这项工作由巴黎狄德罗大学、法国国立卫生和医学研究研究所 (INSERM) 和法国牙科医生研究所 (IFRO) 资助。

在本研究中使用的所有动物都是按照法国农业部 (A-75-06-12) 的护理和使用实验动物的指导方针维持的。
1. 动物接触毒物
<ol><li>在执行本议定书之前, 获得必要的机构批准, 并确保遵守所有动物护理指南。</li>
	<li>应用研究协议的设计, 允许不同实验组的组成, 以测试分子对成釉细胞分泌期和成釉细胞成熟期的影响。在这里, 四组雄性大鼠的组成取决于他们接触氟化物 (NaF) 组合或不与 BPA (图 1A)2。所有动物在65天被观察和解剖 (图 1B)。</li>
</ol>2. 成年大鼠半下颌的解剖
<ol><li>弄死大鼠由 CO2窒息, 可选择地然后被斩首对分开的头从身体的其他。在专用框16中, 向2中的老鼠公开5分钟。</li>
	<li>使用 #11 手术刀去除下唇上的所有皮肤, 以便方便地接触下切牙。</li>
	<li>用同样的手术刀, 在两个较低的门牙之间进行切口轻微的压力, 下颌将被分成两半。</li>
	<li>用手术刀切开颞下颌关节, 将下颚分离, 用细镊子握住半下颌。</li>
	<li>用手术刀切开周围的软组织, 用刮刀除去所有的肌肉、肌腱和韧带, 直到骨头完全干净为止。</li>
</ol>3. 切牙的隔离17,18
<ol><li>将刀片平行于切牙的主要纵轴, 仔细地剃掉骨头。开始在骨脊附近的门牙尖端 (近端), 直到年底的门牙附近的颈部循环。切口运动从近端向远端方向进行。</li>
	<li>用手术刀将第一切口远端至颈环, 以除去半下颌的 gonion, 并允许在不损害宫颈环或成釉细胞分泌阶段的情况下轻易脱去门牙(图2中的红线)。</li>
	<li>在第二个磨牙下面做第二次切割, 并在切牙的骨和内侧表面插入手术刀。当所有的基底骨被解除, 旋转向外切牙, 并采取了小心, 以避免搪瓷器官组织损害使用细镊子。</li>
</ol>4. 双目镜下搪瓷器官的显微解剖
<ol><li>用吸管将200µL 盐磷酸盐缓冲液 (PBS 1X) 放在切牙上。用细镊子与唇面朝上。在组织的无色和橙色部分之间做手术刀标记。此标记对应于随后可观察到的底层白点 (图 2)。</li>
	<li>刮, 用挖掘机或等效工具, 细胞表面从手术刀标记到顶端末端对应于分泌阶段 (无色) 成釉细胞, 并且从手术刀标记到门牙尖端为成熟阶段 (橙色) 成釉细胞。</li>
	<li>移除与过渡阶段相对应的 2 mm 组织, 如前面描述的19所述。在搪瓷器官解剖时不要打开门牙, 以免被间质污染。</li>
	<li>将位于门牙顶端部分 (图 2) 的颈部环切开, 如前所述20所示。</li>
	<li>收集10% 福尔马林或溶解溶液进行进一步的调查 (请参阅材料表)。</li>
</ol>5. 收集分离的搪瓷器官组织进行进一步调查
<ol><li>将200µL PBS 1X 缓冲器放在切牙上。</li>
	<li>用 #11 手术刀刀片, 仔细地分离牙上皮细胞逐渐在缓冲, 分别为每个阶段在手术刀标记之前做的帮助。</li>
	<li>对于细胞 RNA 和蛋白质提取, 把组织的溶解溶液, 允许提取蛋白质和 rna (参见材料表)。 注: 通过转动试管中的组织挖掘机, 然后研磨细胞直至获得均匀的混合物, 制备高质量的 rna。该混合物已准备好 RNA 和蛋白质提取根据制造商的过程, 这是以前描述的2,8,17。通常, 每种制剂都能获得大约5-10 µg 总 rna 和150µg 总蛋白。</li>
	<li>对于组织学分析, 免疫组化 (IHC) 和原位杂交, 把细胞层在10% 福尔马林2小时, 然后存储在4摄氏度在 PBS 1X。组织可以嵌入蜡/石蜡或组织 OCT 的冰冻切片。</li>
	<li>对于 EMP 提取物, 用细镊子取门牙。在短时间暴露于空气 (轻微脱水) 后, 在切牙的唇表面的中间部分可以看到白色斑点, 这代表了搪瓷矿化的起始。这个白点对应于成釉细胞的过渡/早期成熟阶段, 所以使用它作为提取EMPs21 的指示器。</li>
</ol>

