Name: preparation of primary mixed glial cultures from adult mouse spinal cord tissue
Uploaded: 2016-11-19T14:00:00
Description: Watch this Scientific Journal Video about Preparation of Primary Mixed Glial Cultures from Adult Mouse Spinal Cord Tissue at JoVE.com

Adult spinal cord mixed cultures were first developed using rats with grant support from an NIH/NIDA R01 award (PI De Leo) and an NIH T32 training grant (PI Green). These methods were further adapted for mouse spinal cords with support from the following grants: NIH/NIDA 5K01DA023503 (PI Cao), NIH/NINDS 5R21NS066130 (PI Cao), and NIH/NIGMS P20GM103643 (PI Meng). The authors would also like to thank Dr. Alejandro M. S. Mayer, Department of Pharmacology, Chicago College of Osteopathic Medicine, for his technical help in obtaining microglia-rich mixed glial cells.

Følgende protokol blev godkendt af Institutional Animal Care og brug Udvalg (IACUC) på University of New England. Følgende protokol er til at forberede gliale kulturer fra 4 voksne mus rygmarv. 1. Udarbejdelse af løsninger i en kultur Hood under aseptiske forhold <ol><li> Forbered dyrkningsmedier: komplet Dulbeccos modifikation af Eagles medium (cDMEM) indeholdende DMEM (med 4,5 g / l glucose), 10% kalvefosterserum (FBS), 2 mM L-glutamin, 100 IE / ml penicillin, 100 mg / ml streptomyci...

<ol>
	<li>Gaskin, D. J., Richard, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+economic+costs+of+pain+in+the+United+States.">The economic costs of pain in the United States.</a> J Pain. 13 (8), 715-724 (2012).</li><li>Ji, R. R., Berta, T., Nedergaard, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glia+and+pain:+Is+chronic+pain+a+gliopathy?.">Glia and pain: Is chronic pain a gliopathy?.</a> Pain. 154 (01), S10-S28 (2013).</li><li>Ni, M., Aschner, M., Maines, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neonatal+rat+primary+icroglia:+isolation,+culturing+and+selected+applications.">Neonatal rat primary icroglia: isolation, culturing and selected applications.</a> Curr Protoc Toxicol. , (2010).</li><li>Cao, L., Fei, L., Chang, T. T., DeLeo, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+interleukin-1beta+by+interleukin-4+in+lipopolysaccharide-treated+mixed+glial+cultures:+microglial-dependent+effects.">Induction of interleukin-1beta by interleukin-4 in lipopolysaccharide-treated mixed glial cultures: microglial-dependent effects.</a> J Neurochem. 102 (2), 408-419 (2007).</li><li>Malon, J. T., Maddula, S., Bell, H., Cao, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+calcitonin+gene-related+peptide+and+CCL2+production+in+CD40-mediated+behavioral+hypersensitivity+in+a+model+of+neuropathic+pain.">Involvement of calcitonin gene-related peptide and CCL2 production in CD40-mediated behavioral hypersensitivity in a model of neuropathic pain.</a> Neuron Glia Biol. 7 (2-4), 117-128 (2011).</li><li>Blasi, E., Barluzzi, R., Bocchini, V., Mazzolla, R., Bistoni, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immortalization+of+murine+microglial+cells+by+a+v-raf+/+v-myc+carrying+retrovirus.">Immortalization of murine microglial cells by a v-raf / v-myc carrying retrovirus.</a> J Neuroimmunol. 27 (2), 229-237 (1990).</li><li>Bocchini, V., 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=An+immortalized+cell+line+expresses+properties+of+activated+microglial+cells.">An immortalized cell line expresses properties of activated microglial cells.</a> J Neurosci Res. 31 (4), 616-621 (1992).</li><li>Alliot, F. O., Marty, M. C., Cambier, D., Pessac, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+spontaneously+immortalized+mouse+microglial+cell+line+expressing+CD4.">A spontaneously immortalized mouse microglial cell line expressing CD4.</a> Dev Brain Res. 95 (1), 140-143 (1996).</li><li>Alliot, F. O., Pessac, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Astrocytic+cell+clones+derived+from+established+cultures+of+8-day+postnatal+mouse+cerebella.">Astrocytic cell clones derived from established cultures of 8-day postnatal mouse cerebella.</a> Brain Res. 306 (1-2), 283-291 (1984).</li><li>Malon, J. T., 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=Microglial+content-dependent+inhibitory+effects+of+calcitonin+gene-related+peptide+(CGRP)+on+murine+retroviral+infection+of+glial+cells.">Microglial content-dependent inhibitory effects of calcitonin gene-related peptide (CGRP) on murine retroviral infection of glial cells.</a> J Neuroimmunol. 279, 64-70 (2015).</li><li>Cao, L., DeLeo, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CNS+Infiltrating+CD4(+)+T+lymphocytes+Contribute+to+Murine+Spinal+Nerve+Transection-Induced+Neuropathic+Pain.">CNS Infiltrating CD4(+) T lymphocytes Contribute to Murine Spinal Nerve Transection-Induced Neuropathic Pain.</a> Eur J Immunol. 38 (2), 448-458 (2008).</li><li>Mayer, A. M., 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=Effect+of+a+short-term+in+vitro+exposure+to+the+marine+toxin+domoic+acid+on+viability,+tumor+necrosis+factor-alpha,+matrix+metalloproteinase-9+and+superoxide+anion+release+by+rat+neonatal+microglia.">Effect of a short-term in vitro exposure to the marine toxin domoic acid on viability, tumor necrosis factor-alpha, matrix metalloproteinase-9 and superoxide anion release by rat neonatal microglia.</a> BMC Pharmacol. 1, 7 (2001).</li><li>Lawson, L. J., Perry, V. H., Dri, P., Gordon, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneity+in+the+distribution+and+morphology+of+microglia+in+the+normal+adult+mouse+brain.">Heterogeneity in the distribution and morphology of microglia in the normal adult mouse brain.</a> Neuroscience. 39 (1), 151-170 (1990).</li><li>Rock, R. B., 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=Role+of+Microglia+in+Central+Nervous+System+Infections.">Role of Microglia in Central Nervous System Infections.</a> Clin Microbiol Rev. 17 (4), 942-964 (2004).</li><li>Yang, I., Han, S. J., Kaur, G., Crane, C., Parsa, A. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Role+of+Microglia+in+Central+Nervous+System+Immunity+and+Glioma+Immunology.">The Role of Microglia in Central Nervous System Immunity and Glioma Immunology.</a> J Clin Neurosci. 17 (1), 6-10 (2010).</li></ol>

