Name: isolation and animal serum free expansion of human umbilical cord derived mesenchymal stromal cells (mscs) and endothelial colony forming progenitor cells (ecfcs)
Uploaded: 2009-10-08T16:03:00
Description: Watch this Scientific Journal Video about Isolation and Animal Serum Free Expansion of Human Umbilical Cord Derived Mesenchymal Stromal Cells MSCs and Endothelial Colony Forming Progenitor Cells ECFCs

作者感谢提供有用的意见，手稿，丹妮拉泰勒和Margaretha神父hwirth提供技术援助，莫妮卡语言编辑法雷尔，乌韦郎，马尔吉特Holzapfel和桑德拉部Eppich妇产科pHPL，伊娃罗德和妮可霍夫曼卡塔琳娜Schallmoser提供的线。 这项工作一直由奥地利研究基金会（FWF，授予N211南到DS），奥地利研究促进署（FFG，授予N200到DS）和成体干细胞研究基金会（AR）的支持。 AR是博士课程的格拉茨医科大学分子医学研究员。

第1部分：设置 1。细胞培养液的制备在开始培养基的配制之前，收集所有的材料和工具，你需要。 解冻2 × 56毫升分装汇集了人类的血小板裂解（pHPL，自制：参考文献1的Jove＃1523），一个100X青霉素/链霉素溶液，10毫升× 5毫升（L -谷氨酰胺两个西格玛）。 解冻的细胞因子和生长因子等分（血管内皮生长因子，碱性成纤维细胞生长因子...

<ol>
	<li>Schallmoser, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+platelet+lysate+can+replace+fetal+bovine+serum+for+clinical-scale+expansion+of+functional+mesenchymal+stromal+cells.">Human platelet lysate can replace fetal bovine serum for clinical-scale expansion of functional mesenchymal stromal cells.</a> Transfusion. 47, 1436-1446 (2007).</li><li>Reinisch, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+system+to+propagate+cord+blood-derived+multipotent+mesenchymal+stromal+cells+for+clinical+application.">Humanized system to propagate cord blood-derived multipotent mesenchymal stromal cells for clinical application.</a> Regen Med. 2, 371-382 (2007).</li><li>Reinisch, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+large-scale+expanded+endothelial+colony-forming+cells+function+in+vitro+and+in+vivo.">Humanized large-scale expanded endothelial colony-forming cells function in vitro and in vivo.</a> Blood. 113, 6716-6725 (2009).</li><li>Strunk, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenotypic+characterization+and+preclinical+production+of+human+lineage-negative+cells+for+regenerative+stem+cell+therapy.">Phenotypic characterization and preclinical production of human lineage-negative cells for regenerative stem cell therapy.</a> Transfusion. 45, 315-326 (2005).</li></ol>

有一个从中获取干细胞或ECFCs包括骨髓，脂肪组织，脐带血，等各种来源从相同的源祖细胞两种类型直接比较的样组织和血管再生或肿瘤进展的贡献过程中的不同类型的细胞参与，这是必要的。 隔离，并随后从同一供体研究ECFCs和干细胞自体对缓解使得这种方法排除在个别实验的捐助变化的优势。 与造血干细胞可能造成的污染降到最?...

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<td>alpha modified MEM</td>
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<td>L-GLutamin</td>
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<td>Penicillin/Streptomycin</td>
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<td>Sigma</td>
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<td>EBM</td>
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<td>Lonza</td>
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<td>Single cytokine quots</td>
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<td>Lonza</td>
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<td>75 cm2 flasks</td>
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<td>Corning</td>
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<td>150 cm2 plates</td>
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<td>Cell scraper</td>
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<td>Corning</td>
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<td>Trypsin/EDTA</td>
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<td>Sigma</td>
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<td>Monoclonal antibodies</td>
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<td>Becton Dickenson</td>
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<td>PBS</td>
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<td>50 ml tubes</td>
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<td>Falcon</td>
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<td>500 mL filter 20 &mu;m pore size </td>
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<td>Millipore</td>
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<td>Preservative free heparin</td>
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isolation and animal serum free expansion of human umbilical cord derived mesenchymal stromal cells (mscs) and endothelial colony forming progenitor cells (ecfcs)

脐带是一种具有高增殖潜能，包括间质的基质干细胞（也称为间质干细胞，MSCS）和内皮细胞集落形成祖细胞（ECFCs）的祖细胞的丰富来源。这两种类型的细胞，在维持组织的完整性关键人物，并可能也参与再生过程和肿瘤的形成。 要比较的方式研究它们的生物学和功能，它是重要的，都从同一供体细胞类型。这也可能是来自同一组织的干细胞和ECFCs，有利于再生的目的。 由于细胞疗法，最终找到自己的方式从替补到床边，我们建立了一个新的方法来隔离和进一步扩大不使用动物性蛋白质的祖细胞。在我们的协议中的所有步骤汇集人类血小板裂解液（pHPL）所取代胎牛血清完全避免接触细胞异种蛋白质。 该视频演示了从一个脐带祖细胞的分离和扩张的方法。 所有材料和程序进行说明。

