Name: primary human bronchial epithelial cells grown from explants
Uploaded: 2010-03-26T17:10:00
Description: Watch this Scientific Journal Video about Primary Human Bronchial Epithelial Cells Grown from Explants at JoVE.com

作者非常感谢理查德Inculet博士提供的支气管组织。获得圣若瑟医疗汉密尔顿和西安大略/伦敦健康科学中心（博士大卫马克），组织和档案馆委员会，病理科大学研究伦理委员会的批准。我们也感谢提供一些对免疫所需的材料提供电信设备制造商，我们的ALI的文化和丹妮拉法卡斯厄尼 - 斯皮策（电子显微镜，加拿大麦克马斯特大学）。这项工作是由一个从加拿大安大略省胸科协会座长期授予;阿斯玛亚吉博士FSORC奖学金，圣若瑟医疗保健，加拿大安大略省汉密尔顿，的支持。

人支气管段原发性支气管上皮细胞提供了一个丰富的源泉。在这篇文章中，我们描述一个人支气管上皮细胞（HBE）的新鲜分离的人支气管段的增长和扩张的协议。该协议包括五个部分： <ol><li>涂料和刮伤100毫米培养板</li><li>准备支气管组织植</li><li>支气管组织移植</li><li> HBE细胞传代</li><li> Transwells纤毛HBE细胞成长</li></ol> 开始之前 ，所有步骤都在生物安全柜（BSC?...

<ol>
	<li>Phillips, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growing+Cells+on+Transwell+Inserts+-+Tips+and+Techniques+[Internet].">Growing Cells on Transwell Inserts - Tips and Techniques [Internet].</a> Corning Life Sciences, Technical Resources, Online Training. , (2008).</li><li>Turi, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxidative+stress+activates+anion+exchange+protein+2+and+AP-1+in+airway+epithelial+cells.">Oxidative stress activates anion exchange protein 2 and AP-1 in airway epithelial cells.</a> Am J Physiol Lung Cell Mol. Physiol. 283, L791-L798 (2002).</li><li>Lechner, J. F., Haugen, A., McClendon, I. A., Pettis, E. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clonal+growth+of+normal+adult+human+bronchial+epithelial+cells+in+a+serum-free+medium.">Clonal growth of normal adult human bronchial epithelial cells in a serum-free medium.</a> In Vitro. 18, 633-642 (1982).</li><li>Yoshisue, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+ciliated+bronchial+epithelium+1,+a+ciliated+cell-associated+gene+induced+during+mucociliary+differentiation.">Characterization of ciliated bronchial epithelium 1, a ciliated cell-associated gene induced during mucociliary differentiation.</a> Am. J. Respir. Cell Mol. Biol. 31, 491-500 (2004).</li><li>Wetering, S. v. a. n. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+secretory+leukocyte+proteinase+inhibitor+(SLPI)+production+by+human+bronchial+epithelial+cells:+increase+of+cell-associated+SLPI+by+neutrophil+elastase.">Regulation of secretory leukocyte proteinase inhibitor (SLPI) production by human bronchial epithelial cells: increase of cell-associated SLPI by neutrophil elastase.</a> J. Investig. Med. 48, 359-366 (2000).</li><li>Tristram, D. A., Hicks, W., Hard, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Respiratory+syncytial+virus+and+human+bronchial+epithelium.">Respiratory syncytial virus and human bronchial epithelium.</a> Arch. Otolaryngol. Head Neck Surg. 124, 777-783 (1998).</li><li>Mattinger, C., Nyugen, T., Schafer, D., Hormann, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+serum-free+culture+conditions+for+primary+human+nasal+epithelial+cells.">Evaluation of serum-free culture conditions for primary human nasal epithelial cells.</a> Int. J. Hyg. Environ. Health. 205, 235-238 (2002).</li><li>Freshney, R. I., Freshney, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Culture of epithelial cells. , 1-23 (1992).</li><li>Freshney, R. I., Freshney, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Normal+human+bronchial+epithelial+cell+cultures+in+Culture+of+specialized+cells+series.">Normal human bronchial epithelial cell cultures in Culture of specialized cells series.</a> Culture of epithelial cells. , 181-196 (1992).</li></ol>

在这项研究中，我们提出了详细的文化和扩大初级人支气管上皮细胞的方法。我们演示了如何植和支气管组织，促进上皮细胞生长的媒体培养，移植可以提供连续无差异（淹没）和差异（气 - 液界面上的）的细胞模型研究了人类呼吸道上皮细胞源。在药物检测系统，更接近其真正的生理环境中的细胞，这些细胞可以利用。这是非常重要的，你看从植/移植的细胞生长，并确认的鹅卵石形态。如果细...

