Name: a standardized method for measuring internal lung surface area via mouse pneumonectomy and prosthesis implantation
Uploaded: 2017-07-26T15:00:00
Description: Watch this Scientific Journal Video about A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation at JoVE.com

作者要向北京国家生物科学研究所致谢。这项工作得到了北京市自然科学基金（Z17110200040000）的支持。

本议定书中使用的所有程序均按照北京国家生物科学研究所“实验动物护理和使用指南”中的建议进行。将8周龄的CD-1雄性小鼠饲养在特定的无病原体（SPF）设施中，直至进行实验。使用完全麻醉的小鼠进行手术（ 即 ，没有任何趾夹紧反应）。手术后，将小鼠保持在温暖，潮湿的房间内，食物和淡水充足。使用过量腹膜内注射的麻醉剂处死小鼠。 小鼠PNX手术<ol><li>通...

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N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Airspace+configuration+at+different+transpulmonary+pressures+in+normal+and+paraquat-induced+lung+injury+in+rats.">Airspace configuration at different transpulmonary pressures in normal and paraquat-induced lung injury in rats.</a> Am J Respir Crit Care Med. 158 (4), 1230-1234 (1998).</li><li>Yilmaz, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+quantification+of+heterogeneous+lung+growth+following+extensive+lung+resection+by+high-resolution+computed+tomography.">Noninvasive quantification of heterogeneous lung growth following extensive lung resection by high-resolution computed tomography.</a> J Appl Physiol. 107 (5), 1569-1578 (2009).</li><li>Voswinckel, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterisation+of+post-pneumonectomy+lung+growth+in+adult+mice.">Characterisation of post-pneumonectomy lung growth in adult mice.</a> Eur Respir J. 24 (4), 524-532 (2004).</li><li>Ravikumar, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regional+Lung+Growth+and+Repair+Regional+lung+growth+following+pneumonectomy+assessed+by+computed+tomography.">Regional Lung Growth and Repair Regional lung growth following pneumonectomy assessed by computed tomography.</a> J Appl Physiol. 97, 1567-1574 (2004).</li><li>Gibney, B. 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在本协议中，我们提供关于肺小鼠左肺PNX和假体植入后肺参数测量的详细描述。 ISA现在被认为是评估许多肺部疾病和损伤引起的肺泡再生中呼吸功能的关键指标。然而，虽然肺部研究界对ISA的实用性有一致意见，但迄今为止，几乎没有考虑用于计算ISA的两个参数ILV和MLI的测量标准化。显然，与任何测量一样，重要的是尝试获得无偏见的数据。本研究工作的核心目标是建立一个标准化的方案供小鼠...

肺的基本功能是血管和大气之间的氧气和二氧化碳的交换。肺部疾病，如支气管肺发育不良（BPD），慢性阻塞性肺病（COPD）和急性呼吸道感染，导致降低的ISA 2。研究人员在研究肺病已经开发出多种定量方法来评估肺形态变化，包括美林，ILV，3的气体交换单元的数量，ISA，和肺组织顺应性2。 Weibel 等人的开创性研究4和Duguid 等人 5一起确定了ISA可以作为人肺肺气交换能力的直接指标，可作为确定肺气肿严重程度的标准。过去五年发表的一些研究已经使用肺形态学参数（ 例如， </e米&gt; ISA和MLI）对发展6期间和从损伤PNX 1,7恢复期间评估在小鼠的肺中的形态和功能的变化。 ISA使用等式1 8,9计算： <img alt="方程" src="/files/ftp_upload/56114/56114eq1.jpg" /> ，其中ILV是内部肺容积，MLI是表示肺外周空间大小10的中间参数。 PNX，手术切除的一个或多个肺叶，已被广泛报道，以诱导在许多物种中肺泡再生，包括人类11，小鼠1，狗12，大鼠13，和兔14,15。一个螺柱PNX后14天的小鼠肺显示，既存在肺泡的扩张和肺泡从头形成有助于恢复其余肺组织中的ISA，ILV和肺泡数量1 。我们和其他人已经表明，在PNX（ 即 ，假体植入）后，将诸如海绵，蜡或定制假体的材料插入空胸腔会损害肺泡再生。现在已牢固地确立机械力功能的发起肺泡再生1，16，17最重要的因素之一。这些研究强调了使用PNX处理和假体植入肺的ISA值作为定量评估肺泡再生的标准的有效性。 已知观察者偏倚显着影响测量值肺部形态学参数（ 例如 ，MIL和ILV）。可以使用标准化协议来消除在确定ILV和MLI中的这种偏差，这是在ISA的计算中使用的两个参数。在这里，我们提供了高度详细的标准化方案来测量这些肺参数。重要的是，准确量化ISA的能力有望提高损伤诱导的肺泡再生模型中肺功能研究的可靠性和可重复性，并且有助于多种肺部疾病的机械发现。

