Name: the in ovo chick chorioallantoic membrane (cam) assay as an efficient xenograft model of hepatocellular carcinoma
Uploaded: 2015-10-09T14:00:00.000+00:00
Duration: 8 min 17 s
Description: Watch this Scientific Journal Video about The In Ovo Chick Chorioallantoic Membrane CAM Assay as an Efficient Xenograft Model of Hepatocellular Carcinoma at JoVE.com

A.S. was partially supported by grants from the National Institutes of Health (NIH) National Institute of Dental and Craniofacial Research (5R03DE021741-02) and the National Cancer Institute (1K08CA154963-01A1).&#160;
The Howard Hughes Medical Institute provided funding for M.L. through the HHMI Medical Research Fellows Program.

1.卵孵化<ol><li>获得8天龄的无特定病原体鸡胚。 </li><li>将鸡蛋在旋转蛋托，朝上加盖结束，并把一个鸡蛋孵化器内的旋转托盘。孵化鸡蛋48小时在36℃和50％的湿度。 </li></ol> 2.删除的CAM和打开鸡蛋<ol><li>从孵化器收集的鸡蛋。将鸡蛋放在蛋架，加盖的一面朝上，并提请层流罩。 </li><li>关闭室和发动机罩灯。 </li><li>轻轻握住蛋的加盖端蛋坎德勒以暴露所述CAM的脉管以及空气囊。 注:蛋烛台是用于可视化发育​​中的胚胎和相关的空气囊和血管的光源。 </li><li>在两个主要血管之间的铅笔标记。 </li><li>开灯，并使用无菌推针使一个孔在气囊（加盖端）之上蛋的尖端和一个孔在所述的pencil大关。不要推则推销一路过关斩将;大约3毫米的孔深通常就足够了。 </li><li>把灯关掉。挤压安全球，稳稳地按下开口端对上方的空气囊孔。应用吸拉空气进入孔在铅笔标记，导致CAM下落离开壳的铅笔标记。 </li><li>检查空气囊已经从卵到使用坎德勒铅笔标记孔加盖结束感动。如果没有，用图钉，使两个孔略深，然后用安全球重新申请吸力。 </li><li>开灯。拿着一手拿鸡蛋，打开DREMEL旋转工具连接了15/16英寸的切割轮，并两次横向切割只是深足通过壳削减，但不够深削减一路下跌至郁闷CAM。使约2厘米长，1厘米彼此间隔开的切口。使切口足够宽，从而硅环1厘米，直径可通过但比标准的透明胶带宽度更薄。 </li><li>接着，通过轻轻接触切割轮与壳之间做出，并在两次横向切割的端部1纵向切口。 </li><li>滑动壳片和平行于下壳无菌镊子。