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作者没有披露的利益冲突。

改变成釉细胞活动和/或扰乱成釉细胞的扩散, 分化和成熟过程导致不可逆的搪瓷缺陷, 反过来, 搪瓷缺陷的表征可能有助于发展对改变的理解成釉细胞活动在 amelogenesis 期间。因此, 对分离的搪瓷器官的研究是决定其起源、环境或遗传的病理事件的决定性因素。
此技术最初是由希勒et描述的。17和罗宾逊et 。18, 用于显示对搪瓷器?...

许多发展中的搪瓷缺陷可能是由于暴露于环境毒物和/或不适当的生活方式1,2,3,4。使用目前所描述的程序来破坏 amelogenesis 事件和分子的特性, 将促进使用由此产生的搪瓷缺陷, 作为暴露于几种毒物的早期标记, 并有助于重建健康史。每个病人在围产期期间, 搪瓷是综合升华 1, 2.搪瓷合成可分为四个主要阶段, 取决于成釉细胞活动5。第一步重整旗鼓前体细胞和成釉细胞的增殖。在第二步, 分化成釉细胞分泌搪瓷基质蛋白 (EMPs), 主要釉原蛋白, 釉蛋白和 ameloblastin, 这决定了最终搪瓷的厚度。因此, 电磁脉冲合成的任何干扰都会导致搪瓷的数量缺陷。在充分的搪瓷厚度的沉积以后, 成熟阶段开始。在这个阶段, 磷灰石微晶生长的宽度和厚度, 使搪瓷达到最高的矿化比发现在一个生物组织, 多达96% 的重量。破坏在成熟阶段发生的事件导致质量搪瓷缺陷。最后, 成釉细胞进入成熟后的阶段, 也称为啮齿动物的色素沉着, 在牙齿喷发过程中发生凋亡, 使搪瓷缺损 (如有) 无法挽回和不可逆转, 因此缺陷提供了潜在的回顾性记录。成釉细胞强调。在啮齿类动物中, amelogenesis 遵循类似的事件序列, 其门牙的特殊性不断增加, 这使得它们成为研究 amelogenesis 一般过程的合适模型。因此, amelogenesis 的任何中断都会导致搪瓷质量和/或数量的改变, 这取决于中断事件的时间窗口。在这个意义上, 暴露于二恶英, 铅和内分泌干扰化学品 (EDCs), 如双酚 a (BPA), 染料木黄酮和 vinclozolin, 已被证明生成搪瓷 hypomineralizations1,2,3 ,6,7,8。在胎儿期和出生后的第一个月内, 在低剂量 BPA 剂量的大鼠的门牙上发现不对称的白色不透明斑点1。这些搪瓷缺陷的大鼠, 以及人磨牙切牙 hypomineralization (MIH), 具有相似的临床, 结构, 和生物化学的特点。MIH 是最近被描述的牙釉质病理学, 病因仍然是不明的9,10尽管许多原因因素被假设了9,10,11 ,12。
另一重要的搪瓷 hypomineralization 病理由于环境因素是牙氟中毒 (DF), 这是由于过量氟化物吸收 (> 0.1 毫克/千克/天)13,14。氟化物的主要来源是饮用的水, 补充或自然富含氟化物。氟化物也经常被规定防止龋齿, 但预防剂量仅比毒性的50% 低 (≤0.05 毫克/公斤/天)。MIH 和 DF, 由于暴露于环境因素而引起的两个常见的病症, 可能呈现出共同的特征, 因为氟化物与其他有毒物质 (如 EDCs2 ) 的 hypomineralizing 效应增强, 因此需要加以表征。或阿莫西林15。
在不同分化阶段对成釉细胞的大鼠釉质器官进行显微解剖, 有助于了解分子的作用机制, 能够扰乱成釉细胞活性, 导致牙釉质缺损在牙齿喷发后确诊。换言之, 由于环境毒物的影响, 搪瓷基因表达的变化和搪瓷基质成分的改变, 使暴露于毒物的历史得以重建, 并促进对公众的环境安全监测。健康.

