作者感谢丽莎 Pichl 出色的技术援助。

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作者没有什么可透露的。

从猪动物模型中产生的生物库样本收集需要强健的技术和协议, 以确定器官/组织体积, 可再现的代表性, 多余的组织样本, 适合广泛的不同的分析方法, 并对抽样剖面的随机化进行定量视学分析。本条所述方法适用于猪器官和组织的大小, 并已开发出有效满足这些要求的2,7。它们基于公认的方法论原则, 并在不同的发布研究中证明了它们的实用性<sup class="xr...

在翻译医学中, 猪越来越普遍地使用作为大动物模型1,2,3,4,5, 由于猪的几个有利相似性和人体解剖和生理学, 以及现有的分子生物学方法, 允许为多种疾病条件生成量身定做的基因改良猪模型1,4。
然而, 与啮齿动物模型相比, 可以随时为实验提供的不同猪模型的动物数量是有限的。这是由于猪的世代间隔大约一年, 并且为猪模型和畜牧业的世代需要的财政和时间密集的努力。因此, 猪模型的个体动物, 以及可以从这些猪身上生成的样本, 都非常有价值, 特别是如果转基因猪模型和/或长期实验性问题 (例如, 晚期并发症的慢性病) 在老年个体检查2,6,7。
在任何研究过程中, 在研究的初步实验设计中尚未排定的额外分析的表现可能后来变成相关的,例如, 以解决由于先前发现的不同问题意想不到的发现。如果没有合适的样品进行这种额外的试验, 不成比例可能需要花费高昂的成本和耗费大量时间的开支来产生更多的猪和组织样本。为应对这种可能性, 生物库收集不同器官、组织或生物液体的保守备份样本, 在数量上和质量上都适合广泛的后续分析, 被认为是重要的方法2,6,7。从猪的动物模型中获得最佳的好处, 充分的生物库样本的可用性也提供了独特的可能性, 在同一样本材料上执行广泛的不同分析方法在一个多器官水平上, 在同一单个动物,例如, 通过在研究网络2、6、7中组织的不同工作组的科学家分发样本。此外, biobanking 的 "前瞻性" 抽样策略也有助于减少研究中需要的动物数量。猪模型 biobanking 的优势最近已在多器官, multiomics 研究, 检查器官交叉谈话的基因改良猪模型长期糖尿病, 使用标本从慕尼黑米迪猪生物库2。
有一些强制性要求生物库样品必须普遍遵守, 以建立可靠性和解读的结果, 随后进行分析。必须重现性生成样本, 并且它们必须具有代表性, 即, 充分反映了从7中抽取样本的组织/器官感兴趣的形态学和分子特征. 为了适合广泛的下游分析类型, 样品必须采取足够的数量和处理, 根据要求 (包括时间和温度条件) 的不同分析方法, 包括描述性病理学分析, 如 cryohistology, 石蜡和整形组织学, 免疫组化,原位杂交, 超微结构的电子显微分析, 临床实验室诊断分析, 以及分子分析 DNA, RNA, 蛋白质和代谢物。
通过定量视学分析, 允许对多种不同的定量形态学参数 (如数字、体积、长度或不同组织结构的表面积) 进行评估, 随机断面平面各器官/组织的组织学样本需要准备7,8,9,10,11。在定量形态学研究中, 对组织、器官或器官室总容积的精确测定, 样本取自 (即, 参考空间) 至关重要7,9,12计算相应器官、组织或有机体中相关参数的绝对数量。最终, 在组织学切片的制备过程中, 嵌入相关组织收缩的影响必须确定并考虑到13。因此, 以往研究中的定量视学分析, 特别是存档样本 (固定组织样本、嵌入组织块、组织学切片、等等) 有时会受到严重限制, 甚至是不可能的12,特别是如果没有执行有关的器官/组织的容量, 如果没有充分的抽样设计适用于保证代表性样品, 如果数量和数量的可利用的单独样品是不足的, 或者, 如果处理样本与感兴趣的定量形态学参数的估计不相容。由于各种可能的影响因素, 档案样本材料对不同数量形态参数分析的适用性不能明确地得到回答, 而是取决于对个别案件的仔细评估。
因此, 由于样品的位置、大小、数量、加工、修剪方向和方向等因素对后续分析的结果有潜在影响, 因此这些因子具有重要意义, 必须在任何研究的实验设计中加以考虑。关于这些方面和猪解剖学的特点, 最近建立了适合猪动物模型的全面、详细、大规模的抽样准则, 为标准化、可重现性提供了有力的参考。, 并高效生成冗余、经过充分处理的高质量样品, 来自50多个不同的猪器官和组织6,7。
