Name: from mefs to matrigel 2: splitting hescs from mefs onto matrigel
Uploaded: 2008-06-09T18:48:00
Description: Watch this Scientific Journal Video about From MEFs to Matrigel 2: Splitting hESCs from MEFs onto Matrigel at JoVE.com

<p> Menselijke embryonale stamcellen studies in de Teitell lab worden ondersteund door een Californische Instituut voor Regeneratieve Geneeskunde (CIRM) Seed Grant RS1-00313. Wij danken de leden van de globale Centrum voor Regeneratieve Geneeskunde en Stamcelonderzoek aan de UCLA, in het bijzonder Dr Amander Clark, Dr Jerome Zack, en leden van de UCLA Broad Institute Stem Cell Core Facility voor hun steun aan onze studies. </p>

<p class="jove_title"> hESC cultuur dagelijks onderhoud </p><ol><li> hESC cultuur media moeten worden veranderd om de 24 uur. Van de ES media voorraad fles bewaard bij 4 ° C, neemt de hoeveelheid oplossing nodig is (~ 20 ml per 6-well plaat), plaats in een 50 ml falcon buis, en warm tot 37 ° C in een waterbad. Zodra de media is opgewarmd, voeg bFGF opgeslagen bij 4 ° C tot een uiteindelijke concentratie van 10ng/ml. Plaats de bFGF voorraad bij 4 ° C direct na gebruik! </li><li> Alle uit de 6-well platen, maar ~ 500μl van de media. Zorg erv...

<ol>
	<li>Thomson, J. a. m. e. s. A., Itskovitz-Eldor, J. o. s. e. p. h., Shapiro, S. a. n. d. e. r. S., Waknitz, M. i. c. h. e. l. l. e. A., Swiergiel, J. e. n. n. i. f. e. r. J., Marshall, V. i. v. i. e. n. n. e. S., Jones, J. e. f. f. r. e. y. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Embryonic+Stem+Cell+Lines+Derived+from+Human+Blastocysts++.">Embryonic Stem Cell Lines Derived from Human Blastocysts  .</a> <em style='font-style: italic'>Science</em>. <b>282</b>, 1145-1147 (1998).</li><li>Xu, C. h. u. n. h. u. i., Inokuma, M. a. r. g. a. r. e. t. S., Denham, J. e. r. r. o. d., Golds, K. a. t. h. a. l. e. e. n., Kundu, P. r. a. t. i. m. a., Gold, J. o. s. e. p. h. D., Carpenter, M. e. l. i. s. s. a. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feeder-free+growth+of+undifferentiated+human+embryonic+stem+cells.">Feeder-free growth of undifferentiated human embryonic stem cells.</a> <em style='font-style: italic'>Nature Biotechnology</em>. <b>19</b>, 971-974 (2001).</li></ol>

<p> Deze video laat zien hoe passage hESC &#39;s van MEF platen om feeder celvrije Matrigel platen. Merk op dat de kolonie dichtheid op Matrigel hoger is dan de dichtheid van de kolonies door de gebruikelijke splitsen op MEFs. Immunofluorescentiekleuring en microscopie of flowcytometrie voor hESC pluripotentie markers, zoals Oct-4 en SSEA-4, zijn nodig om het onderhoud van hESC &#39;s in een ongedifferentieerde staat te bevestigen in feeder-vrije kweekomstandigheden. </p>

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<td>Gibco</td>
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<td>Reagent</td>
<td>Gibco</td>
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<td>Gibco</td>
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<td>Penicillin / Streptomycin </td>
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<td>&nbsp;</td>
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<td>FITCI-conjugated antirabbit IgG</td>
<td>Reagent</td>
<td>Jackson ImmunoResearch Laboratories. Inc.</td>
<td>715-095-152</td>
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from mefs to matrigel 2: splitting hescs from mefs onto matrigel

<p class="jove_content">Deze video laat zien hoe menselijke embryonale stamcellen (hESC &#39;s) groeien op muis embryonale fibroblasten (MEF) feeder-cellen, hoe passage hESC&#39; s van MEF platen om feeder celvrije Matrigel platen.</p>

Watch this Scientific Journal Video about From MEFs to Matrigel 2: Splitting hESCs from MEFs onto Matrigel at JoVE.com

From MEFs to Matrigel 2: Splitting hESCs from MEFs onto Matrigel

<p class="jove_content">Deze video laat zien hoe het handhaven van de groei van menselijke embryonale stamcellen (hESC &#39;s) in de feeder cel-vrije omstandigheden en hoe je continu passage hESC&#39; s in de feeder cel-vrije omstandigheden. Bevestiging van hESC pluripotentie gegroeid in feeder celvrije voorwaarden door immunofluorescentie microscopie is ook aangetoond. Deel 2 van 3.</p>

Van MEF tot Matrigel 2: Splitsen van hESC &#39;s van MEFs op Matrigel

This video demonstrates how to grow human embryonic stem cells (hESCs) on mouse embryonic fibroblast (MEF) feeder cells, how to passage hESCs from MEF ...

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Generation of a homogeneous and abundant population of skeletal muscle cells from human embryonic stem cells (hESCs) is a requirement for cell-based ...

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