Det er afgørende at udføre alle trin-herunder medierne skiftende tidsplan-på en sammenhængende måde for at opnå reproducerbare resultater mellem eksperimenter. Følgende trin er særligt kritiske i at opnå fremragende kvalitet voksen blandet glia. Den enzymatiske fordøjelse er et vigtigt aspekt af denne protokol. Det er afgørende, at de vævsstykker er godt fordøjet for at opnå en enkelt cellesuspension. Dog vil over-fordøjelse resultere i væsentligt færre celler. Hver lab beho...

Kroniske smerter er et alvorligt sundhedsproblem, der rammer cirka 100 millioner voksne i USA, med en anslået årlig udgift på op til $ 635 1 mia. Voksende vidnesbyrd har indikeret et væsentligt bidrag rygmarv gliaceller i udviklingen af neuropatisk smerte, en af de mest ødelæggende former for kronisk smerte 2. En ordentlig in vitro kultursystem ville bidrage betydeligt i yderligere at undersøge den rolle, gliaceller på cellulært og molekylært niveau. I øjeblikket er de in vitro glia dyrkningssystemer anvendes af forskere omfatter hovedsagelig gliale celler afledt fra corticale væv af neonatale mus eller rotte hjerner og immortaliserede gliale cellelinier afledt fra neonatal mus eller rotte primære gliaceller. Neonatale celler indeholder et betydeligt antal udifferentierede celler, der kan være yderligere differentieret i gliaceller (hovedsagelig astrocytter og mikroglia), hvilket således tilvejebringer enbundant celler til eksperimentel brug 3. vores tidligere undersøgelse har imidlertid vist, at neonatal hjerne gliaceller reagerede signifikant forskelligt fra voksne rygmarven gliaceller efter lipopolysaccharid (LPS) stimulation. For eksempel, interleukin (IL) -4 de viste forøget inhiberende virkninger på LPS-induceret nitrogenoxid (NO) produktion i voksne rotter rygmarv glia sammenlignet med neonatal hjerne glia 4. Endvidere profiler af chemokin produktion ved stimulering af et neuropeptid, calcitonin gen-relateret peptid (CGRP), er ubestrideligt forskellige mellem neonatal musehjerne glia (metoder beskrevet i Ref. 5), og voksne mus rygmarv glia (figur 1). Etablerede cellelinjer er nemme at anvende og vedligeholde og kan give et stort antal celler i en kort periode. Generelt immortaliserede cellelinjer genereres enten ved hjælp af en virus-medieret immortalisering systemet (såsom udbredte mikroglial cellelinie BV2) 6, 7 eller efter the identifikation af spontan transformation (såsom astrocyt cellelinje C8-D1A og mikroglial cellelinie C8-B4) 8, 9 cellelinier er fremragende i at studere de molekylære karakteristika af astrocytter og mikroglia individuelt.; imidlertid opnået fra cellelinjer resultater kræver altid yderligere validering i primære celler eller i in vivo-betingelser. Det skal også bemærkes, at der ikke har været en rapport fra et glial cellelinie, som er afledt af en gnaver rygmarv glia. For at hjælpe undersøge betydningen af rygmarven gliaceller i udviklingen af neuropatisk smerte, en kultur, der er afledt af voksne mus rygmarven er blevet udviklet ved at tilpasse en tidligere rapporteret fremgangsmåde anvendes til at generere voksne rotter blandede gliale kulturer 4, 5 . musen rygmarv glial kultur blev yderligere forfinet nylig 10 og er beskrevet mere detaljeret i denne artikel. Voksen rygmarv blandet gliale kulturer cen skal etableres med en mus fra udvalgte stammer, som passer til behovene i den særlige undersøgelse, og cellerne kan opretholdes i kultur i op til 21 d sende initiering af kulturen. Denne metode kan anvendes i neuropatisk smerte studier, såvel som i undersøgelser af forskellige neurologiske sygdomme, som medfører patologiske ændringer i rygmarven, såsom amyotrofisk lateral sklerose (ALS), human immundefekt virus (HIV) -associeret sensorisk neuropati og multipel sklerose (MS).