<table>
	<tbody>
		<tr>
			<td>Bisphenol A</td>
			<td>Sigma Aldrich, Saint Louis MO</td>
			<td>239658</td>
			<td></td>
		</tr>
		<tr>
			<td>formalin 10%</td>
			<td>Sigma-Aldrich, Saint Louis, MO</td>
			<td>HT5012</td>
			<td></td>
		</tr>
		<tr>
			<td>Tri-Reagent</td>
			<td>Euromedex, France</td>
			<td>TR118</td>
			<td></td>
		</tr>
		<tr>
			<td>RLT buffer</td>
			<td>Qiagen, Les Ulis, France</td>
			<td>74126</td>
			<td>RNeasy Protect Mini Kit</td>
		</tr>
		<tr>
			<td>Androgen receptor antibody</td>
			<td>Santa Cruz Biotechnology, Santa Cruz, CA)</td>
			<td>sc-816</td>
			<td>rabbit polyclonal antibody</td>
		</tr>
		<tr>
			<td>PBS 10x</td>
			<td>EUOMEDEX</td>
			<td>ET330.A</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium fluoride (NaF)</td>
			<td>Sigma-Aldrich, Saint Louis, MO</td>
			<td>S-1504</td>
			<td></td>
		</tr>
		<tr>
			<td>paraplast regular</td>
			<td>Leica microsystems, Nanterre cedex,&#160;France</td>
			<td>39601006</td>
			<td>called was/parafin in the text</td>
		</tr>
		<tr>
			<td>tissue OCT</td>
			<td>VWR, Fontenay-sous-Bois, France</td>
			<td>411243</td>
			<td></td>
		</tr>
		<tr>
			<td>Extra Fine Bonn Scissors - Straight/8.5 cm</td>
			<td>PHYMEP , Paris, France</td>
			<td>14084-08</td>
			<td></td>
		</tr>
		<tr>
			<td>Handle for Scalpel Blades - 12.5 cm</td>
			<td>PHYMEP, Paris, France</td>
			<td>10035-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved Scalpel Blade</td>
			<td>PHYMEP , Paris, France</td>
			<td>10035-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting Knife - Fine/Straight Tip</td>
			<td>PHYMEP , Paris, France</td>
			<td>10055-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Circle Knife</td>
			<td>PHYMEP, Paris, France</td>
			<td>10059-15</td>
			<td></td>
		</tr>
		<tr>
			<td>scalpel blades n&#176;11 Swann-Morton</td>
			<td>VWR, Fontenay-sous-Bois, France</td>
			<td>233-0024</td>
			<td></td>
		</tr>
		<tr>
			<td>binocular lens</td>
			<td>Leica biosystems, Nanterre cedex,&#160;France</td>
			<td>MZFLIII</td>
			<td></td>
		</tr>
	</tbody>
</table>

micro-dissection of enamel organ from mandibular incisor of rats exposed to environmental toxicants

由于环境条件和生活方式造成的搪瓷缺陷是公共卫生关注的问题, 因为它们的发病率很高。这些缺陷是由负责搪瓷合成的细胞的活性改变所造成的, 成釉细胞是在搪瓷器官中存在的。在 amelogenesis 期间, 成釉细胞遵循一个特定的和精确的序列的事件的扩散, 分化和死亡。大鼠不断生长的门牙是研究成釉细胞活性和分化阶段的一种适宜的实验模型。在这里, 我们描述了一个可靠和一致的方法, 对暴露于环境毒物的大鼠的牙釉质器官进行显微解剖。微解剖的牙科上皮含有分泌物和成熟阶段的成釉细胞, 可用于定性实验, 如免疫组化分析和原位杂交, 以及定量分析, 如qPCR, RNA 序列, 和西方印迹。

许多搪瓷缺陷, 如牙科氟中毒12,13, 可能是由于环境条件由于过量氟化物吸收或搪瓷 hypomineralization 类似 MIH 由于暴露在一些 EDCs1, 7,22。这些发育的搪瓷缺陷可在大鼠身上进行实验复制 (图 1)1,2<s...