本文所示的方法学描述和视频教程提供了详细的、说明性的、可理解的、循序渐进的指导, 以实现各种容量技术的实际性能、猪组织取样和器官, 并处理组织样品的不同下游分析方法。精选的技术包括根据阿基米德和卡瓦列里的原理确定器官/组织体积和密度的方法9, 包括确定与所涉组织的三维收缩的尺寸嵌入到不同的嵌入媒体14在组织学检查过程中, 应用实际体积加权系统随机抽样方法, 对不同后续的取样组织标本进行处理。分析7,8,9,15, 并为潜在的定量视学分析生成适当的面向和处理的样本7,8, 9,10,11。除了在猪生物库项目中的应用, 所展示的方法一般适用于检查器官/组织的定量历史形态特性的所有研究。此外, 系统随机抽样设计特别有利于在实验中产生代表性样品, 方法是利用分子分析法检测在各种器官和组织。
下段简要介绍了这些方法, 而它们的实际性能则在 "协议" 部分中进行了说明。
器官/组织体积的测定 在几个实验环境中, 器官重量和体积的测定是很重要的, 因为这些因素可能表明变化, 可能与实验性研究的兴趣因素有关。一个器官/组织的总体积也通常需要计算绝对数量参数, (例如, 总细胞数), 从 stereologically 估计的数字体积密度 (即, 每个体积单位的细胞数组织)7,12。除了使用复杂的技术设备 (如计算机断层扫描) 技术外, 基本上还有三种常用的方法来确定器官或组织的绝对体积。一个器官的容量可以由 "直接容积测量" 决定根据阿基米德的原则,即, 测量水的容量或盐水由结构完全地淹没时偏移。然而, 对于比较大的猪器官, 这些方法是不切实际的, 容易不精确, 因为它们需要非常大的容积/测量烧瓶。更方便的是, 一个器官/组织的体积可以计算从它的重量和密度7,12,16, 可以有效地确定使用 "淹没方法"7,12 ,16 (协议步骤 1.1)。器官/组织体积也可以根据 "卡瓦列里原则" (1598–1647) 使用容量方法估计。简单地说, 卡瓦列里原则规定, 如果两个物体在平行于地面平面的平面上切片, 而在与地面平面相对应的距离上切割两个物体的剖面轮廓有相同的区域, 则两个物体有相同的音量。因此, 任意形状物体的体积可以被估计为其截面剖面区域的乘积, 平行, 同样遥远的剖面平面和截面平面之间的距离。这是可理解的与以下类比: 考虑两个栈组成相同数量的相同的硬币是并排放置, 一个栈与硬币有序堆放在一起产生圆柱形的硬币栈, 和其他用离中心位置的硬币堆叠的硬币 (图 3A)。虽然两个硬币栈的形状是不同的, 它们的体积是相同的, 因为硬币的面积在两个栈的相应级别 (即, 平行截面轮廓的区域通过两个硬币栈在相同的距离切割地面) 是相同的。使用卡瓦列里原理估计猪器官和组织的体积7,12,15在步骤1.2 中描述。
组织学埋植相关组织收缩程度的测定 在对组织学组织切片测量的几种定量形态学参数进行分析时, 必须确定并考虑组织处理过程中嵌入相关组织收缩对组织学的影响。嵌入相关组织收缩的程度可能是可变的, 并且取决于组织、处理和嵌入介质8,13,17,18,19。通常, 组织样本体积的嵌入相关变化 (即, 主要是收缩) 发生在所有三维空间中, 因此影响由定量视学分析估计的所有维度参数8.基本上, 嵌入相关组织收缩的程度, 表示为线性组织收缩因子 (fS), 可以估计如步骤1.3 所示。用于校正 (收缩敏感) 定量形态学参数14。
器官/组织体积加权系统随机抽样 为建立生物库收集猪器官/组织样本, 体积加权系统随机抽样方法, 如步骤2所述, 证明是切实可行的, 节省时间, 和有效的技术, 以产生代表,多用途组织样本7,8,9,15。
定量视学分析的各向同性均匀随机剖面和垂直均匀随机剖面的生成 生物库组织样本需要适用于多种不同的定量视学分析方法, 用于估计在没有充分制备标本的情况下无法确定的最大参数。几乎所有定量视学参数都可以用 "各向同性 (独立) 均匀随机 (产学研)" 部分 "8,9" 来确定。在产学研切片中, 组织样品断面平面的三维方向是随机的。这可以通过随机化组织样本的位置相对于截面平面的位置, 如应用于 "Isector" 方法11 (协议步骤 3.1), 或通过随机化截面平面相对于组织样本, 如 "定位装置" 方法 10 (协议步骤 3.2).在组织样本中, 如皮肤或黏膜标本显示自然存在, 或定义和正确地可识别的垂直轴, 准备 "垂直均匀随机 (VUR) 部分" (协议步骤 3.3.) 严格地切片在他们的飞机垂直轴有利8,20。为了全面论述产学研/VUR 抽样的理论基础, 并对潜在的下游定量视学分析进行了综合讨论, 读者可以参考生活中定量视学教科书。科学8,9。