<table border="0" cellpadding="0" cellspacing="0" width="2053">
	<tbody>
		<tr height="24">
			<td height="24" style="height:24px;width:489px;">Dulbecco&#8217;s modification of Eagle&#8217;s media (DMEM) with 4.5g/L glucose</td>
			<td style="width:277px;">Lonza</td>
			<td style="width:119px;">12-709F</td>
			<td style="width:1168px;">The cDMEM media is a standard, widely used culture media. Individual researcher can decide where to purchase the DMEM and all other component used to make cDMEM media.</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">L-Glutamine (100x)</td>
			<td style="width:277px;">Lonza</td>
			<td style="width:119px;">17-605E</td>
			<td>The L-glutamine is a standard, widely used component for various culture media. Individual researcher can decide where to purchase L-glutamine.</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:489px;">Antibiotic-Antimycotic Solution (100x)</td>
			<td style="width:277px;">Corning-Mediatech</td>
			<td style="width:119px;">30-004-CI</td>
			<td>This is a combination of penicillin, streptomycin and Amphotericin formulated to contain 10,000 units/ml penicillin G, 10 mg/ml streptomycin sulfate and 25 &#181;g/ml amphotericin B. Individual researcher can decide where to purchase individual component.</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">2-mercaptoethanol (2-ME)</td>
			<td style="width:277px;">Sigma-Aldrich</td>
			<td style="width:119px;">M3148</td>
			<td>A BioReagent, suitable for cell culture, molecular biology and electrophoresis.</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Papain dissociation system</td>
			<td style="width:277px;">Worthington Biochemical Corporation</td>
			<td>LK003150</td>
			<td>Individual components of this kit can be purchased separately.&#160;</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:489px;">Percoll</td>
			<td>GE Health Care</td>
			<td style="width:119px;">17-0891-01</td>
			<td>Percoll is sold as sterile solution. Undiluted Percoll can be re-autoclaved if needed.</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:489px;">Lab-line incubator/shaker&#160;</td>
			<td style="width:277px;">Barstead/Lab-line</td>
			<td style="width:119px;">MaxQ4000&#160;</td>
			<td>This is the incubator/shaker we have currently. Other types of shakers can be used instead.</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:489px;">Lipopolysaccharides, Salmonella Minnesota Re595&#160;</td>
			<td style="width:277px;">Sigma-Aldrich</td>
			<td style="width:119px;">L-9764</td>
			<td>Other strains of LPS can also induce glial responses. The magnitude of the responses may vary.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:489px;">Mouse IL-6 DuoSet ELISA&#160;</td>
			<td style="width:277px;">R&#38;D Systems</td>
			<td style="width:119px;">DY406</td>
			<td>Each individual Researcher can select the ELISA kit he or she prefers.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:489px;">Mouse TNF-alpha DuoSet ELISA</td>
			<td style="width:277px;">R&#38;D Systems</td>
			<td style="width:119px;">DY410</td>
			<td>Each individual Researcher can select the ELISA kit he or she prefers.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:489px;">Mouse IP-10 (CXCL10) DuoSet ELISA</td>
			<td style="width:277px;">R&#38;D Systems</td>
			<td style="width:119px;">DY466</td>
			<td>Each individual Researcher can select the ELISA kit he or she prefers.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:489px;">Mouse MCP-1 (CCL2) ELISA Opteia set</td>
			<td style="width:277px;">BD Biosciences</td>
			<td style="width:119px;">555260</td>
			<td>Each individual Researcher can select the ELISA kit he or she prefers.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:489px;">Mouse chemokine Q-Plex</td>
			<td style="width:277px;">Quansys Biosciences</td>
			<td style="width:119px;">120251MS</td>
			<td>This product was available for in-house assay at the time. Instead, we sent out samples to the manufacturer for analysis.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:489px;">APC-anti-mouse-CD45 (clone 30-F11)</td>
			<td style="width:277px;">eBioscience</td>
			<td style="width:119px;">17-0451</td>
			<td>Antibody of the same clone but from other vendors can be used.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:489px;">FITC-anti-mouse-CD11b (clone M1/70)</td>
			<td style="width:277px;">eBioscience</td>
			<td style="width:119px;">11-0112</td>
			<td>Antibody of the same clone but from other vendors can be used.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:489px;">Accuri C6 flow cytometer</td>
			<td style="width:277px;">BD Biosciences</td>
			<td style="width:119px;">BD Accuri C6&#160;</td>
			<td>Each individual Researcher can use any flow cytometer he or she prefers.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:489px;">FlowJo software</td>
			<td style="width:277px;">Tree Star, Inc.</td>
			<td style="width:119px;">FlowJo7.6.5</td>
			<td>Each individual Researcher can use any analysis software he or she prefers.</td>
		</tr>
	</tbody>
</table>

