+cells.">Differentiation of human oligodendrocytes from pluripotent stem cells.</a> Nat. Protoc. 4, 1614-1622 (2009).</li><li>Reubinoff, B. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+progenitors+from+human+embryonic+stem+cells.">Neural progenitors from human embryonic stem cells.</a> Nat. Biotechnol. 19, 1134-1140 (2001).</li><li>Pfeiffer, S. E., Warrington, A. E., Bansal, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+oligodendrocyte+and+its+many+cellular+processes.">The oligodendrocyte and its many cellular processes.</a> Trends Cell Biol. 3, 191-197 (1993).</li><li>Zhang, S. C., Ge, B., Duncan, I. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tracing+human+oligodendroglial+development+in+vitro.">Tracing human oligodendroglial development in vitro.</a> J. Neurosci. Res. 59, 421-429 (2000).</li><li>Bradl, M., Lassmann, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oligodendrocytes:+biology+and+pathology.">Oligodendrocytes: biology and pathology.</a> Acta Neuropathol. 119, 37-53 (2010).</li><li>Chong, S. Y., Chan, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tapping+into+the+glial+reservoir:+cells+committed+to+remaining+uncommitted.">Tapping into the glial reservoir: cells committed to remaining uncommitted.</a> J. Cell Biol. 188, 305-312 (2010).</li><li>Jakovcevski, I., Filipovic, R., Mo, Z., Rakic, S., Zecevic, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oligodendrocyte+development+and+the+onset+of+myelination+in+the+human+fetal+brain.">Oligodendrocyte development and the onset of myelination in the human fetal brain.</a> Front Neuroanat. 3, 5 (2009).</li><li>Rao, R. C., Boyd, J., Padmanabhan, R., Chenoweth, J. G., McKay, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+serum-free+derivation+of+oligodendrocyte+precursors+from+neural+stem+cell-enriched+cultures.">Efficient serum-free derivation of oligodendrocyte precursors from neural stem cell-enriched cultures.</a> Stem Cells. 27, 116-125 (2009).</li><li>D'Intino, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Triiodothyronine+administration+ameliorates+the+demyelination/remyelination+ratio+in+a+non-human+primate+model+of+multiple+sclerosis+by+correcting+tissue+hypothyroidism.">Triiodothyronine administration ameliorates the demyelination/remyelination ratio in a non-human primate model of multiple sclerosis by correcting tissue hypothyroidism.</a> J. Neuroendocrinol. 23, 778-790 (2011).</li><li>Cui, Q. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+fetal+oligodendrocyte+progenitor+cells+from+different+gestational+stages+exhibit+substantially+different+potential+to+myelinate.">Human fetal oligodendrocyte progenitor cells from different gestational stages exhibit substantially different potential to myelinate.</a> Stem Cells Dev. , (2012).</li><li>Messam, C. A., Hou, J., Major, E. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coexpression+of+nestin+in+neural+and+glial+cells+in+the+developing+human+CNS+defined+by+a+human-specific+anti-nestin+antibody.">Coexpression of nestin in neural and glial cells in the developing human CNS defined by a human-specific anti-nestin antibody.</a> Exp. Neurol. 161, 585-596 (2000).</li></ol>

Denne protokol beskriver, hvordan man kan udlede føtale oligodendrocytter fra primære humane neurale stamceller og karakterisere deres fænotype ved hjælp af både flowcytometri og immunfluorescensfarvning. Den ekspansion og vækst af neurale stamceller fra fostre CNS har været meget godt beskrevet 1-4. Men opnåelse af tilstrækkelige menneskelige oligodendrocytter til in vitro-eksperimenter stadig vanskeligt, selv om det er muligt at identificere og isolere gliale precursorer fra vokse...

