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

Dit onderzoek werd ondersteund door de Intramurale Research Program van de National Institutes of Health, NINDS. De auteurs willen graag alle leden van het Laboratorium voor Moleculaire Geneeskunde en Neuroscience, Rick Dreyfuss bedanken voor het helpen met microscopie en Pamela C. Zeven voor het helpen met de redactie.

Opmerking: routine kweken van neurale voorlopercellen en oligodendrocytic lineage cellen incubatie wordt uitgevoerd bij 37 ° C in een bevochtigde 5% CO2 atmosfeer. Elke 2 dagen wordt het medium vervangen met 50 tot 100% vers medium als cultuur 40-70% confluent. Op het moment van bijna confluentie worden de kweken doorgekweekt op 2-2.5e6/T75 kolf meestal wekelijks schema. 1. Voorbereiden van de Coated Flask <ol><li> Te bereiden beklede flessen 5 mg van poly-D-ly...

<ol>
	<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>

Dit protocol wordt beschreven hoe u foetale oligodendrocyten ontlenen primaire menselijke neurale stamcellen en karakteriseren hun fenotype met behulp van zowel flowcytometrie en immunofluorescentie kleuring. De uitbreiding en groei van neurale voorlopercellen uit foetale centrale zenuwstelsel is zeer goed beschreven 1-4. Echter, om voldoende menselijke oligodendrocyten in vitro experimenten blijft moeilijk, hoewel het mogelijk om voorlopers en gliale isoleren van volwassen menselijke witte stof <sup...

<table border="1"><tbody><tr><td> Naam van het reagens </td><td> Vennootschap </td><td> Catalogusnummer </td><td> Eindconcentratie </td></tr><tr><td> DME / HAMS F12 01:01 </td><td> Omega Scientist </td><td> DM-251 </td><td> 1X </td></tr><tr><td> Runderalbumine </td><td> Sigma </td><td> A9418 </td><td> 1% </td></tr><tr><td> Gentamicine </td><td> Kwaliteit Biologicals </td><td> 120-098-031 </td><td> 50 ug / ml </td></tr><tr><td> L-Glutamine </td><td> Kwaliteit 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 Componenten </td><td> Gibco BRL </td><td> 17502 </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> O &amp; O-systeem </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> Electron Microscopy Sciences </td><td> 15712 </td><td> 2% </td></tr><tr><td> Trypsine </td><td> Kwaliteit Biologicals </td><td> 118-087-721 </td><td></td></tr><tr><td> Papaïne </td><td> Worthington </td><td> LK003178 </td><td> 20 U / ml </td></tr><tr><td> DNase flacons </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> ProLong Gold metDAPI </td><td> Invitrogen </td><td> P36931 </td><td></td></tr><tr><td></td><td></td><td></td><td> Tabel 1. Reagentia. </td></tr><tr><td></td><td></td><td></td><td><table border="1"><tbody><tr><td> Primaire Abs (alle bij 1 ug / ml) </td><td></td><td> Soorten Isotype </td><td> Bron </td><td> Secundaire Abs </td></tr><tr><td colspan="5"> Flow Cytometry </td></tr><tr><td> A2B5-Biotin </td><td> Muis </td><td> IgM </td><td> Gift J. Nielson </td><td> Streptavidine PETR, Invitrogen, CA </td></tr><tr><td> O4FITC </td><td> Muis </td><td> IgM </td><td> Geschenk van J. Nielson </td><td></td></tr><tr><td> A2B5 </td><td> Muis </td><td> IgM </td><td> Millipore, MA </td><td> gαm IgM-FITC, Invitrogen, CA </td></tr><tr><td> O4 </td><td> Muis </td><td> IgM </td><td> Millipore, MA </td><td> gαm IgM-FITC, Invitrogen, CA </td></tr><tr><td> GalC </td><td> Muis </td><td> IgG3 </td><td> Millipore, MA </td><td> gαm IgG 3-PE, Zuid-Biotech, AL </td></tr><tr><td> MBP </td><td> Kip </td><td> IgY </td><td> Millipore, MA </td><td> dαck IgY-AMCA, Jackson Immu., PA </td></tr><tr><td> Nestin </td><td> Muis </td><td> IgG1 </td><td> Messam et al.. 2000 28 </td><td> gαm IgG 1-PECy5, Invitrogen, CA </td></tr><tr><td> GFAP </td><td> Konijn </td><td> IgG </td><td> Millipore, MA </td><td> gαrb IgG-PE, Jackson Immu, PA </td></tr><tr><td> βIII tubuline </td><td> Muis </td><td> IgG2 </td><td> Covance, CA </td><td> gαm IgG 2 a-PETR, Invitrogen, CA </td></tr><tr><td colspan="6"> Immunocytochemie </td></tr><tr><td> βIII tubuline (1:1,500) </td><td> Muis </td><td> IgG2a </td><td> Covance, CA </td><td> gαm IgG2a-FITC, Invitrogen, CA (1:1,000) </td></tr><tr><td> GFAP (1:1,000) </td><td> Konijn </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> Kip </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> Muis </td><td> IgM </td><td> Millipore, MA </td><td> gαm IgM-AF546, Invitrogen, CA (1: 100) </td></tr><tr><td> GalC (1:10) </td><td> Konijn </td><td> IgG </td><td> Millipore, MA </td><td> gαm IgM-AF750, Invitrogen, CA (1:100) </td></tr></tbody></table> . Tabel 2 Antilichamen (Abs) voor flowcytometrie en immunocytochemie assays antilichaamconjugaten: PE, fycoerythrine, PETR, fycoerythrine Texas Red, AMCA, amino-methyl-cumarine-acetaat; Cy, cyanine, FITC, fluoresceïne-isothiocyanaat Ig:. Immunoglobuline. . AF: Alexa Fluor; gαm: geit anti-muis; gαrb: geit anti-konijn; dαck: ezel anti-kip. </td></tr></tbody></table>

