Name: an ex vivo choroid sprouting assay of ocular microvascular angiogenesis
Uploaded: 2020-08-06T16:30:00.000+00:00
Duration: 6 min 10 s
Description: Watch this Scientific Journal Video about An Ex Vivo Choroid Sprouting Assay of Ocular Microvascular Angiogenesis at JoVE.com

这项工作得到了曼佩铃木糖尿病基金会（YT）、波士顿儿童医院 OFD/BTREC/CTREC 教师职业发展补助金、波士顿儿童医院眼科基金会、BCH 试点奖、BCH 曼顿中心奖学金和小长颈鹿基金会 （ZF）、德国研究基金会 （DFG; 到 BC [CA1940/1-1]）， NIH R24EY024868， EY017017， R01717-13S1， EY030904-01， BCH IDDRC （1U54HD090255）， 马萨诸塞州狮子眼基金会 （LEHS）.

所述的所有动物实验都得到波士顿儿童医院机构动物护理和使用委员会的批准（ARCH协议号19-04-3913R）。
1. 准备
<ol><li>加入5 mL的青霉素/链霉素（10000 U/mL）和5 mL和10 mL的商用补充剂到500 mL的完整经典介质与血清。最初为 50 mL 的介质的 Aliquot。 注:请勿将任何介质退回库存，以免受到污染。</li>
	<li>在冰上放一完全经典介质。</li>
	<li>使用 70% 乙醇清洁解剖显微镜、钳子和剪刀。</li>
	<li>准备两个细胞培养皿（10厘米），一个在解剖显微镜上，一个在冰上;在每道菜中放入 10 mL 的完整经典介质。</li>
</ol>2. 实验步骤 （图1）
<ol><li>牺牲C57BL/6J小鼠周围的产后（P） 20使用75-100毫克/千克氯胺酮和7.5-10毫克/千克木氨酸注射在陷阱酮。解剖前，在冰上保持眼睛完全的经典介质。</li>
	<li>去除结缔组织（肌肉和脂肪组织）和眼睛上的视神经。</li>
	<li>使用微剪刀圆圆切 0.5 mm 后角膜边缘。去除角膜/虹膜复合物、玻璃和透镜。</li>
	<li>使切削边缘垂直于视神经的切口为 1 mm，并切割 1 mm 宽度的圆周带。将建筑群的中央和外围区域分开。使用钳子从RPE/胆状/四分层复合物中剥离视网膜。</li>
	<li>保持外设胆小球带在冰上完全经典介质;隔离另一只眼睛，并重复这个过程，以削减第二个带。</li>
	<li>将圆形带切成 6 = 相等的方形片（±1 mm x 1 mm）。 注:切勿接触任何边缘。</li>
	<li>根据制造商的指示解冻基底膜提取物 （BME）。将 30 μL/μL/井的 BME 添加到 24 井组织培养板的每个井的中心。确保 BME 的液滴在板底部形成凸形圆顶，而不接触边缘。 注:在冰箱中通宵解冻 BME。解冻后，BME 应随时在冰上。</li>
	<li>将组织放在 BME 的中间。 注:不要压平胆状外植;一般来说，让组织在BME内膨胀。组织的方向（侧向上或向下）不会影响实验结果。</li>
	<li>在37°C下孵育板10分钟，让凝胶凝固。</li>
	<li>加入 500 μL 的完整经典介质/井。</li>
	<li>每隔一天更换一次经典介质（500 μL）。用显微镜观察3天后，可以观察到胆状芽。 注:对于生长因子治疗，使组织挨饿4小时。稀释生长因子降低介质中的试验化合物（1:200 提升而不是 1:50）。</li>
</ol>3.SWIFT-Choroid计算机化定量方法17 （图2）
注:使用计算机化方法测量生长容器覆盖的区域。在量化之前，需要 ImageJ 软件的宏插件（有关更多详细信息 ，请参阅 补充信息）。
<ol><li>使用 ImageJ 打开 choroid 发芽图像并检查"图像|类型|8 位 "与灰度。</li>
	<li>转到"图像|调整|亮度/对比度（Ctrl/shift/C）"并优化对比度。</li>
	<li>使用魔杖功能从图像中轮廓和去除存在于芽的中心的胆状组织（使用快捷键"F1"）（图2A，B）。 注:将魔杖的容差率设置为 20-30%。</li>
	<li>使用自由选择工具移除图像的背景（图 2C）。转到"图像|调整|阈值 （Ctrl/shift/T）"。使用阈值函数在背景和外围定义微血管芽（图 2D）。</li>
	<li>单击"F2"，将显示摘要。单击"保存"来保存所选区域的图像。保存与原始图像相同的文件夹中，供将来参考。</li>
	<li>测量一组样本后，复制记录以进行数据分析。 注:也可以通过"分析"测量面积（μm2）|使用带比例条的图像设置比例"。</li>
</ol>

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提交人没有财务披露。计算机化方法通过作者免费提供给学术机构。

胆瘤发芽分析辅助研究的新血管AMD9，10，18，19，20。胆龙外植可以从老鼠以及大鼠和人类17，21分离。胆道外植包括ECs，巨噬细胞，和心状体17。在此测定胆道DC与相邻细胞（如RPE细胞）之间的相互作用，有助于阐?...