<table><tbody><tr><td>Agar</td><td>Carl Roth GmbH, Germany</td><td>Agar (powder), Cat.: 5210.3</td><td>Dissolve approximately 1 g of agar in 10 ml cold water in a glass or plastic beaker, heat in microwave-oven at 700 W, boil the solution twice with rigorous stirring. Cast into mold while still warm and let solidify. Caution: While handling with hot liquid agar, wear protective goggles and gloves.</td></tr><tr><td>Caliper</td><td>Hornbach Baumarkt GmbH, Bornheim, Germany</td><td>Schieblehre Chrom/Vernickelt 120 mm Cat.: 3664902</td><td>Any kind of caliper (mechanical or electronic) will do as well.</td></tr><tr><td>Casting molds (metal)</td><td>Engelbrecht Medizin &amp;amp; Labortechnik, Ederm&amp;uuml;nde, Germany</td><td>Einbettsch&amp;auml;lchen aus Edelstahl, 14 x 24 x 5 mm, Cat.: 14302b</td><td>Any other kind of metal casting mold used for paraffin-embedding will do as well.</td></tr><tr><td>Copy templates of cross grids (5mm - 6 cm)</td><td>n.a.</td><td>n.a.</td><td>Copy templates of cross grids (5mm - 6 cm) are provided in the supplemental data file of &lt;strong&gt;Albl et al.&lt;/strong&gt;&amp;nbsp; Toxicol Pathol. 44, 414-420, doi: 10.1177/0192623316631023 (2016)</td></tr><tr><td>Copy templates of equiangular and cosine-weighted circles</td><td>n.a.</td><td>n.a.</td><td>Copy templates of equiangular and cosine-weighted circles are provided in &lt;strong&gt;Nyengaard &amp;amp; Gundersen&lt;/strong&gt;. Eur Respir Rev. 15, 107-114, doi: 10.1183/09059180.00010101 (2006) and in &lt;strong&gt;Gundersen et al.&lt;/strong&gt; Stereological Principles and Sampling Procedures for Toxicologic Pathologists. In: Haschek and Rousseaux&amp;acute;s Handbook of Toxicologic Pathology. 3rd ed, 215-286, ISBN: 9780124157590 (2013).</td></tr><tr><td>Foldback clamps (YIHAI binder clips, 15 mm and 19 mm)</td><td>Ningbo Tianhong Stationery Co ltd., China</td><td>Y10006 and Y10005</td><td>Any other type of standard office foldback clamps will do as well.</td></tr><tr><td>Forceps (anatomical)</td><td>NeoLab Migge GmbH, Heidelberg, Germany</td><td>neoLab Standard -Pinzette 130 mm, anatomisch, rund, Cat.: 1-1811</td><td>Any type of anatomical forceps will do.</td></tr><tr><td>Formaldehyde-solution 4%</td><td>SAV-Liquid Produktion GmbH, Flintsbach, Germany</td><td>Formaldehyd 37/40 %, Cat.: 1000411525005</td><td>Dilute to 4% from concentrated solution. Buffer to neutral pH. Wear appropriate eye-, hand- and respiratory protection. Process tissue samples fixed in formaldehyde solution under an exhaust hood and wear protective goggles and laboratory gloves.</td></tr><tr><td>Graph paper (for calibration)</td><td>B&amp;uuml;romarkt B&amp;ouml;ttcher AG, Jena, Germany. www.bueromarkt-ag.de</td><td>Penig Millimeterpapier A4, Cat.: 2514</td><td>Any type of graph paper (scaled in millimeter) will do.</td></tr><tr><td>Laboratory beakers (5ml, 10 ml, 50 ml, 100 ml)</td><td>NeoLab Migge GmbH, Heidelberg, Germany</td><td>Becherglas SIMAX&amp;reg; , niedrige Form, Borosilikatglas 3.3 Cat.: E-1031, E-1032, E-1035, E-1036</td><td>Any kind of glass- or plastic beakers of 5 &amp;ndash; 100 ml volume will do.</td></tr><tr><td>Laboratory scale(s)</td><td>Mettler Toledo GmbH, Gie&amp;szlig;en, Germany</td><td>PM6000</td><td>Any standard laboratory scales with measuring ranges between 0.1 mg to approximately 20 g, respectively between 100 mg to approximately 500 g will do</td></tr><tr><td></td><td>Sartorius AG, G&amp;ouml;ttingen, Germany</td><td>BP61S</td><td></td></tr><tr><td>Microtome blades</td><td>Engelbrecht Medizin &amp;amp; Labortechnik, Ederm&amp;uuml;nde, Germany</td><td>FEATHER Microtome blasdes S35, Cat.:14700</td><td>Any kind of single-use microtome blades will do.