<table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalognummer </td><td> Slutkoncentration </td></tr><tr><td> DME / Hams F12 01:01 </td><td> Omega Scientist </td><td> DM-251 </td><td> 1X </td></tr><tr><td> Oksealbumin </td><td> Sigma </td><td> A9418 </td><td> 1% </td></tr><tr><td> Gentamicin </td><td> Kvalitet Biologicals </td><td> 120-098-031 </td><td> 50 ug / ml </td></tr><tr><td> L-Glutamin </td><td> Kvalitet Biologicals </td><td> 118-084-061 </td><td> 2 mM </td></tr><tr><td> T3 </td><td> Sigma </td><td> T2877 </td><td> 3 nM </td></tr><tr><td> N2 Components </td><td> Gibco BRL </td><td> 17.502 </td><td> 1:100 </td></tr><tr><td> NT-3 </td><td> PeproTech Inc </td><td> 450-03 </td><td> 2 ng / ml </td></tr><tr><td> Shh </td><td> R &amp; D System </td><td> 1314-SH/CF </td><td> 2 ng / ml </td></tr><tr><td> bFGF </td><td> PeproTech Inc </td><td> 100-18B </td><td> 20 ng / ml </td></tr><tr><td> PDGF-AA </td><td> PeproTech Inc </td><td> 100-13A </td><td> 10 ng / ml </td></tr><tr><td> PDL </td><td> Sigma </td><td> P6407 </td><td> 50 ug / m </td></tr><tr><td> PFA </td><td> Elektronmikroskopi Sciences </td><td> 15.712 </td><td> 2% </td></tr><tr><td> Trypsin </td><td> Kvalitet Biologicals </td><td> 118-087-721 </td><td></td></tr><tr><td> Papain </td><td> Worthington </td><td> LK003178 </td><td> 20 U / ml </td></tr><tr><td> DNase hætteglas </td><td> Worthington </td><td> LK003172 </td><td> 0,005% </td></tr><tr><td> EBSS </td><td> Worthington </td><td> LK003188 </td><td></td></tr><tr><td> Forlæng Guld medDAPI </td><td> Invitrogen </td><td> P36931 </td><td></td></tr><tr><td></td><td></td><td></td><td> Tabel 1. Reagenser. </td></tr><tr><td></td><td></td><td></td><td><table border="1"><tbody><tr><td> Primær Abs (alle ved 1 ug / ml) </td><td></td><td> Arter Isotype </td><td> Kilde </td><td> Sekundær Abs </td></tr><tr><td colspan="5"> Flowcytometri </td></tr><tr><td> A2B5-Biotin </td><td> Mus </td><td> IgM </td><td> Gift J. Nielson </td><td> Streptavidin PETR, Invitrogen, CA </td></tr><tr><td> O4FITC </td><td> Mus </td><td> IgM </td><td> Gave fra J. Nielson </td><td></td></tr><tr><td> A2B5 </td><td> Mus </td><td> IgM </td><td> Millipore, MA </td><td> GaM IgM-FITC, Invitrogen, CA </td></tr><tr><td> O4 </td><td> Mus </td><td> IgM </td><td> Millipore, MA </td><td> GaM IgM-FITC, Invitrogen, CA </td></tr><tr><td> GalC </td><td> Mus </td><td> IgG 3 </td><td> Millipore, MA </td><td> GaM IgG 3-PE, Southern Biotech, AL </td></tr><tr><td> MBP </td><td> Kylling </td><td> IgY </td><td> Millipore, MA </td><td> dαck IgY-AMCA, Jackson Immu., PA </td></tr><tr><td> Nestin </td><td> Mus </td><td> IgG1 </td><td> Messam et al. 2000 28 </td><td> GaM-IgG 1-PECy5, Invitrogen, CA </td></tr><tr><td> GFAP </td><td> Kanin </td><td> IgG </td><td> Millipore, MA </td><td> gαrb IgG-PE, Jackson Immu, PA </td></tr><tr><td> βIII tubulin </td><td> Mus </td><td> LgG2 </td><td> Covance, CA </td><td> GaM-IgG 2a-PETR, Invitrogen, CA </td></tr><tr><td colspan="6"> Immuncytokemi </td></tr><tr><td> βIII tubulin (1:1,500) </td><td> Mus </td><td> IgG2a </td><td> Covance, CA </td><td> GaM-IgG 2a-FITC, Invitrogen, CA (1:1,000) </td></tr><tr><td> GFAP (1:1,000) </td><td> Kanin </td><td> IgG </td><td> Millipore, MA </td><td> gαrb IgG-FITC, Jackson Immu, PA (1:500) </td></tr><tr><td> MBP (1:50) </td><td> Kylling </td><td> IgY </td><td> Millipore, MA </td><td> dαck IgY-FITC, Jackson Immu, PA (1:100) </td></tr><tr><td> O4 (1:100) </td><td> Mus </td><td> IgM </td><td> Millipore, MA </td><td> GaM IgM-AF546, Invitrogen, CA (1: 100) </td></tr><tr><td> GalC (1:10) </td><td> Kanin </td><td> IgG </td><td> Millipore, MA </td><td> GaM IgM-AF750, Invitrogen, CA (1:100) </td></tr></tbody></table> . Tabel 2 Antistoffer (Abs), der anvendes til flowcytometri og immuncytokemi assays antistofkonjugater: PE, phycoerythrin; PETR, phycoerythrin Texas Red, AMCA, amino-methyl-coumarin-acetat, CY, cyanin, FITC, fluoresceinisothiocyanat Ig:. Immunglobulin. . AF: Alexa Fluor, GaM: ged anti-mus, gαrb: gede-anti-kanin, dαck: æsel-anti-kylling. </td></tr></tbody></table>