抢壳片的镊子和删除干净整片。 </li><li>将透明胶带在新打开的窗口中，确保了自身折叠一端，便于拆卸。 </li><li>将鸡蛋放回孵化器中的蛋托（而不是在旋转蛋架），直到接种时间。 </li></ol> 3.接种<ol><li>保持基底膜蛋白混合物，如基质胶，在冰上（防止聚合），并装在罩。 </li><li>使用2mM EDTA的分离的细胞（4 - 每个T75烧瓶5毫升）并置于烧瓶中的组织培养培养箱中5分钟。 </li><li>重悬分离的细胞介质，并使用血球和台盼蓝计数。间50万吨使用Ø2000000细胞移植到CAM，取决于细胞系。优化在初步实验中的适当数量。 </li><li>在500 XG需要合适的细胞数量和离心5分钟测量体积。 </li><li>弃上清，然后悬浮细胞沉淀在40μ微升PBS ++和20μ升基底膜的混合物。 注意:在这一点上，细胞可以用药物或其他测试试剂如重悬到细胞/基底膜混合物悬浮所需实验处理。另外，代理可以嫁接后吸至电池/基底膜混合物悬浮体。 </li><li>检索与从培养箱和发生在层流罩鸡蛋蛋架。 </li><li>剥离胶带关闭窗口，检索从使用灭菌镊子低温小瓶的帽的1毫米硅胶圈，并通过窗口滴环。 </li><li>吸取PBS的60μ升细胞溶液++和基底膜mixturË直接进入硅胶圈搁在所述CAM的中心。 </li><li>胶带重新贴到窗口，移动蛋架回孵化器。在这一点上，请小心地移动鸡蛋为环和细胞悬浮液时，很容易成为流离失所。 </li><li>允许肿瘤生长5至7天。 </li></ol> 4.收获肿瘤<ol><li>在引擎盖下面垫的地方。根据需要，用于组织学准备35×10mm的板和多聚甲醛的容器。放1毫升的PBS ++中的每一个35×10毫米的钢板。 </li><li>将生物危害袋罩。以无菌解剖剪刀和在接近年​​底加盖孔插入尖端进入卵子。 </li><li>整个鸡蛋围绕纵向切开。 注意:所述CAM和肿瘤与环应粘到外壳与窗口的上半部分。 </li><li>用解剖剪和钳子，解剖肿瘤远离CAM和放置肿瘤中的35×10毫米的钢板。测量和称重肿瘤。在paraformaldehy修复肿瘤德然后石蜡嵌入它们的组织学和免疫组织化学实验。 </li></ol> 5.（可选）:肿瘤细胞分离<ol><li>使用夹层剪刀剁碎在35×10毫米的钢板肿瘤。倾切碎肿瘤入15ml离心管中，并除去尽可能多的PBS ++尽可能通过移液。 </li><li>加入2毫升胶原酶（1mg / ml的在PBS ++），转管数次，孵育管中在37℃下至少30分钟（长达数小时）。 </li><li>加入5 ml的媒体到每个管中，通过上下抽吸20重悬彻底 - 40倍。彻底移液是完整的细胞分离的关键。 </li><li>允许沉淀沉降，将上清液转移到新的管中。离心在500×g离心5分钟。除去上清液，留下游离的细胞团。 注意:在这一点上，细胞沉淀可用于各种实验（流式细胞术，裂解为在Western印迹，RNA分离等用途）。总肿瘤细胞计数也可使用血细胞计数器和台盼蓝染色获得的。需要注意的是较小的，椭圆形的细胞是鸡细胞从CAM和不应该算作肿瘤细胞。应该有污染性非肿瘤（CAM）细胞如果肿瘤是从所述CAM解剖远仔细的最少数量。 </li></ol>