</td></tr><tr><td>Morphometry/planimetry software/system</td><td>National Institute of Health (NIH)</td><td>ImageJ</td><td>Download from https://www. imagej.nih.gov/ij/ (1997).</td></tr><tr><td>Zeiss-Kontron, Eching, Germany</td><td>VideoplanTM image analysis system</td><td>Out of stock</td><td></td></tr><tr><td>Photo camera</td><td>Nikon</td><td>D40</td><td>Any kind of digital photocamera that can be mounted to a tripod&amp;nbsp; will do.</td></tr><tr><td>Plastic transparencies</td><td>Avery Zweckform GmbH, Oberlaindern, Germany</td><td>Laser Overhead-Folie DINA4 Cat.:&amp;nbsp; 3562</td><td>Any (laser)-printable plastic transparency will do.</td></tr><tr><td>Random number tables</td><td>n.a.</td><td>n.a.</td><td>Random number tables can conveniently be generated (with defined numbers of random numbers and within defined intervals), using random number generators, such as: https://www.random.org/</td></tr><tr><td>Razor blades</td><td>Plano GmbH, Wetzlar, Germany</td><td>T5016</td><td>Any kind of razor blades will do.</td></tr><tr><td>Ruler</td><td>B&amp;uuml;romarkt B&amp;ouml;ttcher AG, Jena, Germany. www.bueromarkt-ag.de</td><td>Office-Point Lineal 30 cm, Kunststoff, transparent, Cat.: ln30</td><td>Any kind of cm-mm-scaled ruler will do as well.</td></tr><tr><td>Saline (0.9%)</td><td>Carl Roth GmbH, Germany</td><td>Natriumchlorid, &gt;99% Cat.: 0601.1</td><td>To prepare 0.9% saline, dissolve 9 g NaCl in 1000 ml of distilled water at 20&amp;deg;C.</td></tr><tr><td>Scalpel blades</td><td>Aesculap AG &amp;amp; Co KG, Tuttlingen, Germany</td><td>BRAUN Surgical blades N&amp;deg;22</td><td>Any kind of scalpel blades will do.</td></tr><tr><td>Scanner</td><td>Hewlett-Packard</td><td>hp scanjet 7400c</td><td>Any type of standard office scanner capable of scanning with resolutions from 150-600 dpi will do.</td></tr><tr><td>Slicing devices</td><td>n.a.</td><td>n.a.</td><td>Examples forself constructed slicing devices can be found in &lt;strong&gt;Knust, et al.&lt;/strong&gt; Anatomical record. 292, 113-122, doi: 10.1002/ar.20747 (2009) and in the supplemental data file of&lt;strong&gt; Albl et al.&amp;nbsp; &lt;/strong&gt;Toxicol Pathol. 44, 414-420, doi: 10.1177/0192623316631023 (2016).</td></tr><tr><td>Spherical casting molds (e.g., in 25.5 mm diameter)</td><td>Pralinen-Zutaten.de, Windach, Germany</td><td>Pralinen-Hohlkugeln Vollmilch, 25.5 mm</td><td>Spherical casting molds can as well be be self-constructed, or obtained from other confectioner suppliers (for for pralines). The casting molds indicated here are actually the package/wrapping of hollow pralines bodies (first eat the pralines and then use the package for generation of i-sector sections)</td></tr><tr><td>Thin wire</td><td>Basteln &amp;amp; Hobby Schobes, Stra&amp;szlig;furth, Germany. www,bastel-welt.de</td><td>Messingdraht (0.3 mm) Cat.: 216464742</td><td>Any other kind of thin wire will also do.</td></tr><tr><td>Tissue paper</td><td>NeoLab Migge GmbH, Heidelberg, Germany</td><td>Declcate Task Wipes-White, Cat.: 1-5305</td><td>Any other kind of laboratory tissue paper will do as well.</td></tr><tr><td>Waterproof pen</td><td>Staedler Mars GmbH &amp;amp; Co KG, N&amp;uuml;rnberg, Grmany</td><td>Lumocolor permanent 313, 0.4 mm, S, black, Cat.: 313-2</td><td>Any other kind of waterproof pen will do as well.</td></tr></tbody></table>