Watch this Scientific Journal Video about Micro-dissection of Enamel Organ from Mandibular Incisor of Rats Exposed to Environmental Toxicants at JoVE.com

Micro-dissection of Enamel Organ from Mandibular Incisor of Rats Exposed to Environmental Toxicants

了解搪瓷的形成和可能的改变需要研究成釉细胞活动。在这里, 我们描述了一个可靠和一致的方法, 显微解剖搪瓷器官含有分泌物和成熟阶段成釉细胞, 可用于进一步定量和定性实验程序。

环境毒物暴露大鼠下颌切牙釉质器官的显微解剖

Enamel defects resulting from environmental conditions and ways of life are public health concerns because of their high prevalence. These defects result ...

micro-dissection-enamel-organ-from-mandibular-incisor-rats-exposed-to

Research

JoVE Journal

Medicine

10.0K 次观看.  Cordeliers Research Centre. 了解搪瓷的形成和可能的改变需要研究成釉细胞活动。在这里, 我们描述了一个可靠和一致的方法, 显微解剖搪瓷器官含有分泌物和成熟阶段成釉细胞, 可用于进一步定量和定性实验程序。

视频: Micro-dissection of Enamel Organ from Mandibular Incisor of Rats Exposed to Environmental Toxicants

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

测量和制止在弱视治疗

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

科研

医学

Fundus Photography as a Convenient Tool to Study Microvascular Responses to Cardiovascular Disease Risk Factors in Epidemiological Studies

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an important role in maintaining cardiovascular health. The retina is a tissue that lines the interior of the eye and it is the only tissue that allows for a non-invasive analysis of the microvasculature. Nowadays, high-quality fundus images can be acquired using digital cameras. Retinal images can be collected in 5 min or less, even without dilatation of the pupils. This unobtrusive and fast procedure for visualizing the microcirculation is attractive to apply in epidemiological studies and to monitor cardiovascular health from early age up to old age.
Systemic diseases that affect the circulation can result in progressive morphological changes in the retinal vasculature. For example, changes in the vessel calibers of retinal arteries and veins have been associated with hypertension, atherosclerosis, and increased risk of stroke and myocardial infarction. The vessel widths are derived using image analysis software and the width of the six largest arteries and veins are summarized in the Central Retinal Arteriolar Equivalent (CRAE) and the Central Retinal Venular Equivalent (CRVE). The latter features have been shown useful to study the impact of modifiable lifestyle and environmental cardiovascular disease risk factors.
The procedures to acquire fundus images and the analysis steps to obtain CRAE and CRVE are described. Coefficients of variation of repeated measures of CRAE and CRVE are less than 2% and within-rater reliability is very high. Using a panel study, the rapid response of the retinal vessel calibers to short-term changes in particulate air pollution, a known risk factor for cardiovascular mortality and morbidity, is reported. In conclusion, retinal imaging is proposed as a convenient and instrumental tool for epidemiological studies to study microvascular responses to cardiovascular disease risk factors.

Retinal image analysis is an unobtrusive procedure for visualizing the microcirculation. The impact of cardiovascular disease risk factors can result in changes of retinal vessel calibers. The procedures to acquire fundus images and the steps for calculating the vessel calibers are described.

眼底摄影作为一种方便的工具，研究微血管回应流行病学研究心血管疾病的风险因素

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an ...

Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation waiting list die without receiving an organ transplant or are delisted due to disease progression. Even after a successful transplantation, rejection and side effects of immunosuppression remain major concerns for graft survival and patient morbidity. 
Experimental animal research has been essential to the success of liver transplantation and still plays a pivotal role in the development of clinical transplantation practice. In particular, the porcine orthotopic liver transplantation model (OLTx) is optimal for clinically oriented research for its close resemblance to human size, anatomy, and physiology. 
Decompression of intestinal congestion during the anhepatic phase of porcine OLTx is important to guarantee reliable animal survival. The use of an active porto-caval-jugular shunt achieves excellent intestinal decompression. The system can be used for short-term as well as long-term survival experiments. The following protocol contains all technical information for a stable and reproducible liver transplantation model in pigs including post-operative animal care.

Experimental animal research plays a pivotal role in the development of clinical transplantation practice. The porcine orthotopic liver transplantation model (OLTx) closely resembles human conditions and is frequently used in clinically oriented research. The following protocol contains all information for a reliable porcine OLTx model using an active porto-caval-jugular shunt.

使用Active波尔图腔静脉分流术猪肝脏采购和原位移植技术

The success of liver transplantation has resulted in a dramatic organ shortage. Each year, a considerable number of patients on the liver transplantation ...