<table width="881">
	<tbody>
		<tr height="21">
			<td height="21" style="width:411px;height:21px;">Low cost cautery kit</td>
			<td style="width:167px;">Fine Science Tools</td>
			<td style="width:136px;">18010-00</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Noyes scissors</td>
			<td>Fine Science Tools</td>
			<td>15012-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Standard pattern forceps</td>
			<td>Fine Science Tools</td>
			<td>11000-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Castroviejo Micro Needle Holders</td>
			<td>Fine Science Tools</td>
			<td>12060-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vessel clips</td>
			<td>Fine Science Tools</td>
			<td>18374-44</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">I. V. Cannula-20 gauge</td>
			<td>Jinhuan Medical Product Co., LTD.</td>
			<td>29P0601</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Surgical suture</td>
			<td>Jinhuan Medical Product Co., LTD.</td>
			<td>F602</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mouse intubation platform</td>
			<td>Penn-Century, Inc</td>
			<td>Model MIP</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Small Animal Laryngoscope</td>
			<td>Penn-Century, Inc</td>
			<td>Model LS-2-M</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TOPO Small Animal Ventilator</td>
			<td>Kent Scientific</td>
			<td>RSP1006-05L</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Thermal pad</td>
			<td>Stuart equipment</td>
			<td>SBH130D</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pentobarbital sodium salt</td>
			<td>Sigma</td>
			<td>P3761</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Heparin sodium salt</td>
			<td>Sigma</td>
			<td>H3393</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hematoxylin Solution</td>
			<td>Sigma</td>
			<td>GHS132</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Eosin Y solution, alcoholic</td>
			<td>Sigma</td>
			<td>HT110116</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 ml Pipette</td>
			<td>Thermo Scientific</td>
			<td>170356</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paraformaldehyde</td>
			<td>Sigma</td>
			<td>P6148</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">O.C.T Compound</td>
			<td>Tissue-Tek</td>
			<td>4583</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">cryosection machine</td>
			<td>Leica</td>
			<td>CM1950</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Disposable Base Molds</td>
			<td>Fisher HealthCare</td>
			<td>22-363-553</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">18 gauge needle</td>
			<td>Becton Dickinson</td>
			<td>305199</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Povidone iodine</td>
			<td>Fisher Scientific</td>
			<td>19-027132</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">70% ethanol</td>
			<td>Fisher Scientific</td>
			<td>BP82011</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Infusion sets for single use</td>
			<td>Weigao</td>
			<td>SFDA 2012 3661704</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Phosphate buffered saline</td>
			<td>Gibco</td>
			<td align="right">10010023</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tapes</td>
			<td>3M Scotch</td>
			<td style="width:136px;">8915</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cotton pad</td>
			<td>Vinda</td>
			<td>Dr.P</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Silicone prosthesis</td>
			<td>Custom made</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Brightfield microscope</td>
			<td>Olympus</td>
			<td>VS120</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ruler tool</td>
			<td>Adobe Photoshop</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

a standardized method for measuring internal lung surface area via mouse pneumonectomy and prosthesis implantation

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), representing the gas-exchange capacity of the lung, is a critical criterion to assess respiratory function. However, observer bias can significantly influence measured values for lung morphological parameters. The protocol that we describe here minimizes variations during measurements of two morphological parameters used for ISA calculation: internal lung volume (ILV) and mean linear intercept (MLI). Using ISA as a morphometric and functional parameter to determine the outcome of alveolar regeneration in both pneumonectomy (PNX) and prosthesis implantation mouse models, we found that the increased ISA following PNX treatment was significantly blocked by implantation of a prosthesis into the thoracic cavity1. The ability to accurately quantify ISA is not only expected to improve the reliability and reproducibility of lung function studies in injured-induced alveolar regeneration models, but also to promote mechanistic discoveries of multiple pulmonary diseases.

