Name: a 5-mc dot blot assay quantifying the dna methylation level of chondrocyte dedifferentiation in vitro
Uploaded: 2017-05-17T13:00:00
Description: Watch this Scientific Journal Video about A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro at JoVE.com

Dette arbeidet ble støttet av følgende tilskudd: Natural Science Foundation of China (nr. 81572198; nr. 81260161; nr. 81000460); Natural Science Foundation i Guangdong-provinsen, Kina (nr. 2015A030313772); Kina Postdoktoravhengighetsfond finansiert prosjekt (nr. 2013M530385); The Medical Research Foundation i Guangdong-provinsen, Kina (nr. A2016314); Shenzhen Science and Technology Prosjekter (nr. JCYJ20160301111338144; No. JSGG20151030140325149; No. JSGG20140519105550503; No.GJHZ20130412153906739; No. JCYJ20140414170821160; No.JCYJ20140414170821200).

Denne studien ble godkjent av Human Ethics Committee i Shenzhen Second People&#39;s Hospital. 1. Human Articular Cartilage Tissue Collection og Chondrocyte Culture <ol><li> Fremstilling av materialer <ol><li> Forbered Dulbeccos modifiserte eagle medium (DMEM) medium supplert med 10% føtalt bovint serum og 1% penicillin streptomycin. Klargjør 1 mg / ml kollagenase II, 0,25% trypsin-EDTA, fosfatbuffert saltvann (PBS) og en cellefilter (40 μm nylon). </li></ol></li><li> <s...

<ol>
	<li>Brittberg, 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=Treatment+of+deep+cartilage+defects+in+the+knee+with+autologous+chondrocyte+transplantation.">Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation.</a> N Engl J Med. 331, 889-895 (1994).</li><li>Pareek, 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=Long-Term+Outcomes+After+Autologous+Chondrocyte+Implantation:+A+Systematic+Review+at+Mean+Follow-Up+of+11.4+Years.">Long-Term Outcomes After Autologous Chondrocyte Implantation: A Systematic Review at Mean Follow-Up of 11.4 Years.</a> Cartilage. 7 (4), 298-308 (2016).</li><li>Duan, 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=Cytokine+networking+of+chondrocyte+dedifferentiation+in+vitro+and+its+implications+for+cell-based+cartilage+therapy.">Cytokine networking of chondrocyte dedifferentiation in vitro and its implications for cell-based cartilage therapy.</a> Am J Transl Res. 7 (2), 194-208 (2015).</li><li>Ma, 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=Gene+expression+profiling+of+dedifferentiated+human+articular+chondrocytes+in+monolayer+culture.">Gene expression profiling of dedifferentiated human articular chondrocytes in monolayer culture.</a> Osteoarthritis Cartilage. 21 (4), 599-603 (2013).</li><li>Duan, L., Liang, Y., Ma, B., Zhu, W., Wang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epigenetic+regulation+in+chondrocyte+phenotype+maintenance+for+cell-based+cartilage+repair.">Epigenetic regulation in chondrocyte phenotype maintenance for cell-based cartilage repair.</a> Am J Transl Res. 7 (11), 2127-2140 (2015).</li><li>Duan, 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=DNA+methylation+profiling+in+chondrocyte+dedifferentiation+in+vitro.">DNA methylation profiling in chondrocyte dedifferentiation in vitro.</a> J Cell Physiol. , (2016).</li><li>Bhat, 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=DNA+methylation+detection+at+single+base+resolution+using+targeted+next+generation+bisulfite+sequencing+and+cross+validation+using+capillary+sequencing.">DNA methylation detection at single base resolution using targeted next generation bisulfite sequencing and cross validation using capillary sequencing.</a> Gene. , (2016).</li><li>Shi, X. W., 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=Exploring+Genome-wide+DNA+Methylation+Profiles+Altered+in+Kashin-Beck+Disease+Using+Infinium+Human+Methylation+450+Bead+Chips.">Exploring Genome-wide DNA Methylation Profiles Altered in Kashin-Beck Disease Using Infinium Human Methylation 450 Bead Chips.</a> Biomed Environ Sci. 29 (7), 539-543 (2016).</li><li>Li, X. 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=Optimization+of+an+HPLC+Method+for+Determining+the+Genomic+Methylation+Levels+of+Taxus+Cells.">Optimization of an HPLC Method for Determining the Genomic Methylation Levels of Taxus Cells.</a> J Chromatogr Sci. 54 (2), 200-205 (2016).</li><li>Maghbooli, Z., 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=Global+DNA+methylation+as+a+possible+biomarker+fordiabetic+retinopathy.">Global DNA methylation as a possible biomarker fordiabetic retinopathy.</a> Diabetes Metab Res Rev. 31 (2), 183-189 (2015).</li><li>Barlic, A., Drobnic, M., Malicev, E., Kregar-Velikonja, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+analysis+of+gene+expression+in+human+articular+chondrocytes+assigned+for+autologous+implantation.">Quantitative analysis of gene expression in human articular chondrocytes assigned for autologous implantation.</a> J Orthop Res. 26 (6), 847-853 (2008).</li><li>Legendre, F., 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=Enhanced+hyaline+cartilage+matrix+synthesis+in+collagen+sponge+scaffolds+by+using+siRNA+to+stabilizechondrocytes+phenotype+cultured+with+bone+morphogenetic+protein-2+under+hypoxia.">Enhanced hyaline cartilage matrix synthesis in collagen sponge scaffolds by using siRNA to stabilizechondrocytes phenotype cultured with bone morphogenetic protein-2 under hypoxia.</a> Tissue Eng Part C Methods. 19 (7), 550-567 (2013).</li><li>Niethammer, T. R., 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=Analysis+of+the+autologous+chondrocyte+quality+of+matrix-based+autologous+chondrocyte+implantation+in+the+knee+joint.">Analysis of the autologous chondrocyte quality of matrix-based autologous chondrocyte implantation in the knee joint.</a> Int Orthop. 40 (1), 205-212 (2016).</li><li>Hayatsu, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+bisulfite+genomic+sequencing+used+in+the+analysis+of+epigenetic+states,+a+technique+in+the+emerging+environmental+genotoxicology+research.">The bisulfite genomic sequencing used in the analysis of epigenetic states, a technique in the emerging environmental genotoxicology research.</a> Mutat Res. 659 (1-2), 77-82 (2008).</li><li>Haque, N., Nishiguchi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bisulfite+sequencing+for+cytosine-methylation+analysis+in+plants.">Bisulfite sequencing for cytosine-methylation analysis in plants.</a> Methods Mol Biol. 744, 187-197 (2011).</li><li>Ananiev, G. 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=Optical+mapping+discerns+genome+wide+DNA+methylation+profiles.">Optical mapping discerns genome wide DNA methylation profiles.</a> BMC Mol Biol. 9, 68 (2008).</li></ol>

