Name: freezing human es cells
Uploaded: 2006-10-12T00:00:00
Description: Watch this Scientific Journal Video about Freezing Human ES Cells at JoVE.com

<ol><li>一个健康，成倍扩大培养，用PBS洗，以去除残留的媒体，否则可能淬火胰蛋白酶反应。 </li><li>回暖的0.05％胰蛋白酶EDTA，然后添加到覆盖细胞，板允许在室温下5分钟孵化。在这段时间内可以观察细胞四舍五入，和殖民地抬离板面。 </li><li>温和的反复吹打将删除从钢板表面的细胞和殖民地。 </li><li>放置在一个标准的锥形管，包含预热的胚胎干细胞媒体淬火剩下的胰蛋白酶，胰酶消化细胞，然后纺制。 </li><li>?...

<ol>
	<li>Cowan, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Derivation+of+Embryonic+Stem+Cells+from+Human+Blastocysts.">Derivation of Embryonic Stem Cells from Human Blastocysts.</a> , 1650-1656 (2004).</li></ol>

<p>冻结色调细胞株中描述的技术，我们有效地存储和复活的色调细胞株和利益的任何哺乳动物细胞。工作时，与胚胎干细胞系，染色体核型异常，可能会出现不正常的细胞可以迅速接管该行，冷冻的细胞在每一个通道的一小部分是一个重要的考虑因素。因此，有冷冻分割在某些段落中除了大银行，在每一个通道是非常可取的。 </p>

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>HuES Medium</td>
<td>medium</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>For aq total of 651.5 ml, add: 500 ml KO-DMEM + 6.5 ml PenStrep + 6.5 ml L- Glut + 6.5 ml NEAA 6.5 ml + beta-mercaptoethanol 0.6ul (14.3 M stock) + 65 ml Serum Replacement + 65 ml Plasmanate +  0.65 ml bFGF2 (~10 ng/mL final)
</td>
</tr>
<tr>
<td>Freezing Medium</td>
<td>medium</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>80% FCS (fetal calf serum) + 20% DMSO (dimethyl sulphoxide). Stored at 4C. </td>
</tr>		</tbody>
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freezing human es cells

<p class="jove_content">在这里，我们证明了我们的实验室如何冻结色调人类胚胎干细胞系。一个健康，成倍扩大培养，用PBS洗，以去除残留的媒体，否则可能淬火胰蛋白酶反应。回暖的0.05％胰蛋白酶EDTA，然后添加到覆盖细胞，板允许在室温下5分钟孵化。在这段时间内可以观察细胞四舍五入，和殖民地抬离板面。温和的反复吹打将删除从钢板表面的细胞和殖民地。放置在一个标准的锥形管，包含预热的胚胎干细胞媒体淬火剩下的胰蛋白酶，胰酶消化细胞，然后纺制。细胞悬浮在大约一百万个细胞的浓度生长介质，在一个媒体毫升，浓度等，一个冻结等分足够的复活一种文化上10厘米板。充分重悬细胞后，冰冷的冻结媒体是在同等体积增加。细胞悬浮液充分混合，分装成冷冻小瓶，允许慢慢冻结超过24小时- 80C。冷冻细胞可以长期贮存的液氮转移到气相，或约6个月保持在-80。</p>

Watch this Scientific Journal Video about Freezing Human ES Cells at JoVE.com

Freezing Human ES Cells

<p class="jove_content">在这里，我们证明了我们的实验室如何冻结色调人类胚胎干细胞系。</p>

冻结人类胚胎干细胞

Here we demonstrate how our lab freezes HuES human embryonic stem cell lines.  A healthy, exponentially expanding culture is washed with PBS to remove ...

Research

JoVE Journal

Biology

Human ES cells: Starting Culture from Frozen Cells

人类胚胎干细胞：启动文化冷冻细胞

Here we demonstrate how our lab begins a HuES human embryonic stem cell line culture from a frozen stock.  First, a one to two day old ten cm plate of  ...

科研

生物学

ES Cell-derived Neuroepithelial Cell Cultures

ES细胞源性神经上皮细胞培养

ES cells have the potential to differentiate into cells from all germ layers, which makes them an attractive tool for the development of new therapies. In ...

Propagation of Human Embryonic Stem (ES) Cells

Propagation of Human Embryonic Stem ES Cells

人类胚胎干细胞（ES细胞）的繁殖

Flash Freezing and Cryosectioning E12.5 Mouse Brain

闪存冻结和Cryosectioning E12.5小鼠脑

Freezing, Thawing, and Packaging Cells for Transport

冷冻，解冻，和运输包装细胞

Cultured mammalian cells are used extensively in cell biology studies. It requires a number of special skills in order to be able to preserve the ...

Freezing and Thawing Human Embryonic Stem Cells

人类胚胎干细胞冷冻和解冻

Since James Thomson et al developed a technique in 1998 to isolate and grow hES in culture, freezing cells for later use and thawing and expanding cells ...

Differentiation of Embryonic Stem Cells into Oligodendrocyte Precursors

胚胎干细胞分化成少突胶质前体

Oligodendrocytes are the myelinating cells of the central nervous system. For regenerative cell therapy in demyelinating diseases, there is significant ...

Modified ES / OP9 Co-Culture Protocol Provides Enhanced Characterization of Hematopoietic Progeny

修改ES / OP9共培养协议，提供增强造血后代的表征

The in vitro differentiation of ES cells towards a hematopoietic cell fate is useful when studying cell populations that are difficult to access in vivo ...

Transduction of Human Cells with Polymer-complexed Ecotropic Lentivirus for Enhanced Biosafety

增强生物安全与聚合物复合物Ecotropic慢病毒的人体细胞的转导

Stem and tumor cell biology studies often require viral transduction of human cells with known or suspected oncogenes, raising major safety issues for ...

Examination of <em>Drosophila</em> Larval Tracheal Terminal Cells by Light Microscopy

Examination of Drosophila Larval Tracheal Terminal Cells by Light Microscopy

考试<em&gt;果蝇</em幼虫气管终端细胞光镜

Cell shape is critical for cell function. However, despite the importance of cell morphology, little is known about how individual cells generate specific ...