其它要求：孵化器，生物安全柜（BSC），离心机，100毫米的培养皿，无菌试管（15毫升，50毫升和2毫升），无菌枪头，手术刀把手和刀片，小锋利的剪刀，实验室外套和手套。这些可从您的首选供应商。

primary human bronchial epithelial cells grown from explants

人支气管上皮细胞需要为疾病的细胞模型，并探讨人类上皮细胞的功能和结构上的辅料和药物制剂的效果。在这里，我们详细描述了支气管气道组织是由外科医生在肺部手术（如肺癌或肺减容手术）的次收获的支气管上皮细胞生长的方法。伦理委员会批准和知情同意，医生拿了什么是需要病理，并为我们提供了一个支气管部分，是从病变区域的远程。该组织是作为一个日益增​​长的文化原发性支气管上皮细胞，可用于使用的植源。支气管段约0.5 - 1cm长和≤直径1cm冲洗用冷水EBSS和除去多余的实质组织。段切开，成2 - 3mm的肉末<sup&gt; 3</sup&gt;组织块。这些作品是用来作为原代细胞源。经过1-2小时100毫米文化涂层板的胶原蛋白相结合（30微克/毫升），纤维连接蛋白（10微克/毫升），与牛血清白蛋白（10微克/毫升），在4-5个地区和组织的钢板被划伤件放置在划痕地区，然后加入培养基（DMEM / HAM用添加剂的F - 12）适用于上皮细胞的生长和板被放置在一个孵化器在37 ° C，5％CO<sub&gt; 2</sub&gt;加湿空气。改变培养基每隔3-4天。上皮细胞的生长，形成约1.5厘米，直径3-4周环件。植，可重新使用多达6次转移到新的预涂层钢板。细胞被取消，使用胰蛋白酶/ EDTA，汇总，计算，在T75细胞绑定烧瓶中重新镀金，以增加它们的数量。 T75烧瓶种子2-3万细胞生长在4个星期到80％汇合。扩大初级人类的上皮细胞是可以培养的气 - 液界面上区分。这里描述的方法提供了丰富的新鲜分离的组织的人支气管上皮细胞源，并允许作为疾病模型，为研究这些细胞和药理学和毒理学筛选。

Watch this Scientific Journal Video about Primary Human Bronchial Epithelial Cells Grown from Explants at JoVE.com

Primary Human Bronchial Epithelial Cells Grown from Explants

在这里，我们描述一个人支气管气道组织的外植体，包括气 - 液界面上的区别的增长为发展初级人支气管上皮细胞的具体方法。这种方法提供了一个丰富的原代细胞源调查在人的肺部健康和疾病的气道上皮细胞的作用。

从外植体生长的首要人权支气管上皮细胞

Human bronchial epithelial cells are needed for cell models of disease and to investigate the effect of excipients and pharmacologic agents on the ...

Research

JoVE Journal

Biology

Preparation and Culture of Rat Lens Epithelial Explants for Studying Terminal Differentiation

大鼠晶状体上皮细胞外植体的制备及文化学习终末分化

The anterior surface of the ocular lens is covered by a monolayer of epithelial cells, which proliferate in an annular zone underlying the ciliary body.  ...

科研

生物学

Isolation and Culture of Adult Epithelial Stem Cells from Human Skin

成人从人体皮肤的上皮干细胞的分离和培养

The homeostasis of all self-renewing tissues is dependent on adult stem cells. As undifferentiated stem cells undergo asymmetric divisions, they generate ...

Ex Vivo Culture of Primary Human Fallopian Tube Epithelial Cells

Ex Vivo Culture of Primary Human Fallopian Tube Epithelial Cells

前初级人类输卵管上皮细胞的体外培养

Epithelial ovarian cancer is a leading cause of female cancer mortality in the United States. In contrast to other women-specific cancers, like breast and ...

Na&iuml;ve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

NaÃƒÆ’Ã‚Â¯ve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

天真成人干细胞的分离从初级人类成纤维细胞培养

Over the last decade, several adult stem cell  populations have been identified in human skin 1-4.  The isolation of multipotent adult dermal precursors ...

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 ...

Optimal Lentivirus Production and Cell Culture Conditions Necessary to Successfully Transduce Primary Human Bronchial Epithelial Cells

优化慢病毒生产和细胞培养必要条件，成功转导初级人支气管上皮细胞

In vitro culture of primary human bronchial epithelial (HBE) cells using air-liquid interface conditions provides a useful model to study the processes of ...

Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D

方法来研究上皮转运蛋白功能和表达在纯肠和Caco-2细胞生长在3D

The intestinal epithelium has important transport and barrier functions that play key roles in normal physiological functions of the body while providing ...

Multicellular Human Alveolar Model Composed of Epithelial Cells and Primary Immune Cells for Hazard Assessment

由上皮细胞和原发免疫细胞组成的多细胞人类Alveal模型，用于危险评估

A human alveolar cell coculture model is described here for simulation of the alveolar epithelial tissue barrier composed of alveolar epithelial type II ...

Generation of Murine Primary Colon Epithelial Monolayers from Intestinal Crypts

来自肠道密码的穆林原结肠上皮单层的生成

The intestinal epithelium is comprised of a single layer of cells that act as a barrier between the gut lumen and the interior of the body. Disruption in ...

Efficient Dissection and Culture of Primary Mouse Retinal Pigment Epithelial Cells

原发性小鼠视网膜色素上皮细胞的有效解剖与培养

Eye disorders affect millions of people worldwide, but the limited availability of human tissues hinders their study. Mouse models are powerful tools to ...