Chondrocyt dedifferentiation in vitro kompromitterer svært utfallet av ACI i behandlingen av brusk defekt reparasjon 11 , 12 . For å optimalisere ACI-utfallet er det avgjørende å unngå bruk av dedifferentierte kondrocytter 13 . Studier har antydet at generelt DNA-metyleringsnivå er assosiert med omfanget av kondrocyt dedifferensiering 4 , 6 . Det er således avgjør...

Autolog chondrocytimplantasjon (ACI) er en relativt ny, state-of-the-art prosedyre for behandling av leddbruskdefekter 1 , 2 . En av de avgjørende trinnene i ACI er amplifisering av kondrocytter via monolagskultur in vitro . Under amplifisering, taper hyalinkondrocytterene lett sin fenotype og blir dedifferentiert, noe som er uønsket for ACI-behandling 3 , 4 . For å optimalisere utfallet av ACI-behandling, bør omfanget av kondrocyt dedifferentiering bestemmes før genplantning. Det er viktig å etablere en økonomisk og rask måte å bestemme kondrocytternes status på. Foreningen mellom DNA-metylering og kondrocyt dedifferentiering har nylig tiltrukket seg mye oppmerksomhet 4 , 5 , 6 . DNA-metylering er en prosess av whI metylgrupper tilsettes DNA, noe som resulterer i omdannelse av cytosinrester til 5-metylcytosin (5-mC). For å belyse biologien av DNA-metylering i kondrocyt dedifferentiering, er det første trinnet å evaluere DNA-metyleringsnivået av kondrocytter, som hittil har vist seg å være utfordrende. Bisulfitt genomisk sekvensering er den mest brukte teknikken for å analysere DNA-metylering 7 , 8 . I denne analysen forårsaker bisulfitt-omdannelse DNA-nedbrytning, og således må det tilveiebringes en vesentlig mengde prøve for analysen. Dessuten har høyytelsesvæskekromatografi (HPLC) blitt brukt til å kvantifisere globale DNA-metyleringsnivåer 9 , 10 . HPLC-analyse krever imidlertid genomisk DNA-fordøyelse. Videre kreves avanserte og dyre eksperimentelle instrumenter. Derfor, i tillegg til den høye prisen, er disse eksperimentelle prosedyrene tidsforstyrrelseruming. Anti-5-mC antistoffer har nå blitt kommersielt tilgjengelige, noe som har skapt muligheten for immunblotting av 5-mC-holdig genomisk DNA fra komplekse genomer. I denne rapporten ekstrahente vi genomisk DNA fra kondrocyter dyrket i en serie av monolagskulturer. Vi brukte en dot blot-analyse for å evaluere 5-mC innholdet i humane kondrocyter med forskjellige antall passasjer. Vi fant at 5-mC innhold ble økt i svært dedifferentierte kondrocytter sammenlignet med kondrocytter med lavverdig dedifferentiering. I tillegg identifiserte vi et forhold mellom dedifferentiation status og 5-mC nivåer. Endelig rapporterte vi at endringene i 5-mC innhold var assosiert med kondrocyt fenotypen. Derfor er 5-mC dot blot-analysen en pålitelig, enkel og rask metode for å oppdage DNA-metyleringsnivået i kondrocytter.

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			<td>Reagents</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM&#160;&#160;</td>
			<td>Gibco Inc.</td>