Automated Measurement of Pulmonary Emphysema and Small Airway Remodeling in Cigarette Smoke-exposed Mice

COPD is projected to be the third most common cause of mortality world-wide by 2020(1). Animal models of COPD are used to identify molecules that contribute to the disease process and to test the efficacy of novel therapies for COPD. Researchers use a number of models of COPD employing different species including rodents, guinea-pigs, rabbits, and dogs(2). However, the most widely-used model is that in which mice are exposed to cigarette smoke. Mice are an especially useful species in which to model COPD because their genome can readily be manipulated to generate animals that are either deficient in, or over-express individual proteins. Studies of gene-targeted mice that have been exposed to cigarette smoke have provided valuable information about the contributions of individual molecules to different lung pathologies in COPD(3-5). Most studies have focused on pathways involved in emphysema development which contributes to the airflow obstruction that is characteristic of COPD. However, small airway fibrosis also contributes significantly to airflow obstruction in human COPD patients(6), but much less is known about the pathogenesis of this lesion in smoke-exposed animals. To address this knowledge gap, this protocol quantifies both emphysema development and small airway fibrosis in smoke-exposed mice. This protocol exposes mice to CS using a whole-body exposure technique, then measures respiratory mechanics in the mice, inflates the lungs of mice to a standard pressure, and fixes the lungs in formalin. The researcher then stains the lung sections with either Gill&#8217;s stain to measure the mean alveolar chord length (as a readout of emphysema severity) or Masson&#8217;s trichrome stain to measure deposition of extracellular matrix (ECM) proteins around small airways (as a readout of small airway fibrosis). Studies of the effects of molecular pathways on both of these lung pathologies will lead to a better understanding of the pathogenesis of COPD.

The goal of this protocol is to provide automated methods to quantify chronic lung pathologies in a murine model of COPD. The protocol includes exposing mice to cigarette smoke (CS), measuring pulmonary function, inflating the lungs, and using morphometry methods to measure emphysema and small airway remodeling in mice.

肺气肿，小气道重塑的香烟烟雾暴露小鼠自动测量

COPD is projected to be the third most common cause of mortality world-wide by 2020(1).  Animal models of COPD are used to identify molecules that ...

Cardiac Catheterization in Mice to Measure the Pressure Volume Relationship: Investigating the Bowditch Effect

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these strategies is to examine their effects on heart function. There are several techniques to measure in vivo cardiac mechanics (e.g., echocardiography, pressure/volume relations, etc.). Compared to echocardiography, real-time left ventricular (LV) pressure/volume analysis via catheterization is more precise and insightful in assessing LV function. Additionally, LV pressure/volume analysis provides the ability to instantaneously record changes during manipulations of contractility (e.g., &#946;-adrenergic stimulation) and pathological insults (e.g., ischemia/reperfusion injury). In addition to the maximum (+dP/dt) and minimum (-dP/dt) rate of pressure change in the LV, an accurate assessment of LV function via several load-independent indexes (e.g., end systolic pressure volume relationship and preload recruitable stroke work) can be attained. Heart rate has a significant effect on LV contractility such that an increase in the heart rate is the primary mechanism to increase cardiac output (i.e., Bowditch effect). Thus, when comparing hemodynamics between experimental groups, it is necessary to have similar heart rates. Furthermore, a hallmark of many cardiomyopathy models is a decrease in contractile reserve (i.e., decreased Bowditch effect). Consequently, vital information can be obtained by determining the effects of increasing heart rate on contractility. Our and others data has demonstrated that the neuronal nitric oxide synthase (NOS1) knockout mouse has decreased contractility. Here we describe the procedure of measuring LV pressure/volume with increasing heart rates using the NOS1 knockout mouse model.

This article describes the measurement of murine left ventricular function via pressure/volume analysis at different heart rates.

心导​​管在小鼠测量压力体积的关系:调查的影响鲍迪奇

Animal models that mimic human cardiac disorders have been created to test potential therapeutic strategies. A key component to evaluating these ...