sampling strategies and processing of biobank tissue samples from porcine biomedical models

在翻译医学研究中, 猪模型逐渐变得越来越受欢迎。考虑到个别动物的高价值, 特别是基因改良猪模型, 以及这些模型的可用动物数量有限, 建立了适当加工的组织样本 (生物库), 适合于广泛的后续分析方法, 包括没有在取样时间点指定的分析, 代表了充分利用模型的平移值的有意义的方法。关于猪解剖学的特殊性, 最近建立了综合指导方针, 以标准化生成代表性的, 高质量的样品从不同的猪器官和组织。这些指导原则是结果重现性的必要条件, 也是不同研究和调查人员之间的可比。基本数据的记录, 例如器官重量和体积、取样位置的确定和要生成的组织样本数量, 以及它们的方向、大小、加工和修剪方向, 都是相关因素。确定标本的概化和可用性的分子, 定性, 定量形态学分析。本文介绍了猪组织中具有代表性的、多用途的生物库标本最重要的生产技术。这里描述的方法包括测定器官/组织体积和密度, 用点计数法对实质器官进行容积加权系统随机抽样程序的应用, 确定组织收缩程度与样本的组织学嵌入有关, 并生成随机定向样本进行定量视学分析, 如由 "定位装置" 和 "Isector" 方法产生的各向同性均匀随机 (产学研) 剖面, 以及垂直均匀随机 (VUR) 部分。

浸没法测定组织/器官密度
图 12A-b显示使用步骤 1.1 (图 1,图 2) 中描述的淹没技术来确定猪肾脏密度和体积的代表性。其他猪器官和组织的密度测量的更有代表性的结果载于表 2。"猪生物医学模型的组织取样指南"7显示了...

Watch this Scientific Journal Video about Sampling Strategies and Processing of Biobank Tissue Samples from Porcine Biomedical Models at JoVE.com

Sampling Strategies and Processing of Biobank Tissue Samples from Porcine Biomedical Models

猪生物医学模型生物库组织样品的取样策略与处理

在生物库项目的下游分析中, 展示了猪动物模型代表性组织样本生成方法的实际应用和性能, 包括容量、系统随机抽样、组织样品的鉴别处理, 用于定性和定量的形态学和分子分析类型。

In translational medical research, porcine models have steadily become more popular. Considering the high value of individual animals, particularly of ...

sampling-strategies-processing-biobank-tissue-samples-from-porcine

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

15.6K 次观看.  LMU Munich. 在生物库项目的下游分析中, 展示了猪动物模型代表性组织样本生成方法的实际应用和性能, 包括容量、系统随机抽样、组织样品的鉴别处理, 用于定性和定量的形态学和分子分析类型。

视频: Sampling Strategies and Processing of Biobank Tissue Samples from Porcine Biomedical Models

A Coupled Experiment-finite Element Modeling Methodology for Assessing High Strain Rate Mechanical Response of Soft Biomaterials