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A.S.&#160;receives&#160;funding from the Advaxis Pharmaceutical company to support an investigator-initiated clinical trial.&#160;

在这个协议中几个关键步骤最有可能占了改进的胚胎存活以及肿瘤生长的增加的可靠性。通过施加抽吸到空气囊丢弃所述CAM远离壳是侵入性比其它现有的方法（使用针从卵，钝性分离等除去白蛋白）以下。使用无菌图钉创造这种方法需要使用两个小孔是协议中最具挑战性的部分。使用过大的力可能会损坏到CAM及其脉管通过过度穿透的推针或者甚至可能导致破裂或破坏的蛋。使用图钉动手实践，使?...

肝细胞癌（HCC）是癌症死亡率在世界1中的第三大原因。目前，只有30％的肝癌患者有资格有可能治愈手术治疗2，和全身化疗不灵验3。因此，有迫切未满足的临床需要新的肝癌的治疗，并且适用于新的代理程序有效的测试模型系统的发展。鸡胚绒毛尿囊膜（CAM）试验提供了在体内测试潜在的抗肿瘤药物的可重复的，具有成本效益，并快速介质通量方法。 CAM分析已被广泛用于研究血管生成4。还已经成功地发展成癌症的肿瘤异种移植物模型，包括成胶质细胞瘤5，胰腺癌6，黑素瘤7-9和骨肉瘤10-11 无论是在OVO 12 和前OVO13技术已被用在文献中，以改变从协议到协议的细节。一个主要的挑战CAM移植瘤模型是胚胎死亡的操控卵子后的发病率比较高，与已发表 ​​的鸡胚死亡率从25 - 50％的11-14。 在这篇文章中，我们描述了一个卵内肝癌移植瘤模型，可靠地产生三维，血管瘤的病理类似未分化肝癌生长的发展。我们已经适应首先由Ossowski 等人 14所述的协议，并已实现了高达93％鸡胚存活率具有极高的肿瘤植入。

<table><tbody><tr><td>Premium incubated eggs</td><td>Charles River</td><td>N/A</td><td>http://www.criver.com/files/pdfs/avian/av_c_spf_egg_price_list.aspx</td></tr><tr><td>Egg incubators</td><td>GQF</td><td>Hova-Bator 2362N</td><td></td></tr><tr><td>Rotating egg trays</td><td>GQF</td><td>1611 Automatic Egg Turner</td><td></td></tr><tr><td>Egg candler</td><td>Lyon</td><td>Hi-Power 950-070</td><td></td></tr><tr><td>Dremel 100 rotary tool with 15/16 cut-off wheel</td><td>Dremel</td><td>100-N/7</td><td></td></tr><tr><td>Sterile forceps, push pin, dissection scissors, Scotch tape</td><td></td><td></td><td></td></tr><tr><td>Matrigel</td><td>BD Biosciences</td><td>356234</td><td></td></tr><tr><td>Cryogenic vials, external thread with silicone washer</td><td>Corning</td><td>430659</td><td></td></tr><tr><td>Collagenase from Clostridium histolyticum</td><td>Sigma</td><td>C9891</td><td></td></tr></tbody></table>

the in ovo chick chorioallantoic membrane (cam) assay as an efficient xenograft model of hepatocellular carcinoma

鸡胚绒毛尿囊膜（CAM）开始7天受精后制定和成熟通过12天的CAM是自然免疫缺陷和高度血管，使其成为肿瘤植入的理想系统。此外，所述CAM包含细胞外基质蛋白，如纤连蛋白，层粘连蛋白，胶原蛋白，整合素（v）的β3，和MMP-2，使其成为有吸引力的模型来研究肿瘤侵袭和转移。科学家长期以来采取所述CAM的生理学优点通过使用它作为血管生成的模型。最近，CAM分析已被修改，使其成为对各种癌症的体内异种移植物模型体系桥接基本体外工作和更复杂的动物癌症模型之间的差距。 CAM分析允许肿瘤生长，抗肿瘤疗法，和亲肿瘤分子途径以生物相关的系统，既具有成本和时间有效的研究。在这里，我们描述了CAM的Xen的发展肝细胞癌（HCC）的高达93％的胚胎存活率和可靠的肿瘤ograft模型上导致立体，血管瘤的生长。

在协议的关键步骤代表性照片在这里显示。 图1A示范了使用坎德勒形象化发育中的胚胎中，空气囊，和所述CAM的脉管系统的图1B-1C示出通过使丢弃的CAM的过程的两个孔，然后施加负压使用安全灯泡，并且图1D示出一个成功删除的CAM用铅笔标记孔下一个大的气泡。 图1E和图1F展示DREMEL旋转工具的使用，以进行必要的削减在上面的气泡，并...

Watch this Scientific Journal Video about The In Ovo Chick Chorioallantoic Membrane CAM Assay as an Efficient Xenograft Model of Hepatocellular Carcinoma at JoVE.com

The In Ovo Chick Chorioallantoic Membrane CAM Assay as an Efficient Xenograft Model of Hepatocellular Carcinoma

The chick chorioallantoic membrane (CAM) is immunodeficient and highly vascularized, making it a natural in vivo model of tumor growth and angiogenesis. In this protocol, we describe a reliable method of growing three-dimensional, vascularized hepatocellular carcinoma (HCC) tumors using the CAM assay.