Murine Prostate Micro-dissection and Surgical Castration

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has been used as a surrogate for discoveries in human prostate development and disease. Prostate micro-dissection allows consistent study of lobe-specific prostate anatomy, histology, and cellular characteristics in the absence of contamination of other tissues. Testosterone affects prostate development and disease. Androgen deprivation therapy is a common treatment for prostate cancer patients, but many prostate tumors become castration-resistant. Surgical castration of mouse models allows for the study of castration resistance and other facets of hormonal biology on the prostate. This procedure can be coupled with testosterone reintroduction, or hormonal regeneration of the prostate, a powerful method to study stem cell lineages in the prostate. Together, prostate micro-dissection and surgical castration opens up a multitude of opportunities for robust and consistent research of prostate development and disease. This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse.

This manuscript describes the protocols for prostate micro-dissection and surgical castration in the laboratory mouse. We also depict representative results produced by these protocols. Finally, we discuss the advantages and utilization of these protocols.

鼠前列腺微解剖和手术去势

Mouse models are used extensively to study prostate cancer and other diseases. The mouse is an excellent model with which to study the prostate and has ...

Generation of Organ-conditioned Media and Applications for Studying Organ-specific Influences on Breast Cancer Metastatic Behavior

Breast cancer preferentially metastasizes to the lymph node, bone, lung, brain and liver in breast cancer patients. Previous research efforts have focused on identifying factors inherent to breast cancer cells that are responsible for this observed metastatic pattern (termed organ tropism), however much less is known about factors present within specific organs that contribute to this process. This is in part because of a lack of in vitro model systems that accurately recapitulate the organ microenvironment. To address this, an ex vivo model system has been established that allows for the study of soluble factors present within different organ microenvironments. This model consists of generating conditioned media from organs (lymph node, bone, lung, and brain) isolated from normal athymic nude mice. The model system has been validated by demonstrating that different breast cancer cell lines display cell-line specific and organ-specific malignant behavior in response to organ-conditioned media that corresponds to their in vivo metastatic potential. This model system can be used to identify and evaluate specific organ-derived soluble factors that may play a role in the metastatic behavior of breast and other types of cancer cells, including influences on growth, migration, stem-like behavior, and gene expression, as well as the identification of potential new therapeutic targets for cancer. This is the first ex vivo model system that can be used to study organ-specific metastatic behavior in detail and evaluate the role of specific organ-derived soluble factors in driving the process of cancer metastasis.

This manuscript describes an ex vivo model system comprised of organ-conditioned media derived from the lymph node, bone, lung, and brain of mice. This model system can be used to identify and study organ-derived soluble factors and their effects on the organ tropism and metastatic behavior of cancer cells.

器官空调，媒体和应用的产生对乳腺癌转移行为研究器官特异性的影响

Breast cancer preferentially metastasizes to the lymph node, bone, lung, brain and liver in breast cancer patients. Previous research efforts have focused ...

Isolation and Cultivation of Mandibular Bone Marrow Mesenchymal Stem Cells in Rats

Here we present an efficient method for isolating and culturing mandibular bone marrow mesenchymal stem cells (mBMSCs) in vitro to rapidly obtain numerous high-quality cells for experimental requirements. mBMSCs could be widely used in therapeutic applications as tissue engineering cells in case of craniofacial diseases and cranio-maxillofacial regeneration in the future due to the excellent self-renewal ability and multi-lineage differentiation potential. Therefore, it is important to obtain mBMSCs in large numbers.
In this study, bone marrow was flushed from the mandible and primary mBMSCs were isolated through whole bone marrow adherent cultivation. Furthermore, CD29+CD90+CD45&#8722; mBMSCs were purified through fluorescent cell sorting. The second generation of purified mBMSCs were used for further study and displayed potential in differentiating into osteoblasts, adipocytes, and chondrocytes. Utilizing this in vitro model, one can obtain a high number of proliferative mBMSCs, which may facilitate the study of the biological characteristics, the subsequent reaction to the microenvironment, and other applications of mBMSCs.

This article presents a method that combines whole bone marrow adherence and flow cytometry sorting for isolating, cultivating, sorting, and identifying bone marrow mesenchymal stem cells from rat mandibles.

大鼠骨髓骨髓的隔离和培养

Here we present an efficient method for isolating and culturing mandibular bone marrow mesenchymal stem cells (mBMSCs) in vitro to rapidly obtain numerous ...