preparation of primary mixed glial cultures from adult mouse spinal cord tissue

Det er blevet godt accepteret, at rygmarv gliale reaktioner bidrager væsentligt til udviklingen af ​​neuropatisk smerte. Enorm oplysninger om gliale aktiviteter på de cellulære og molekylære niveau er opnået gennem in vitro cellekultursystemer. In vitro-systemer anvendes, omfatter hovedsagelig primære glia stammer fra neonatal hjerne kortikale væv og udødeliggjort cellelinjer. Imidlertid kan disse systemer ikke afspejler kendetegnene ved rygmarven gliaceller in vivo. For yderligere at undersøge rollerne for rygmarven gliaceller i udviklingen af perifer nerveskade-induceret neuropatisk smerte ved anvendelse af et dyrkningssystem, der bedre afspejler in vivo tilstand, har vores laboratorium udviklet en metode til at etablere primær rygmarv blandede gliaceller kulturer fra voksne mus. Kort fortalt rygmarv indsamlet fra voksne mus og behandles gennem papainspaltning efterfulgt af myelin fjernelse med et hulerity-gradient medium. Enkeltcellesuspensioner dyrkes i komplet Dulbeccos modificerede Eagle medium (cDMEM) suppleret med 2-mercaptoethanol (2-ME) ved 35,9 ° C. Disse dyrkningsbetingelser blev optimeret specielt til vækst af blandede gliaceller. Under disse betingelser er celler klar til at blive brugt til forsøg på mellem 12 - 14 d (celler er normalt i log-fase i denne periode) efter etableringen af ​​kulturen (D 0) og kan holdes i dyrkningsbetingelser op til D 21. dette dyrkningssystem kan anvendes til at undersøge responser af rygmarven gliaceller ved stimulering med forskellige stoffer og midler. Udover neuropatisk smerte, kan dette system anvendes til at studere gliale reaktioner i andre sygdomme, som medfører patologiske ændringer i rygmarven gliaceller.

Denne metode kan anvendes til fremstilling af blandede gliaceller fra både mus og rotter. Den gennemsnitlige totale celleantal pr brønd i en plade med 12 brønde på D 12 post-initiering af kulturen bør være relativt stabil, med omkring 100.000 celler per brønd når celler afledt fra muse rygmarv. Gliaceller opnået fra denne metode kan bruges i forsøg, der er designet til at undersøge voksne rygmarven gliale reaktioner efter indgivelse af stoffer og midler af interesse. F...

Watch this Scientific Journal Video about Preparation of Primary Mixed Glial Cultures from Adult Mouse Spinal Cord Tissue at JoVE.com

Preparation of Primary Mixed Glial Cultures from Adult Mouse Spinal Cord Tissue

Udviklingen af ​​neuropatisk smerte omfatter patologiske ændringer i rygmarven gliaceller. En pålidelig glia kultur-system stammer fra voksen rygmarv væv og designet til at studere disse celler in vitro mangler. Derfor viser vi her hvordan man etablerer primære blandede gliale kulturer fra voksne mus rygmarven væv.

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