This study offers a combined experimental and finite element (FE) simulation approach for examining the mechanical behavior of soft biomaterials (e.g. brain, liver, tendon, fat, etc.) when exposed to high strain rates. This study utilized a Split-Hopkinson Pressure Bar (SHPB) to generate strain rates of 100-1,500 sec-1. The SHPB employed a striker bar consisting of a viscoelastic material (polycarbonate). A sample of the biomaterial was obtained shortly postmortem and prepared for SHPB testing. The specimen was interposed between the incident and transmitted bars, and the pneumatic components of the SHPB were activated to drive the striker bar toward the incident bar. The resulting impact generated a compressive stress wave (i.e. incident wave) that traveled through the incident bar. When the compressive stress wave reached the end of the incident bar, a portion continued forward through the sample and transmitted bar (i.e. transmitted wave) while another portion reversed through the incident bar as a tensile wave (i.e. reflected wave). These waves were measured using strain gages mounted on the incident and transmitted bars. The true stress-strain behavior of the sample was determined from equations based on wave propagation and dynamic force equilibrium. The experimental stress-strain response was three dimensional in nature because the specimen bulged. As such, the hydrostatic stress (first invariant) was used to generate the stress-strain response. In order to extract the uniaxial (one-dimensional) mechanical response of the tissue, an iterative coupled optimization was performed using experimental results and Finite Element Analysis (FEA), which contained an Internal State Variable (ISV) material model used for the tissue. The ISV material model used in the FE simulations of the experimental setup was iteratively calibrated (i.e. optimized) to the experimental data such that the experiment and FEA strain gage values and first invariant of stresses were in good agreement.

The current study prescribes a coupled experiment-finite element simulation methodology to obtain the uniaxial dynamic mechanical response of soft biomaterials (brain, liver, tendon, fat, etc.). The multiaxial experimental results that arose because of specimen bulging obtained from Split-Hopkinson Pressure Bar testing were rendered to a uniaxial true stress-strain behavior when simulated through iterative optimization of the finite element analysis of the biomaterial.

一个耦合试验，有限元建模方法以评估生物材料软的高应变率力学响应

This study offers a combined experimental and finite element (FE) simulation approach for examining the mechanical behavior of soft biomaterials (e.g. ...

科研

生物工程

Creation and Maintenance of a Living Biobank - How We Do It

In light of the growing knowledge about the inter-individual properties and heterogeneity of cancers, the emerging field of personalized medicine requires a platform for preclinical research. Over recent years, we have established a biobank of colorectal and pancreatic cancers comprising of primary tumor tissue, normal tissue, sera, isolated peripheral blood lymphocytes (PBL), patient-derived xenografts (PDX), as well as primary and secondary cancer cell lines. Since original tumor tissue is limited and the establishment rate of primary cancer cell lines is still relatively low, PDX allow not only the preservation and extension of the biobank but also the generation of secondary cancer cell lines. Moreover, PDX-models have been proven to be the ideal in vivo model for preclinical drug testing. However, biobanking requires careful preparation, strict guidelines and a well attuned infrastructure. Colectomy, duodenopancreatectomy or resected metastases specimens are collected immediately after resection and transferred to the pathology department. Respecting priority of an unbiased histopathological report, at the discretion of the attending pathologist who carries out the dissections, small tumor pieces and non-tumor tissue are harvested. 
Necrotic parts are discarded and the remaining tumor tissue is cut into small, identical cubes and cryopreserved for later use. Additionally, a small portion of the tumor is minced and strained for primary cancer cell culture. Additionally, blood samples drawn from the patient pre- and postoperatively, are processed to obtain serum and PBLs. For PDX engraftment, the cryopreserved specimens are defrosted and implanted subcutaneously into the flanks of immunodeficient mice. The resulting PDX closely recapitulate the histology of the "donor" tumors and can be either used for subsequent xenografting or cryopreserved for later use. In the following work, we describe the individual steps of creation, maintenance and administration of a large biobank of colorectal and pancreatic cancer. Moreover, we highlight the crucial details and caveats associated with biobanking.

In the following work, we describe the consecutive steps necessary for the establishment of a large biobank of colorectal and pancreatic cancer.

活生物库的创建和维护 - 我们如何做到这一点

In light of the growing knowledge about the inter-individual properties and heterogeneity of cancers, the emerging field of personalized medicine requires ...

癌症研究

Biobank for Translational Medicine: Standard Operating Procedures for Optimal Sample Management

Biobanks are key research infrastructures aimed at the collection, storage, processing, and sharing of high-quality human biological samples and associated data for research, diagnosis, and personalized medicine. The Biobank for Translational and Digital Medicine Unit at the European Institute of Oncology (IEO) is a landmark in this field. Biobanks collaborate with clinical divisions, internal and external research groups, and industry, supporting patients' treatment and scientific progress, including innovative diagnostics, biomarker discovery, and clinical trial design. Given the central role of biobanks in modern research, biobanking standard operating procedures (SOPs) should be extremely precise. SOPs and controls by certified specialists ensure the highest quality of samples for the implementation of science-based, diagnostic, prognostic, and therapeutic personalized strategies. However, despite numerous efforts to standardize and harmonize biobanks, these protocols, which follow a strict set of rules, quality controls, and guidelines based on ethical and legal principles, are not easily accessible. This paper presents the biobank standard operating procedures of a large cancer center.