Watch this Scientific Journal Video about Isolation and Animal Serum Free Expansion of Human Umbilical Cord Derived Mesenchymal Stromal Cells MSCs and Endothelial Colony Forming Progenitor Cells ECFCs

Isolation and Animal Serum Free Expansion of Human Umbilical Cord Derived Mesenchymal Stromal Cells MSCs and Endothelial Colony Forming Progenitor Cells ECFCs

这个协议描述不使用动物血清中产生自体对实验移植的目的的间质基质细胞和内皮细胞集落形成细胞的分离和随后的扩大。

隔离和动物血清自由膨胀的人脐血源性间质基质细胞（MSCs），内皮细胞集落形成祖细胞（ECFCs）

The umbilical cord is a rich source for progenitor cells with high proliferative potential including mesenchymal stromal cells (also termed mesenchymal ...

Research

JoVE Journal

Biology

Isolation and Animal Serum Free Expansion of Human Umbilical Cord Derived Mesenchymal Stromal Cells (MSCs) and Endothelial Colony Forming Progenitor Cells (ECFCs)

Isolation of Human Umbilical Vein Endothelial Cells (HUVEC)

Isolation of Human Umbilical Vein Endothelial Cells HUVEC

分离人脐静脉内皮细胞（HUVEC）

Angiogenesis is a complex multi-step process, where in response to angiogenic stimuli, new vessels are created from the existing vasculature. These steps ...

科研

生物学

Isolation and Large Scale Expansion of Adult Human Endothelial Colony Forming Progenitor Cells

成人内皮细胞集落形成祖细胞的分离和大型规模扩张

This paper introduces a novel recovery strategy for endothelial colony forming progenitor cells (ECFCs) from heparinized but otherwise unmanipulated adult ...

Isolation and Enrichment of Rat Mesenchymal Stem Cells (MSCs) and Separation of Single-colony Derived MSCs

Isolation and Enrichment of Rat Mesenchymal Stem Cells MSCs and Separation of Single-colony Derived MSCs

分离和富集大鼠间质干细胞（MSCS）及衍生的干细胞分离单细胞集落

MSCs are a population of adult stem cells that is a promising source for therapeutic applications. These cells can be isolated from the bone marrow and ...

Isolation of Human Umbilical Arterial Smooth Muscle Cells (HUASMC)

Isolation of Human Umbilical Arterial Smooth Muscle Cells HUASMC

人脐动脉平滑肌细胞的分离（HUASMC）

The human umbilical cord (UC) is a biological sample that can be easily obtained just after birth. This biological sample is, most of the time, discarded ...

Feeder-Free Adaptation, Culture and Passaging of Human IPS Cells using Complete KnockOut Serum Replacement Feeder-Free Medium

免费接驳适应，文化和传代使用完整的基因敲除血清替代馈线中的人类iPS细胞

The discovery in 2006 that human and mouse fibroblasts could be reprogrammed to generate iPS cells 1-3 with qualities remarkably similar to embryonic stem ...

Phenotypic and Functional Characterization of Endothelial Colony Forming Cells Derived from Human Umbilical Cord Blood

来自人类脐带血内皮细胞集落形成细胞的表型和功能特性

Longstanding views of new blood vessel formation via angiogenesis, vasculogenesis, and arteriogenesis have been recently reviewed1. The presence of ...

Isolation and Differentiation of Stromal Vascular Cells to Beige/Brite Cells

米色/布里特细胞的间质血管细胞的分离和分化

Brown adipocytes have the ability to uncouple the  respiratory chain in mitochondria and dissipate chemical energy as heat.  Development of UCP1-positive ...

Induction and Analysis of Epithelial to Mesenchymal Transition

诱导上皮和分析间质转化

Epithelial  to mesenchymal transition (EMT) is essential for proper morphogenesis during  development. Misregulation of this  process has been implicated ...

Isolation of Murine Valve Endothelial Cells

鼠阀门内皮细胞的分离

Normal valve structures consist of stratified layers of specialized extracellular matrix (ECM) interspersed with valve interstitial cells (VICs) and ...

Isolation of CD146+ Resident Lung Mesenchymal Stromal Cells from Rat Lungs

Isolation of CD146+ Resident Lung Mesenchymal Stromal Cells from Rat Lungs

CD146的分离<sup&gt; +</sup&gt;从大鼠肺组织驻地肺间质基质细胞

Mesenchymal stromal cells (MSCs) are increasingly recognized for their therapeutic potential in a wide range of diseases, including lung diseases. Besides ...