			<td>11965&#8211;092</td>
			<td>Warm in 37 &#176;C water bath before use</td>
		</tr>
		<tr>
			<td rowspan="2">Phosphate-Buffered Saline(PBS)</td>
			<td rowspan="2">HyClone Inc.</td>
			<td rowspan="2">SH30256.01B</td>
			<td>D-PBS, free of&#160;</td>
		</tr>
		<tr>
			<td>Ca2+/Mg2+</td>
		</tr>
		<tr>
			<td>FBS</td>
			<td>Gibco Inc.</td>
			<td>10099-141</td>
			<td></td>
		</tr>
		<tr>
			<td>0.25%Trypsin/EDTA</td>
			<td>Gibco Inc.</td>
			<td>25200-056</td>
			<td></td>
		</tr>
		<tr>
			<td>1%Penicillin-Streptomycin</td>
			<td>Gibco Inc.</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Chloroform</td>
			<td>Mallinckrodt</td>
			<td>4440</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoamyl Alcohol</td>
			<td>Sigma</td>
			<td>I-3643</td>
			<td></td>
		</tr>
		<tr>
			<td>Phenol</td>
			<td>Gibco BRL</td>
			<td>15513-039</td>
			<td></td>
		</tr>
		<tr>
			<td>Proteinase K</td>
			<td>Gibco BRL</td>
			<td>24568-2</td>
			<td></td>
		</tr>
		<tr>
			<td>TAE buffer&#160;</td>
			<td>Bio Whittaker</td>
			<td>16-011V</td>
			<td></td>
		</tr>
		<tr>
			<td>Distilled Water</td>
			<td>Gibco BRL</td>
			<td>15230-170</td>
			<td></td>
		</tr>
		<tr>
			<td>1 M Tris-HCl</td>
			<td>Biosharp Inc.</td>
			<td>BL514A</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween20</td>
			<td>Biotopped Inc.</td>
			<td>C58H114O26</td>
			<td></td>
		</tr>
		<tr>
			<td>BSA</td>
			<td>Proliant Inc.</td>
			<td>68700</td>
			<td></td>
		</tr>
		<tr>
			<td>Collagenase, Type II</td>
			<td>Sigma-Aldrich</td>
			<td>C6885</td>
			<td></td>
		</tr>
		<tr>
			<td>Names&#160;&#160;&#160;&#160;&#160;</td>
			<td>Company&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td>Catalog number&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td>Comments</td>
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			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cell Strainer(40&#956;m nylon&#65289;</td>
			<td>FALCON Inc.</td>
			<td>352340</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemocytometer</td>
			<td>ISOLAB Inc.</td>
			<td>075.03.001</td>
			<td></td>
		</tr>
		<tr>
			<td>Falcon 100 mm&#160; dish</td>
			<td>Corning</td>
			<td>353003</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge Tubes</td>
			<td>TPP AG</td>
			<td>91050</td>
			<td>Gamma-sterilized</td>
		</tr>
		<tr>
			<td>High-speed centrifuge</td>
			<td>Eppendorf</td>
			<td>5804R</td>
			<td></td>
		</tr>
		<tr>
			<td>ThermoMixer</td>
			<td>MIULAB</td>
			<td>MTH-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbon dioxide cell incubator</td>
			<td>Thermo scientific</td>
			<td>3111</td>
			<td></td>
		</tr>
		<tr>
			<td>Chemi-imaging Analyse System</td>
			<td>UVITEC Cambridge</td>
			<td>ALLIANCE</td>
			<td></td>
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a 5-mc dot blot assay quantifying the dna methylation level of chondrocyte dedifferentiation in vitro