Biobanks are crucial resources for biomedical research and the Biobank for Translational and Digital Medicine Unit at the European Institute of Oncology is a model in this field. Here, we provide a detailed description of biobanks' standard operating procedures for the management of different types of human biological samples.

转化医学生物样本库:优化样品管理的标准操作程序

Biobanks are key research infrastructures aimed at the collection, storage, processing, and sharing of high-quality human biological samples and ...

前列腺切除术标本标测和解剖方案

Enhancing Prostate Tumor Biobanking Reliability with Improved Sampling Technique and Histological Characterization

Acquiring fresh and well-characterized tumor tissue samples is critical for conducting high-quality &#34;omics&#34; studies. However, it can be particularly challenging in the context of prostate cancer (PC) due to the unique nature of this organ and the high heterogeneity associated with this tumor. On the other hand, histopathologically characterizing samples before their storage without causing significant tissue alterations is also an intriguing challenge. In this context, we present a new method for acquiring, mapping, characterizing, and micro-dissecting resected prostate tissue based on anatomopathological criteria.
Unlike previously published protocols, this method reduces the time required for histopathological analysis of the prostate specimen without compromising its structure, which is crucial for assessing surgical margins. Furthermore, it enables the delineation and micro-macro dissection of fresh prostate tissue samples, with a focus on histological tumor areas defined by pathological criteria such as Gleason score, precursor lesions (high-grade prostatic intraepithelial neoplasia - PIN), and inflammatory lesions (prostatitis). These samples are then stored in a Biobank for subsequent research analyses.

作者聚焦:通过改进组织取样和生物样本库推进前列腺癌研究

This protocol describes a method for facilitating the collection of samples from radical prostatectomy specimens. The goal is to map, characterize, and micro-macro dissect tissue samples from the specimens based on anatomopathological criteria before storing them in a Biobank.

通过改进采样技术和组织学表征提高前列腺肿瘤生物样本库的可靠性

Acquiring fresh and well-characterized tumor tissue samples is critical for conducting high-quality &#34;omics&#34; studies. However, it can be ...

Characterization of Blood Outgrowth Endothelial Cells (BOEC) from Porcine Peripheral Blood

The endothelium is a dynamic integrated structure that plays an important role in many physiological functions such as angiogenesis, hemostasis, inflammation, and homeostasis. The endothelium also plays an important role in pathophysiologies such as atherosclerosis, hypertension, and diabetes. Endothelial cells form the inner lining of blood and lymphatic vessels and display heterogeneity in structure and function. Various groups have evaluated the functionality of endothelial cells derived from human peripheral blood with a focus on endothelial progenitor cells derived from hematopoietic stem cells or mature blood outgrowth endothelial cells (or endothelial colony-forming cells). These cells provide an autologous resource for therapeutics and disease modeling. Xenogeneic cells may provide an alternative source of therapeutics due to their availability and homogeneity achieved by using genetically similar animals raised in similar conditions. Hence, a robust protocol for the isolation and expansion of highly proliferative blood outgrowth endothelial cells from porcine peripheral blood has been presented. These cells can be used for numerous applications such as cardiovascular tissue engineering, cell therapy, disease modeling, drug screening, studying endothelial cell biology, and in vitro co-cultures to investigate inflammatory and coagulation responses in xenotransplantation.

Characterization of Blood Outgrowth Endothelial Cells BOEC from Porcine Peripheral Blood

猪外周血血液生长内皮细胞（BOEC）的表征

The endothelium is a dynamic integrated structure that plays an important role in many physiological functions such as angiogenesis, hemostasis, ...