progenitor-derived oligodendrocyte culture system from human fetal brain

Differentiatie van humane neurale stamcellen in neuronale en gliale celtypen een model te bestuderen en vergelijken moleculaire regulatie van neurale cellijn ontwikkeling. In vitro expansie van neurale voorlopercellen uit foetale CZS-weefsel is goed gekarakteriseerd. Ondanks de identificatie en isolatie van gliale voorlopercellen uit volwassen menselijk subcorticale witte stof en ontwikkeling van verschillende kweekomstandigheden direct differentiatie van neurale stamcellen in myeline producerende oligodendrocyten, het verkrijgen van voldoende menselijke oligodendrocyten in vitro experimenten blijft moeilijk. Differentiatie van galactocerebroside + (GalC) en O4 + oligodendrocyt precursor of progenitorcellen (OPC) van neurale precursoren is gemeld met behulp van het tweede trimester foetale hersenen. Echter, deze cellen niet prolifereren in afwezigheid van steuncellen zoals astrocyten en neuronen en worden snel verloren tijd in cultuur. De noodzaak de een kweeksysteem te produceren cellen van de oligodendrocyten lineage geschikt voor in vitro experimenten. Kweek van primaire humane oligodendrocyten kan bijvoorbeeld een nuttig model voor de pathogenese van neurotrope ziekteverwekkers zoals het menselijk polyomavirus, JCV dat in vivo infecteert deze cellen te bestuderen. Deze gekweekte cellen kunnen ook modellen van andere demyeliniserende ziekten van het centrale zenuwstelsel (CNS). Primaire, foetale hersenen afgeleide, multipotentiële neurale progenitorcellen prolifereren in vitro terwijl de kunnen differentiëren tot neuronen (progenitor-afgeleide neuronen, PDN) en astrocyten (progenitor-afgeleide astrocyten, PDA) Dit onderzoek toont dat neurale stamcellen kunnen worden geïnduceerd om onderscheid te maken door veel van de stadia van oligodendrocytic afkomst ontwikkeling (progenitor-afgeleide oligodendrocyten, BOB). We cultuur neurale stamcellen in DMEM-F12 serumvrije media supaangevuld met basic fibroblast growth factor (bFGF), van bloedplaatjes afkomstige groeifactor (PDGF-AA), Sonic hedgehog (Shh), neurotrofische factor 3 (NT-3), N-2 en triiodothyronine (T3). De gekweekte cellen worden gepasseerd op 2.5e6 cellen per 75cm kolven ongeveer om de zeven dagen. Met deze omstandigheden is de meerderheid van de cellen in cultuur te behouden morfologie wordt gekenmerkt door een aantal processen en een automatische markers vooraf oligodendrocyt cellen, zoals A2B5 en O-4. Wanneer we de vier groeifactoren (GF) (bFGF, PDGF-AA, Shh, NT-3) verwijderen en toevoegen geconditioneerde media uit PDN, de cellen beginnen te meer processen en express markers specifiek voor oligodendrocyten differentiatie, als GalC en myeline verwerven basic protein (MBP). We fenotypische karakterisatie uitgevoerd met multicolor flowcytometrie unieke markers van oligodendrocyt identificeren.