Operating Transverse Aortic Constriction with Absorbable Suture to Obtain Transient Myocardial Hypertrophy

Based on twice transverse aortic constrictions (TACs) in mice, it is proved that myocardial hypertrophic preconditioning (MHP) could attenuate cardiomyocyte hypertrophy and slow down progression to heart failure. For novices, however, the MHP model is usually quite difficult to establish because of the technical obstacles in ventilator operation, opening the chest repeatedly, and bleeding caused by debanding. To facilitate this model, to increase the surgical success rate and to reduce the incidence of bleeding, we switched to absorbable sutures for the first TAC combing with a ventilator-free technique. Using a 2-week absorbable suture, we demonstrated that this procedure could cause significant myocardial hypertrophy in 2 weeks; and 4 weeks after surgery, myocardial hypertrophy was almost completely regressed to the baseline. Using this protocol, the operators could master the MHP model easily with a lower operation mortality.

This protocol presents an improved method to obtain transient myocardial hypertrophy with absorbable suture, simulating left ventricular hypertrophy decrease after removing pressure overload. It could be valuable for the studies on myocardial hypertrophic preconditioning.

操作横向主动脉收缩与可吸收的缝合获得瞬时心肌萎缩

Based on twice transverse aortic constrictions (TACs) in mice, it is proved that myocardial hypertrophic preconditioning (MHP) could attenuate ...

3D Imaging of PDL Collagen Fibers during Orthodontic Tooth Movement in Mandibular Murine Model

Orthodontic tooth movement is a complex biological process of altered soft and hard tissue remodeling as a result of external forces. In order to understand these complex remodeling processes, it is critical to study the tooth and periodontal tissues within their 3D context and therefore minimize any sectioning and tissue artefacts. Mouse models are often utilized in developmental and structural biology, as well as in biomechanics due to their small size, high metabolic rate, genetics and ease of handling. In principle this also makes them excellent models for dental related studies. However, a major impediment is their small tooth size, the molars in particular. This paper is aimed at providing a step by step protocol for generating orthodontic tooth movement and two methods for 3D imaging of the periodontal ligament fibrous component of a mouse mandibular molar. The first method presented is based on a micro-CT setup enabling phase enhancement&#160;imaging of fresh collagen tissues. The second method is a bone clearing method using ethyl cinnamate that enables imaging through the bone without sectioning and preserves endogenous fluorescence. Combining this clearing method with reporter mice like Flk1-Cre;TdTomato provided a first of its kind opportunity to image the 3D vasculature in the PDL and alveolar bone.

We present a protocol for generating orthodontic tooth movement in mice and methods for 3D visualization of the collagen fibers and blood vessels of periodontal ligament without sectioning.

环境毒物暴露大鼠下颌切牙釉质器官的显微解剖

摘要

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此视频中的章节

测量和制止在弱视治疗

眼底摄影作为一种方便的工具，研究微血管回应流行病学研究心血管疾病的风险因素

使用Active波尔图腔静脉分流术猪肝脏采购和原位移植技术

肺气肿，小气道重塑的香烟烟雾暴露小鼠自动测量

心导管在小鼠测量压力体积的关系:调查的影响鲍迪奇

鼠前列腺微解剖和手术去势

器官空调，媒体和应用的产生对乳腺癌转移行为研究器官特异性的影响

大鼠骨髓骨髓的隔离和培养

操作横向主动脉收缩与可吸收的缝合获得瞬时心肌萎缩

曼迪布勒穆林模型中矫形牙齿运动期间 PDL 胶原蛋白纤维的 3D 成像

环境毒物暴露大鼠下颌切牙釉质器官的显微解剖

摘要

探索更多视频

此视频中的章节

测量和制止在弱视治疗

眼底摄影作为一种方便的工具，研究微血管回应流行病学研究心血管疾病的风险因素

使用Active波尔图腔静脉分流术猪肝脏采购和原位移植技术

肺气肿，小气道重塑的香烟烟雾暴露小鼠自动测量

心导​​管在小鼠测量压力体积的关系:调查的影响鲍迪奇

鼠前列腺微解剖和手术去势

器官空调，媒体和应用的产生对乳腺癌转移行为研究器官特异性的影响

大鼠骨髓骨髓的隔离和培养

操作横向主动脉收缩与可吸收的缝合获得瞬时心肌萎缩

曼迪布勒穆林模型中矫形牙齿运动期间 PDL 胶原蛋白纤维的 3D 成像

心导管在小鼠测量压力体积的关系:调查的影响鲍迪奇