Procurement and Perfusion-Decellularization of Porcine Vascularized Flaps in a Customized Perfusion Bioreactor

Large volume soft tissue defects lead to functional deficits and can greatly impact the patient&#39;s quality of life. Although surgical reconstruction can be performed using autologous free flap transfer or vascularized composite allotransplantation (VCA), such methods also have disadvantages. Issues such as donor site morbidity and tissue availability limit autologous free flap transfer, while immunosuppression is a significant limitation of VCA. Engineered tissues in reconstructive surgery using decellularization/recellularization methods represent a possible solution. Decellularized tissues are generated using methods that remove native cellular material while preserving the underlying extracellular matrix (ECM) microarchitecture. These acellular scaffolds can then be subsequently recellularized with recipient-specific cells.
This protocol details the procurement and decellularization methods used to achieve acellular scaffolds in a pig model. In addition, it also provides a description of the perfusion bioreactor design and setup. The flaps include the porcine omentum, tensor fascia lata, and the radial forearm. Decellularization is performed via ex vivo perfusion of low concentration sodium dodecyl sulfate (SDS) detergent followed by DNase enzyme treatment and peracetic acid sterilization in a customized perfusion bioreactor.
Successful tissue decellularization is characterized by a white-opaque appearance of flaps macroscopically. Acellular flaps show the absence of nuclei on histological staining and a significant reduction in DNA content. This protocol can be used efficiently to generate decellularized soft tissue scaffolds with preserved ECM and vascular microarchitecture. Such scaffolds can be used in subsequent recellularization studies and have the potential for clinical translation in reconstructive surgery.

The protocol describes the surgical procurement and subsequent decellularization of vascularized porcine flaps by the perfusion of sodium dodecyl sulfate detergent through the flap vasculature in a customized perfusion bioreactor.

在定制灌注生物反应器中获取和灌注-脱细胞猪血管化皮瓣

Large volume soft tissue defects lead to functional deficits and can greatly impact the patient&#39;s quality of life. Although surgical reconstruction ...

Extraction and Dissection of the Domesticated Pig Brain

Use of the pig as a preclinical and translatable animal model has been well-documented and accepted by research fields investigating cardiovascular systems, gastrointestinal systems, and nutrition, and the pig is increasingly being used as a large animal model in neuroscience. Furthermore, the pig is an accepted model to study neurodevelopment as it displays brain growth and development patterns similar to what occurs in humans. As a less common animal model in neuroscience, surgical and dissection procedures on pigs may not be as familiar or well-practiced among researchers. Therefore, a standardized visual protocol detailing consistent extraction and dissection methods may prove valuable for researchers working with the pig. The following video showcases a technique to remove the pig brain while keeping the cortex and brainstem intact and reviews methods to dissect several commonly investigated brain regions including the brainstem, cerebellum, midbrain, hippocampus, striatum, thalamus, and medial prefrontal cortex. The purpose of this video is to provide researchers with the tools and knowledge necessary to consistently perform a brain extraction and dissection on the four-week-old pig.

This protocol details the technique for removal of the pig brain in its entirety and dissection of several brain regions commonly studied in neuroscience.

驯养猪脑的提取和解剖

Use of the pig as a preclinical and translatable animal model has been well-documented and accepted by research fields investigating cardiovascular ...

神经科学

Intubation, Central Venous Catheter, and Arterial Line Placement in Swine for Translational Research in Abdominal Transplantation Surgery

Translational surgical research models in swine are crucial for developing safe preclinical protocols. However, the success of the experimental surgeries does not solely rely on the research team's surgical skills; perioperative care and management procedures, like intubation, central venous line, and arterial line placement, are necessary and of the utmost importance for favorable experiment results. As it is uncommon for research teams to have anesthesiologists or any other staff other than the surgical team, the surgical team involved in translational research must acquire and/or develop the skills to perform the perioperative care. The purpose of this paper is to show the techniques of intubation, central venous catheter, and arterial line placement used and perfected at the Toronto Organ Preservation Laboratory over the last 10 years, to be used as a reference for future researchers joining either this team or any other lab performing translational research protocols in swine and/or abdominal transplantation.

To obtain the best possible results, surgeons performing translational research experiments should be proficient at intubation and vascular line placement. This paper describes the techniques used by the Toronto Organ Preservation Laboratory to perform these procedures.

Translational surgical research models in swine are crucial for developing safe preclinical protocols. However, the success of the experimental surgeries ...

猪生物医学模型生物库组织样品的取样策略与处理

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一个耦合试验，有限元建模方法以评估生物材料软的高应变率力学响应

活生物库的创建和维护 - 我们如何做到这一点

转化医学生物样本库:优化样品管理的标准操作程序

通过改进采样技术和组织学表征提高前列腺肿瘤生物样本库的可靠性

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Intubation, Central Venous Catheter, and Arterial Line Placement in Swine for Translational Research in Abdominal Transplantation Surgery