<html>我们在这里进行了PNX治疗组和假体植入（假体植入）组的实验。这些分组与我们研究组14之前发表的研究中使用的分组相同。 小鼠PNX和假体植入程序如图2所示。 8周龄的CD-1雄性小鼠用于手术和量化。在PNX治疗组和假体植入组中，左肺叶切除（ 图2A -</stron...

Watch this Scientific Journal Video about A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation at JoVE.com

A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation

内肺表面积（ISA）是评估肺部疾病和损伤诱导肺泡再生的肺形态和生理学的关键标准。我们在这里描述了一种标准化方法，可以最大限度地减少肺肺切除术和假体植入小鼠模型中ISA的测量偏差。

通过小鼠肺切除术和假体植入测量内部肺表面积的标准方法

Pulmonary morphology, physiology, and respiratory functions change in both physiological and pathological conditions. Internal lung surface area (ISA), ...

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Production and Administration of Therapeutic Mesenchymal Stem/Stromal Cell (MSC) Spheroids Primed in 3-D Cultures Under Xeno-free Conditions

Production and Administration of Therapeutic Mesenchymal Stem/Stromal Cell MSC Spheroids Primed in 3-D Cultures Under Xeno-free Conditions

生产和治疗间充质干行政/基质细胞（MSC）球体底漆在三维培养下无异物条件

Mesenchymal stem/stromal cells (MSCs) hold great promise in bioengineering and regenerative medicine. MSCs can be isolated from multiple adult tissues via ...

Isolation and Enrichment of Liver Progenitor Subsets Identified by a Novel Surface Marker Combination

分离和新型表面标记组合来标识肝祖亚群富集

During chronic liver injuries, progenitor cells expand in a process called ductular reaction, which also entails the appearance of inflammatory cellular ...

Assessing Cardiomyocyte Subtypes Following Transcription Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts

评估心肌亚型以下鼠标胚胎成纤维细胞的转录因子介导的重编程

Direct reprogramming of one cell type into another has recently emerged as a powerful paradigm for regenerative medicine, disease modeling, and lineage ...

A Standardized Approach for Multispecies Purification of Mammalian Male Germ Cells by Mechanical Tissue Dissociation and Flow Cytometry

通过机械组织分离和流式细胞术对哺乳动物雄性生殖细胞多特异性纯化的标准化方法

Fluorescence-activated cell sorting (FACS) has been one of the methods of choice to isolate enriched populations of mammalian testicular germ cells. ...

A Method for Characterizing Embryogenesis in Arabidopsis

A Method for Characterizing Embryogenesis in Arabidopsis

Given the highly predictable nature of their development, Arabidopsis embryos have been used as a model for studies of morphogenesis in plants. However, ...

A Novel Use of Three-dimensional High-frequency Ultrasonography for Early Pregnancy Characterization in the Mouse

三维高频超声对小鼠早期妊娠特征的新应用

High-frequency ultrasonography (HFUS) is a common method to non-invasively monitor the real-time development of the human fetus in utero. The mouse is ...

Application of Chronic Stimulation to Study Contractile Activity-induced Rat Skeletal Muscle Phenotypic Adaptations

慢性刺激在收缩活动性大鼠骨骼肌表型适应性研究中的应用

Skeletal muscle is a highly adaptable tissue, as its biochemical and physiological properties are greatly altered in response to chronic exercise. To ...

Partial Lobular Hepatectomy: A Surgical Model for Morphologic Liver Regeneration

部分小叶切除术: 形态学肝再生的外科模型

Morphological organ regeneration following acute tissue loss is common among lower vertebrates, but is rarely observed in mammalian postnatal life. Adult ...