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>在Teitell实验室的人类胚胎干细胞研究是由加州再生医学研究所（CIRM）种子批RS1 - 00313的支持。我们感谢的再生医学和干细胞研究的加州大学洛杉矶分校，特别是博士Amander克拉克，博士杰罗姆扎克，和加州大学洛杉矶分校的Broad研究所干细胞核心设施为支持我们的研究成员的广泛中心的成员。 </p>

<p class="jove_title">人类胚胎干细胞文化的日常维护</p><ol><li>人类胚胎干细胞的培养基必须24小时更换一次。从ES传媒股票在4 ° C储存瓶，拿出解决方案所需（〜20毫升每6孔板），在一个50毫升猎鹰管地方，和温暖的37℃水浴中。一旦媒体被加热，添加碱性成纤维细胞生长因子保存在4 ° C的终浓度在10ng/ml。在4 ° C，使用后立即把碱性成纤维细胞生长因子股票！ </li><li>从6孔板删除，但〜媒体500μL。确保摇动培养皿暂停去?...

<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>本视频演示了如何通过胚胎干细胞MEF板料器无细胞基质胶板。请注意，对基质胶的殖民地密度高于MEFs通常分裂菌落密度。人类胚胎干细胞全能性的标志物，如Oct - 4和SSEA - 4，免疫荧光染色和显微镜或流式细胞仪需要在馈线的文化条件，以确认人类胚胎干细胞保持在未分化状态。 </p>

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<td>Knockout Serum Replacer (KSR)</td>
<td>Reagent</td>
<td>Gibco</td>
<td>10828-028</td>
<td>&nbsp;</td>
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<td>DMEM/F12</td>
<td>Reagent</td>
<td>Gibco</td>
<td>11330-057</td>
<td>&nbsp;</td>
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<td> Non-essential Amino Acids</td>
<td>Reagent</td>
<td>Gibco</td>
<td>11140-050</td>
<td>&nbsp;</td>
<tr>
<td>GlutaMax </td>
<td>Reagent</td>
<td>Gibco</td>
<td>35050-061</td>
<td>&nbsp;</td>
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<td>DMEM</td>
<td>Reagent</td>
<td>Gibco</td>
<td>11995-065</td>
<td>&nbsp;</td>
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<td>FBS</td>
<td>Reagent</td>
<td>Clontech</td>
<td>631107</td>
<td>&nbsp;</td>
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<td>Reagent</td>
<td>Gibco</td>
<td>25030-081</td>
<td>&nbsp;</td>
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<td>Reagent</td>
<td>Fisher</td>
<td>BP176-100</td>
<td>&nbsp;</td>
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<td>bFGF</td>
<td>Reagent</td>
<td>R&D Systems</td>
<td>233-FB-025</td>
<td>&nbsp;</td>
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<td>Collagenase IV</td>
<td>Reagent</td>
<td>Gibco</td>
<td>17104-019</td>
<td>&nbsp;</td>
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<td>Dispase</td>
<td>Reagent</td>
<td>Stem Cell Technologies</td>
<td>17105-041</td>
<td>&nbsp;</td>
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<td>Penicillin / Streptomycin </td>
<td>Reagent</td>
<td>Gibco</td>
<td>15140-122</td>
<td>&nbsp;</td>
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<td>Gelatin</td>
<td>Reagent</td>
<td>Chemicon</td>
<td>ES-006-B</td>
<td>&nbsp;</td>
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<td>Matrigel</td>
<td>Reagent</td>
<td>BD Biosciences</td>
<td>354277</td>
<td>&nbsp;</td>
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<td>Oct-4 antibody</td>
<td>Reagent</td>
<td>Santa Cruz Biotechnology</td>
<td>SC-9081</td>
<td>&nbsp;</td>
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<td>anti-h/mSSEA-4 Phycoerythrin Conjugated Mouse IgG3</td>
<td>Reagent</td>
<td>R&D Systems</td>
<td>FAB1435P</td>
<td>&nbsp;</td>
<tr>
<td>FITCI-conjugated antirabbit IgG</td>
<td>Reagent</td>
<td>Jackson ImmunoResearch Laboratories. Inc.</td>
<td>715-095-152</td>
<td>&nbsp;</td>		</tbody>
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from mefs to matrigel 2: splitting hescs from mefs onto matrigel