该<em&gt;卵内</em&gt;鸡胚尿囊膜（CAM）含量为肝癌的有效异种移植模型

The chick chorioallantoic membrane (CAM) begins to develop by day 7 after fertilization and matures by day 12. The CAM is naturally immunodeficient and ...

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Research

JoVE Journal

Medicine

The In Ovo Chick Chorioallantoic Membrane (CAM) Assay as an Efficient Xenograft Model of Hepatocellular Carcinoma

35.2K 次观看.  Icahn School of Medicine at Mount Sinai. The chick chorioallantoic membrane (CAM) is immunodeficient and highly vascularized, making it a natural in vivo model of tumor growth and angiogenesis. In this protocol, we describe a reliable method of growing three-dimensional, vascularized hepatocellular carcinoma (HCC) tumors using the CAM assay.

视频: The In Ovo Chick Chorioallantoic Membrane CAM Assay as an Efficient Xenograft Model of Hepatocellular Carcinoma

Using the Chicken Chorioallantoic Membrane In Vivo Model to Study Gynecological and Urological Cancers

Mouse models are the benchmark tests for in vivo cancer studies. However, cost, time, and ethical considerations have led to calls for alternative in vivo cancer models. The chicken chorioallantoic membrane (CAM) model provides an inexpensive, rapid alternative that permits direct visualization of tumor development and is suitable for in vivo imaging. As such, we sought to develop an optimized protocol for engrafting gynecological and urological tumors into this model, which we present here. Approximately 7 days postfertilization, the air cell is moved to the vascularized side of the egg, where an opening is created in the shell. Tumors from murine and human cell lines and primary tissues can then be engrafted. These are typically seeded in a mixture of extracellular matrix and medium to avoid cellular dispersal and provide nutrient support until the cells recruit a vascular supply. Tumors may then grow for up to an additional 14 days prior to the eggs hatching. By implanting cells stably transduced with firefly luciferase, bioluminescence imaging can be used for the sensitive detection of tumor growth on the membrane and cancer cell spread throughout the embryo. This model can potentially be used to study tumorigenicity, invasion, metastasis, and therapeutic effectiveness. The chicken CAM model requires significantly less time and financial resources compared to traditional murine models. Because the eggs are immunocompromised and immune tolerant, tissues from any organism can potentially be implanted without costly transgenic animals (e.g., mice) required for implantation of human tissues. However, many of the advantages of this model could potentially also be limitations, including the short tumor generation time and immunocompromised/immune tolerant status. Additionally, although all tumor types presented here engraft in the chicken chorioallantoic membrane model, they do so with varying degrees of tumor growth.

We present the chicken chorioallantoic membrane model as an alternative, transplantable, in vivo model for the engraftment of gynecological and urological cancer cell lines and patient-derived tumors.

利用活体中鸡胆膜研究妇科和泌尿癌

Mouse models are the benchmark tests for in vivo cancer studies. However, cost, time, and ethical considerations have led to calls for alternative in vivo ...

科研

癌症研究

The In ovo CAM-assay as a Xenograft Model for Sarcoma

Sarcoma is a very rare disease that is heterogeneous in nature, all hampering the development of new therapies. Sarcoma patients are ideal candidates for personalized medicine after stratification, explaining the current interest in developing a reproducible and low-cost xenotransplant model for this disease. The chick chorioallantoic membrane is a natural immunodeficient host capable of sustaining grafted tissues and cells without species-specific restrictions. In addition, it is easily accessed, manipulated and imaged using optical and fluorescence stereomicroscopy. Histology further allows detailed analysis of heterotypic cellular interactions.
This protocol describes in detail the in ovo grafting of the chorioallantoic membrane with fresh sarcoma-derived tumor tissues, their single cell suspensions, and permanent and transient fluorescently labeled established sarcoma cell lines (Saos-2 and SW1353). The chick survival rates are up to 75%. The model is used to study graft- (viability, Ki67 proliferation index, necrosis, infiltration) and host (fibroblast infiltration, vascular ingrowth) behavior. For localized grafting of single cell suspensions, ECM gel provides significant advantages over inert containment materials. The Ki67 proliferation index is related to the distance of the cells from the surface of the CAM and the duration of application on the CAM, the latter determining a time frame for the addition of therapeutic products.