progenitor-derived oligodendrocyte culture system from human fetal brain

Differentiering af humane neurale progenitorer til neuronale og gliale celletyper en model for at undersøge og sammenligne molekylære regulering af neural cellelinie udvikling. In vitro ekspansion af neurale progenitorer fra føtalt CNS-væv er blevet godt karakteriseret. Trods identifikation og isolering af gliale progenitorer fra voksent menneske subcorticale hvide substans og udvikling af forskellige dyrkningsbetingelser til direkte differentiering af føtale neurale stamceller i myelin producerende oligodendrocytter, erhverve tilstrækkelige menneskelige oligodendrocytter til in vitro eksperimenter stadig vanskelig. Differentiering af galactocerebrosid + (GalC) og O4 + oligodendrocyt precursor eller progenitorceller (OPC) fra neurale precursorceller er blevet rapporteret ved hjælp af andet trimester føtal hjerne. Men disse celler ikke proliferere i fraværet af bæreceller herunder astrocytter og neuroner, og går tabt hurtigt over tid i kultur. Der er stadig behov for et dyrkningssystem til fremstilling af celler af oligodendrocyt-afstamning egnet til in vitro forsøg. Kultur af primære humane oligodendrocytter kan for eksempel være en nyttig model til at studere patogenesen af neurotrofiske infektiøse agenser såsom det menneskelige polyomavirus, JCV, at in vivo inficerer disse celler. Disse dyrkede celler kan også modeller for andre demyelinerende sygdomme i centralnervesystemet (CNS). Primær, human føtal hjerne-afledt, multipotentielle neurale stamceller formere sig in vitro og samtidig bevare evnen til at differentiere til neuroner (stamfader-afledte neuroner, PDN) og astrocytter (stamfader-afledte astrocytter, PDA) Denne undersøgelse viser, at neurale stamceller kan induceres at differentiere gennem mange af de faser af oligodendrocytic afstamning udvikling (stamfader-afledte oligodendrocytter, BOB). Vi kultur neurale progenitorceller i DMEM-F12 serumfrie medier supsuppleret med basisk fibroblastvækstfaktor (bFGF), blodpladeafledt vækstfaktor (PDGF-AA), Sonic hedgehog (Shh), neurotrofisk faktor 3 (NT-3), N-2 og triiodothyronin (T3). De dyrkede celler er passage ved 2.5e6 celler pr 75cm kolber hver ca syv dage. Under anvendelse af disse betingelser, opretholde hovedparten af ​​celler i kultur en morfologi karakteriseret ved nogle processer og udtrykker markører for præ-oligodendrocytceller såsom A2B5 og O-4. Når vi fjerne de fire vækstfaktorer (GF) (bFGF, PDGF-AA, Shh, NT-3), og der tilsættes konditionerede medier fra PDN, cellerne begynder at erhverve flere processer og udtrykker markører specifikke for oligodendrocyt-differentiering, såsom GalC og myelin basisk protein (MBP). Vi udførte fænotypisk karakterisering ved hjælp af flerfarvet flowcytometri til at identificere entydige markører for oligodendrocyt.

Det er meget vigtigt at starte differentieringsprocessen fra en 70% -80% konfluente neural progenitorcelle kultur (figur 1A). Mange celler vil dø ud efter erstatning af dyrkningsmediet fra progenitoren til oligo medium eftersom den omfatter specifikke vækstfaktorer. Dette indikerer, at væksten af neurale progenitorceller ikke er disponeret for en oligodendrocytic fænotype ikke vil blive støttet af det nye medie (figur 1B). Inkubering i oligo medium + GF i en uge resulterede i en me...

Watch this Scientific Journal Video about Progenitor-derived Oligodendrocyte Culture System from Human Fetal Brain at JoVE.com

Progenitor-derived Oligodendrocyte Culture System from Human Fetal Brain

Primær, humane føtale hjerne-afledte, multipotentielle progenitorceller formere sig<em&gt; In vitro</em&gt; Samtidig med at evnen til at differentiere til neuroner og astrocytter. Dette arbejde viser, at neurale stamceller kan induceres til at differentiere gennem stadier af oligodendrocytic linie ved konditionering med udvalgte vækstfaktorer.

Stamfader-afledt oligodendrocyt Culture System fra human føtal hjerne

Differentiation of human neural progenitors into neuronal and glial cell types offers a model to study and compare molecular regulation of neural cell ...

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Understanding blood-brain barrier function under physiological and pathophysiological conditions is critical for the development of new therapeutic ...

Migration, Chemo-Attraction, and Co-Culture Assays for Human Stem Cell-Derived Endothelial Cells and GABAergic Neurons

Migration, Chemo-Attraction, og Co-Kultur Assays for Human Stem Cell-Afledt endotelceller og GABAergic Neurons

Role of brain vasculature in nervous system development and etiology of brain disorders is increasingly gaining attention. Our recent studies have ...

Measuring and Manipulating Functionally Specific Neural Pathways in the Human Motor System with Transcranial Magnetic Stimulation

Måling og manipulation funktionelt specifikke neurale veje i det menneskelige motorsystem med tværkraniel magnetisk stimulation

Understanding interactions between brain areas is important for the study of goal-directed behavior. Functional neuroimaging of brain connectivity has ...

Generation of Oligodendrocytes and Oligodendrocyte-Conditioned Medium for Co-Culture Experiments

Generation af Oligodendrocytes og Oligodendrocyte-Conditioned Medium for co-kultur eksperimenter

In the central nervous system, oligodendrocytes are well-known for their role in axon myelination, that accelerates the propagation of action potentials ...