<p class="jove_content">本视频演示了如何生长在人类胚胎干细胞（胚胎干细胞）的小鼠胚胎成纤维细胞（MEF）饲养层细胞，如何通过MEF的板人类胚胎干细胞馈线无细胞基质胶板。</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">本视频演示了如何维持人类胚胎干细胞（胚胎干细胞）在馈线无细胞的条件和如何在馈线不断通过人类胚胎干细胞的无细胞条件的增长。在馈线无细胞的条件下，免疫荧光显微镜确认的人类胚胎干细胞的全能性增长也表明。</p>

从MEFs基质胶2：到基质胶从MEFs分裂的胚胎干细胞

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

Research

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From MEFs to Matrigel I: Passaging hESCs in the Presence of MEFs

从MEFs基质胶我的MEFs传代的胚胎干细胞

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

科研

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Generating iPS Cells from MEFS through Forced Expression of Sox-2, Oct-4, c-Myc, and Klf4

从MEFS iPS细胞生成通过强制SOX - 2的表达，10月4，c - Myc基因和Klf4

Pluripotency can be induced in differentiated murine by viral transduction of Oct4, Sox2, Klf4, and c-Myc (Takahashi and Yamanaka, 2006;  Wernig, et al., ...

From MEFs to Matrigel 3: Passaging hESCs from Matrigel onto Matrigel

从MEFs基质胶3：传代的胚胎干细胞从基质胶到基质胶

This video demonstrates how to maintain the growth of human embryonic stem cells (hESCs) in feeder cell-free conditions and how to continuously passage ...

Cargo Loading onto Kinesin Powered Molecular Shuttles

货物装载到驱动蛋白供电分子梭

Cells have evolved sophisticated molecular machinery, such as kinesin motor proteins and microtubule filaments, to support active intracellular transport ...

An Introduction to Worm Lab: from Culturing Worms to Mutagenesis

蠕虫病毒实验室介绍：从培养蠕虫突变

This protocol describes procedures to maintain nematodes in the laboratory and how to mutagenize them using two alternative methods: ethyl methane ...

A Matrigel-Based Tube Formation Assay to Assess the Vasculogenic Activity of Tumor Cells

基于一个基质胶管形成实验，以评估对肿瘤细胞的血管生成活性

Over the past several decades, a tube formation assay using growth factor-reduced Matrigel has been typically employed to demonstrate the angiogenic ...

Generation of Myospheres From hESCs by Epigenetic Reprogramming

Myospheres的产生从人类胚胎干细胞通过表观遗传修饰

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

<em>Xenopus laevis</em> as a Model to Identify Translation Impairment

Xenopus laevis as a Model to Identify Translation Impairment

<em>非洲爪蟾</em>作为一种模型来确定翻译减值

Protein synthesis is a fundamental process to gene expression impacting diverse biological processes notably adaptation to environmental conditions. The ...

Using Caco-2 Cells to Study Lipid Transport by the Intestine

采用Caco-2细胞研究脂质运输由肠

Studies of dietary fat absorption are generally conducted by using an animal model equipped with a lymph cannula. Although this animal model is widely ...

Magnetic-Activated Cell Sorting Strategies to Isolate and Purify Synovial Fluid-Derived Mesenchymal Stem Cells from a Rabbit Model

从兔模型中分离纯化滑液源性间充质干细胞的磁活化细胞分选策略

Mesenchymal stem cells (MSCs) are the main cell source for cell-based therapy. MSCs from articular cavity synovial fluid could potentially be used for ...