The In ovo CAM-assay as a Xenograft Model for Sarcoma

The in ovo chorioallantoic membrane (CAM) is grafted with fresh sarcoma-derived tumor tissues, their single cell suspensions, and permanent and transient fluorescently labeled established sarcoma cell lines. The model is used to study graft- (viability, Ki67 proliferation index, necrosis, infiltration) and host (fibroblast infiltration, vascular ingrowth) behavior.

该<em&gt;在OVO</em&gt; CAM作为一个肉瘤Xenograftmodel的检测

Sarcoma is a very rare disease that is heterogeneous in nature, all hampering the development of new therapies. Sarcoma patients are ideal candidates for ...

医学

Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

In vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate the human disease provide a valuable platform for research into disease pathophysiology and for the preclinical evaluation of novel therapies. We present a variety of methods to generate subcutaneous or orthotopic human HCC xenografts in immunodeficient mice that could be utilized in a variety of research applications. With a focus on the use of primary tumor tissue from patients undergoing surgical resection as a starting point, we describe the preparation of cell suspensions or tumor fragments for xenografting. We describe specific techniques to xenograft these tissues i) subcutaneously; or ii) intrahepatically, either by direct implantation of tumor cells or fragments into the liver, or indirectly by injection of cells into the mouse spleen. We also describe the use of partial resection of the native mouse liver at the time of xenografting as a strategy to induce a state of active liver regeneration in the recipient mouse that may facilitate the intrahepatic engraftment of primary human tumor cells. The expected results of these techniques are illustrated. The protocols described have been validated using primary human HCC samples and xenografts, which typically perform less robustly than the well-established human HCC cell lines that are widely used and frequently cited in the literature. In comparison with cell lines, we discuss factors which may contribute to the relatively low chance of primary HCC engraftment in xenotransplantation models and comment on technical issues that may influence the kinetics of xenograft growth. We also suggest methods that should be applied to ensure that xenografts obtained accurately resemble parent HCC tissues.

Human tumor xenografts in immunodeficient mice are valuable tools to study cancer biology. Specific protocols to generate subcutaneous and intrahepatic xenografts from human hepatocellular carcinoma cells or tumor fragments are described. Liver regeneration induced by partial hepatectomy in recipient mice is presented as a strategy to facilitate intrahepatic engraftment.

皮下及肝内人肝癌裸鼠移植瘤的免疫缺陷小鼠的产生

In  vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate  the human disease provide a valuable platform for research into disease  ...