Den dedifferentiering av hyalinkondrocytter til fibroblastiske kondrocytter følger ofte monolag-ekspansjon av kondrocytter in vitro . Det globale DNA-metyleringsnivået for kondrocyter anses å være en egnet biomarkør for tap av kondrocytfenotypen. Imidlertid kan resultater basert på forskjellige eksperimentelle metoder være inkonsekvente. Derfor er det viktig å etablere en presis, enkel og rask metode for å kvantifisere globale DNA-metyleringsnivåer under kondrocyt dedifferensiering. Aktuelle genom-brede metyleringsanalyse teknikker bygger i stor grad på bisulfitt genomisk sekvensering. På grunn av DNA-nedbrytning under bisulfittomdannelse krever disse metodene typisk et stort prøvevolum. Andre metoder som brukes til å kvantifisere globale DNA-metyleringsnivåer inkluderer høyytelsesvæskekromatografi (HPLC). HPLC krever imidlertid fullstendig fordøyelse av genomisk DNA. I tillegg er forbudt høye kostnader for HPLC iStruments begrenser HPLCs bredere anvendelse. I denne studien ble genomisk DNA (gDNA) ekstrahert fra humane kondrocyter dyrket med varierende antall passasjer. GDNA-metyleringsnivået ble detektert under anvendelse av en metyleringsspesifikk dot-blot-analyse. I denne dot blot-tilnærmingen ble en gDNA-blanding inneholdende det metylerte DNA som skal detekteres, spottet direkte på en N + -membran som en prikk inne i et tidligere tegnet sirkulært malemønster. Sammenlignet med andre gelelektroforese-baserte blotting-tilnærminger og andre komplekse blottingprosedyrer sparer dot blot-metoden betydelig tid. I tillegg kan dotblots detektere det totale DNA-metyleringsnivå ved bruk av et kommersielt tilgjengelig 5-mC antistoff. Vi fant at DNA-metyleringsnivået varierte mellom monolag-subkulturer, og kunne derfor spille en nøkkelrolle i kondondrocyt dedifferentiering. 5-mC dot blot er en pålitelig, enkel og rask metode for å oppdage det generelle DNA-metyleringsnivået for å evaluereE kondrocyt fenotype.

Chondrocytter ble dyrket i et monolag opp til passasje 6 (P6). Kondrocytter viste progressive fenotypiske endringer med suksessive passasjer av monolagskulturen. P0-kondrocytemorfologien var rund, mens cellene ble svært klemt og flatet med suksessive passasjer opp til P6 ( Figur 1 ). Denne morfologiske forandringen er typisk for kondrocyt dedifferentieringsprosessen. I mellomtiden indikerte resultatene av varmekartet at det generelle metyleringsnivået for CpG-steder ø...

Watch this Scientific Journal Video about A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro at JoVE.com

A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro

Vi presenterer en metode for å kvantifisere DNA-metylering basert på 5-metylcytosin (5-mC) dot blot. Vi bestemte oss for 5-mC nivåene under chondrocyt dedifferentiering. Denne enkle teknikken kan brukes til å raskt bestemme kondrocytfenotypen ved ACI-behandling.

En 5-mC prikkblot-analyse Kvantifisering av DNA-metyleringsnivået av kondrocyt-diffusjon<em&gt; I Vitro</em

The dedifferentiation of hyaline chondrocytes into fibroblastic chondrocytes often accompanies monolayer expansion of chondrocytes in vitro. The global ...

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A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro

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