Het is zeer belangrijk om de differentiatie starten vanaf een 70% -80% confluent neurale stamcellen kweek (Figuur 1A). Veel cellen zullen sterven na het wijzigen van het kweekmedium van stamvader tot oligo medium, omdat het voorziet in specifieke groeifactoren. Dit betekent dat de groei van neurale progenitorcellen niet gebonden aan een oligodendrocytic fenotype niet wordt ondersteund door het nieuwe medium (Figuur 1B). Incubatie in oligo medium + GF gedurende een week resulteerde in ee...

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

Primaire, menselijke foetale hersenen-afgeleide, multipotentiële progenitorcellen vermenigvuldigen<em&gt; In vitro</em&gt; Terwijl de kunnen differentiëren tot neuronen en astrocyten. Hieruit blijkt dat neurale stamcellen kunnen worden geïnduceerd door stadia van de oligodendrocytic lineage onderscheiden door conditionering met bepaalde groeifactoren.

Progenitor afgeleide oligodendrocyt Cultuur System van menselijke foetale hersenen

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

Research

JoVE Journal

Neuroscience

Generation of Neural Stem Cells from Discarded Human Fetal Cortical Tissue

Generatie van neurale stamcellen uit afgedankte menselijke foetale Cortical weefsel

Neural stem cells (NSCs) reside along the ventricular zone neuroepithelium during the development of the cortical plate. These early progenitors ...

Derivation of Enriched Oligodendrocyte Cultures and Oligodendrocyte/Neuron Myelinating Co-cultures from Post-natal Murine Tissues

Afleiding van Verrijkt oligodendrocyt Culturen en oligodendrocyt / Neuron Myelinating co-culturen van Post-natale Murine Tissues

Identifying the molecular mechanisms underlying OL development is not only critical to furthering our knowledge of OL biology, but also has implications ...

Isolation and Culture of Endothelial Cells from the Embryonic Forebrain

Isolatie en Cultuur van de endotheelcellen van de embryonale voorhersenen

Embryonic brain endothelial cells can serve as an important tool in the study of angiogenesis and neurovascular development and interactions. The two ...

A cGMP-applicable Expansion Method for Aggregates of Human Neural Stem and Progenitor Cells Derived From Pluripotent Stem Cells or Fetal Brain Tissue

Een cGMP-toepassing Uitbreiding Methode voor Aggregaten van menselijke neurale stamcellen en stamcellen afgeleid uit pluripotente stamcellen of foetaal hersenweefsel

A cell expansion technique to amass large numbers of cells from a single specimen for research experiments and clinical trials would greatly benefit the ...

Live Imaging of Primary Cerebral Cortex Cells Using a 2D Culture System

Live beeldvorming van primaire hersenschors cellen met behulp van een systeem van 2D cultuur

During cerebral cortex development, progenitor cells undergo several rounds of symmetric and asymmetric cell divisions to generate new progenitors or ...

Simple Generation of a High Yield Culture of Induced Neurons from Human Adult Skin Fibroblasts

Eenvoudige generatie van de cultuur van een hoog rendement van geïnduceerde neuronen van menselijke volwassen huid fibroblasten

Induced neurons (iNs), the product of somatic cells directly converted to neurons, are a way to obtain patient-derived neurons from tissue that is easily ...

Isolation of Cerebral Capillaries from Fresh Human Brain Tissue

Isolatie van cerebrale haarvaten van verse menselijk hersenweefsel

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

Migratie, Chemo-Attraction, en Co-Culture Assays voor menselijke stamcel-afgeleide endotheelcellen en GABAergic Neuronen

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

Meten en manipuleren van functioneel specifieke neurale paden in het menselijk motorsysteem met transcraniële magnetische stimulatie

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

Generatie oligodendrocyten en oligodendrocyte-geconditioneerde medium voor co-cultuurexperimenten

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