Chick ex ovo Culture and ex ovo CAM Assay: How it Really Works

Chicken eggs in the early phase of breeding are between in vitro and in vivo systems and provide a vascular test environment not only to study angiogenesis but also to study tumorigenesis. After the chick chorioallantoic membrane (CAM) has developed, its blood vessel network can be easily accessed, manipulated and observed and therefore provides an optimal setting for angiogenesis assays. Since the lymphoid system is not fully developed until late stages of incubation, the chick embryo serves as a naturally immunodeficient host capable of sustaining grafted tissues and cells without species-specific restrictions. In addition to nurturing developing allo- and xenografts, the CAM blood vessel network provides a uniquely supportive environment for tumor cell intravasation, dissemination, and vascular arrest and a repository where arrested cells extravasate to form micro metastatic foci.
For experimental purposes, in most of the recent studies the CAM was exposed by cutting a window through the egg shell and experiments were carried out in ovo, resulting in significant limitations in the accessibility of the CAM and possibilities for observation and photo documentation of effects. When shell-less cultures of the chick embryo were used1-4, no experimental details were provided and, if published at all, the survival rates of these cultures were low. We refined the method of ex ovo culture of chick embryos significantly by introducing a rationally controlled extrusion of the egg content. These ex ovo cultures enhance the accessibility of the CAM and chick embryo, enabling easy in vivo documentation of effects and facilitating experimental manipulation of the embryo. This allows the successful application to a large number of scientific questions: (1) As an improved angiogenesis assay5,6, (2) an experimental set up for facilitated injections in the vitreous of the chick embryo eye7-9, (3) as a test environment for dissemination and intravasation of dispersed tumor cells from established cell lines inoculated on the CAM10-12, (4) as an improved sustaining system for successful transplantation and culture of limb buds of chicken and mice13 as well as (5) for grafting, propagation, and re-grafting of solid primary tumor tissue obtained from biopsies on the surface of the CAM14.
In this video article we describe the establishment of a refined chick ex ovo culture and CAM assay with survival rates over 50%. Besides we provide a step by step demonstration of the successful application of the ex ovo culture for a large number of scientific applications. 
Daniel S. Dohle, Susanne D. Pasa, and Sebastian Gustmann contributed equally to this study.

Chick ex ovo Culture and ex ovo CAM Assay: How it Really Works

The chick chorioallantoic membrane (CAM) is a unique, naturally immunodeficient supportive culture environment to study angiogenesis and tumorigenesis. This video article demonstrates the different steps in chick ex ovo culture, application of potentially angiogenic substances and successful inoculation of tumor cells and tissues on the surface of the CAM.

小鸡前OVO文化前OVO CAM含量：它是如何真正起作用

Chicken eggs in the early phase of breeding are between in vitro and in vivo systems and provide a vascular test environment not only to study ...

生物学

A Label-free Xenograft Model for Investigating the Behavior of Human Stem Cell Spheroids in Chick Embryos

Xenografts are valuable methods to investigate the behavior of human cells in vivo. In particular, the embryonic environment provides cues for cell migration, differentiation, and morphogenesis, with unique instructive signals and germ layer identity that are often absent from adult xenograft models. In addition, embryonic models cannot discriminate self versus non-self tissues, eliminating the risk of rejection of the graft and the need for immune suppression of the host. This paper presents a methodology for transplantation of spheroids of human cells into chicken embryos, which are accessible, amenable to manipulation, and develop at 37 &#176;C. 
Spheroids allow the selection of a specific region of the embryo for transplantation. After being grafted, the cells become integrated into the host tissue, allowing the follow-up of their migration, growth, and differentiation. This model is flexible enough to allow the utilization of different adherent populations, including heterogeneous primary cell populations and cancer cells. To circumvent the need for prior cell labeling, a protocol for the identification of donor cells through hybridization of human-specific Alu probes is also described, which is particularly important when investigating heterogeneous cell populations. Furthermore, DNA probes can be easily adapted to identify other donor species. This protocol will describe the general methods for preparing spheroids, grafting into chicken embryos, fixing and processing tissue for paraffin sectioning, and finally identifying the human cells using DNA in situ hybridization. Suggested controls, examples of interpretation of results and various cell behaviors that can be assayed will be discussed in addition to the limitations of this method.

Here, we describe a method to transplant and identify human cell spheroids into chick embryos. This xenograft model uses the embryonic microenvironment as a source of instructive signals to assay cell migration, differentiation, and tropism and is especially suited for the study of primary and/or heterogeneous cell populations.

Xenografts are valuable methods to investigate the behavior of human cells in vivo. In particular, the embryonic environment provides cues for cell ...

发育生物学

The Chick Chorioallantoic Membrane In Vivo Model to Assess Perineural Invasion in Head and Neck Cancer

Perineural invasion is a phenotype in which cancer surrounds or invades the nerves. It is associated with poor clinical outcome for head and neck squamous cell carcinoma and other cancers. Mechanistic studies have shown that the molecular crosstalk between nerves and tumor cells occurs prior to physical interaction. There are only a few in vivo models to study perineural invasion, especially to investigate early progression, before physical nerve-tumor interactions occur. The chick chorioallantoic membrane model has been used to study cancer invasion, because the basement membrane of the chorionic epithelium mimics that of human epithelial tissue. Here we repurposed the chick chorioallantoic membrane model to investigate perineural invasion, grafting rat dorsal root ganglia and human head and neck squamous cell carcinoma cells onto the chorionic epithelium. We have demonstrated how this model can be useful to evaluate the ability of cancer cells to invade neural tissue in vivo.&#160;

The Chick Chorioallantoic Membrane In Vivo Model to Assess Perineural Invasion in Head and Neck Cancer

Perineural invasion is an aggressive phenotype for head and neck squamous cell carcinomas and other tumors. The chick chorioallantoic membrane model has been used for studying angiogenesis, cancer invasion, and metastasis. Here we demonstrate how this model can be utilized to assess perineural invasion in vivo.

Vivo模型中的小鸡胆膜模型,用于评估头颈部癌症的围神经入侵

Perineural invasion is a phenotype in which cancer surrounds or invades the nerves. It is associated with poor clinical outcome for head and neck squamous ...

The Chick Chorioallantoic Membrane (CAM) Model as a Tool to Study Ovarian Tissue Transplantation

Ovarian tissue cryopreservation and transplantation is an effective strategy for preserving fertility but has one major drawback, namely massive follicle loss occurring shortly after reimplantation due to abnormal follicle activation and death. Rodents are benchmark models for investigating follicle activation, but the cost, time, and ethical considerations are becoming increasingly prohibitive, thus driving the development of alternatives. The chick chorioallantoic membrane (CAM) model is particularly attractive, being inexpensive and maintaining&#160;natural immunodeficiency up to day 17 postfertilization, making it ideal to study&#160;short-term xenografting of human ovarian tissue. The CAM is also highly vascularized and has been widely used as a model to explore angiogenesis. This gives it a remarkable advantage over in vitro models and allows the investigation of mechanisms affecting the early post-grafting follicle loss process. The protocol outlined herein aims to describe the development of a CAM xenografting model for human ovarian tissue, with specific insights into the effectiveness of the technique, the graft revascularization time frame, and the tissue viability across a 6 day grafting period.

The Chick Chorioallantoic Membrane CAM Model as a Tool to Study Ovarian Tissue Transplantation

Here, we describe a protocol for developing a chick chorioallantoic membrane (CAM) xenografting model for human ovarian tissue and demonstrate the effectiveness of the technique, the graft revascularization time frame, and the tissue viability across a 6 day grafting period.

鸡绒毛膜尿囊膜 （CAM） 模型作为研究卵巢组织移植的工具

Ovarian tissue cryopreservation and transplantation is an effective strategy for preserving fertility but has one major drawback, namely massive follicle ...

Chicken Chorioallantoic Membrane-based Cancer Modeling: An In Ovo Model to Study Cancer Cell Tumorigenesis and Metastasis

Source: Sharrow, A. C. et al. <a href="https://www.jove.com/t/60651">Using the Chicken Chorioallantoic Membrane In Vivo Model to Study Gynecological and Urological Cancers.</a> J. Vis. Exp.&#160;(2020)
In this video, we describe the implantation of cancer cells onto a chicken chorioallantoic membrane to generate an in ovo model. The model successfully engrafts the ovarian cancer cells to study the progression of gynecological cancer.

基于鸡绒毛膜尿囊膜的癌症建模:研究癌细胞肿瘤发生和转移的 In-Ovo 模型

Source: Sharrow, A. C. et al. Using the Chicken Chorioallantoic Membrane In Vivo Model to Study Gynecological and Urological Cancers. J. Vis. ...

JoVE Encyclopedia of Experiments

Gynecologic Cancer

Animal model studies

Chick Chorioallantoic Membrane or CAM Tumor Model: An In Ovo Method to Simulate Perineural Invasion of Tumor Cells

Source: Schmidt, L. B. et al. <a href="https://www.jove.com/video/59296" target="_blank">The Chick Chorioallantoic Membrane In Vivo Model to Assess Perineural Invasion in Head and Neck Cancer.</a> J. Vis. Exp. (2019)
In this video, we describe the method to co-culture dorsal root ganglia and cancer cells on chick chorioallantoic membrane or CAM to study perineural invasion.

小鸡绒毛膜尿囊膜或 CAM 肿瘤模型:一种模拟肿瘤细胞神经周围浸润的 In Ovo 方法

Source: Schmidt, L. B. et al. The Chick Chorioallantoic Membrane In Vivo Model to Assess Perineural Invasion in Head and Neck Cancer. J. Vis. Exp. (2019)
...

Head and Neck Cancer

In vivo studies

Chick Chorioallantoic Membrane Harvest: A Method to Isolate Tumor Graft-bearing Chorioallantoic Membrane from Chicken Egg

Source: Schmidt, L. B. et al. <a href="https://www.jove.com/video/59296" target="_blank">The Chick Chorioallantoic Membrane In Vivo Model to Assess Perineural Invasion in Head and Neck Cancer.</a> J. Vis. Exp. (2019)
This video demonstrates the isolation of tumor-bearing chorioallantoic membrane from the chicken embryo. The harvested CAM can be used for further downstream applications.

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利用活体中鸡胆膜研究妇科和泌尿癌

该

皮下及肝内人肝癌裸鼠移植瘤的免疫缺陷小鼠的产生

小鸡前OVO文化前OVO CAM含量：它是如何真正起作用

A Label-free Xenograft Model for Investigating the Behavior of Human Stem Cell Spheroids in Chick Embryos

Vivo模型中的小鸡胆膜模型,用于评估头颈部癌症的围神经入侵

鸡绒毛膜尿囊膜（CAM）模型作为研究卵巢组织移植的工具

基于鸡绒毛膜尿囊膜的癌症建模:研究癌细胞肿瘤发生和转移的 In-Ovo 模型

小鸡绒毛膜尿囊膜或 CAM 肿瘤模型:一种模拟肿瘤细胞神经周围浸润的 In Ovo 方法

小鸡绒毛膜尿囊膜收获:一种从鸡蛋中分离携带肿瘤移植物的绒毛膜尿囊膜的方法

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摘要

探索更多视频

此视频中的章节

利用活体中鸡胆膜研究妇科和泌尿癌

该

皮下及肝内人肝癌裸鼠移植瘤的免疫缺陷小鼠的产生

小鸡前OVO文化前OVO CAM含量：它是如何真正起作用

A Label-free Xenograft Model for Investigating the Behavior of Human Stem Cell Spheroids in Chick Embryos

Vivo模型中的小鸡胆膜模型,用于评估头颈部癌症的围神经入侵

鸡绒毛膜尿囊膜 （CAM） 模型作为研究卵巢组织移植的工具

基于鸡绒毛膜尿囊膜的癌症建模:研究癌细胞肿瘤发生和转移的 In-Ovo 模型

小鸡绒毛膜尿囊膜或 CAM 肿瘤模型:一种模拟肿瘤细胞神经周围浸润的 In Ovo 方法

小鸡绒毛膜尿囊膜收获:一种从鸡蛋中分离携带肿瘤移植物的绒毛膜尿囊膜的方法

鸡绒毛膜尿囊膜（CAM）模型作为研究卵巢组织移植的工具