胆血管生成失调与新血管年龄相关的黄斑变性（AMD）1。胆小球是视网膜色素上皮（RPE）下的微血管床。研究表明，胆瘤中血流量的减少与AMD2的进展有关。血管内皮、RPE、巨噬细胞、心肺和其他细胞之间的复杂关系是组织3、4、5的平衡。因此，可重复的测定建模对新血管AMD的研究至关重要。
前活体血管生成测定和体外内皮细胞培养可以补充对体内微血管行为的研究，用于测试新药和机制学研究。内皮细胞，如人类视网膜微血管内皮细胞（HRMEC），人类脐带静脉内皮细胞（HUVEC），分离的原兽脑或视网膜ECs，常用于体外研究，用于眼血管生成研究6，7，8。特别是HRMEC已被广泛用作体外胆状新血管化（CNV）9的模型，通过评估内皮增殖、迁移、管状形成和血管渗漏来评估干预6、10。然而，由于在胆道中与其他细胞类型缺乏相互作用，并且由于这些测定中使用的大多数EC并非源自胆瘤，培养中的EC作为CNV的模型受到限制。在培养中，小鼠巧克力球状电子能难以分离和维护。
大动脉环测定被广泛用作大血管增殖的模型。来自大动脉外植的血管芽包括 ES、周食和巨噬细胞11。大音环测定模型大血管发生井12，13，14。然而，它作为胆囊新血管化的模型有局限性，因为大动脉环是缺乏特征的胆囊微血管环境的大血管组织，而来自大型血管的芽可能与涉及微血管病理学的毛细管网络的芽不同。最近，一个小组发表了一个前维维视网膜测定15，16。虽然，它适用于视网膜新血管疾病，但它不像在AMD中所看到的那样适合胆状新血管化。
利用小鼠RPE、胆球和胆碱外植组织进行胆瘤发芽测定，以更好地模拟CNV。组织可以很容易地从小鼠（或其他物种）的眼睛17分离。这种测定允许重复评估药理化合物的亲血管原位和抗血管原位，并评估特定途径在胆道新血管化中的作用，利用转基因小鼠和对控组18。这种胆瘤发芽测定已在许多后续出版物9，10，18，19，20被引用。在这里，演示了使用这种测定的方法。

<table><tbody><tr><td>AnaSed (Xylazine)</td><td>AKORN</td><td>59339-110-20</td><td></td></tr><tr><td>Basal membrane extract (BME) Matrigel</td><td>BD Biosciences</td><td>354230</td><td></td></tr><tr><td>Cell culture dish</td><td>NEST</td><td>704001</td><td>10cm</td></tr><tr><td>Complete classic medium with serum and CultureBoost</td><td>Cell systems</td><td>4Z0-500</td><td></td></tr><tr><td>Ethyl alcohol 200 Proof</td><td>Pharmco</td><td>111000200</td><td>use for 70%</td></tr><tr><td>Kimwipes</td><td>Kimberly-Clark</td><td>06-666</td><td></td></tr><tr><td>Microscope</td><td>ZEISS</td><td>Axio Observer Z1</td><td></td></tr><tr><td>Penicillin/Streptomycin</td><td>GIBCO</td><td>15140</td><td>10000 U/mL</td></tr><tr><td>Tissue culture plate (24-well)</td><td>Olympus</td><td>25-107</td><td></td></tr><tr><td>VetaKet CIII (Ketamine)</td><td>AKORN</td><td>59399-114-10</td><td></td></tr></tbody></table>

an ex vivo choroid sprouting assay of ocular microvascular angiogenesis

病理胆瘤血管生成，年龄相关的黄斑变性的显著特征，导致视力障碍和失明。使用人类视网膜微血管内皮细胞（HRMECs）或分离原发性视网膜EC进行内皮细胞增殖检测，在体外模型中广泛使用，用于研究视网膜血管生成。然而，分离纯杂音视网膜内皮细胞在技术上具有挑战性，视网膜ECs可能有不同的增殖反应比胆状内皮细胞和不同的细胞/细胞相互作用。开发了一种高度可重复的外体胆瘤发芽测定，作为胆球微血管增殖的模型。该模型包括胆道血管（EC、巨噬细胞、心状物）和视网膜色素上皮（RPE）之间的相互作用。小鼠RPE/胆状/细胞外植在生长因子减少基膜提取物（BME）（第0天）中分离和孵育。中等每隔一天改变一次，胆瘤发芽在第6天被量化。使用倒置相显微镜拍摄单个胆瘤外植的图像，并使用本实验室开发的 ImageJ 软件半自动宏插件对发芽区域进行量化。这种可重复的外体胆瘤发芽测定可用于评估化合物的潜在治疗和微血管疾病研究，以评估使用野生型和转基因小鼠组织参与胆小血管增殖的途径。

每天胆小球发芽生长的比较
我们解剖了带sclera的胆小球，嵌入到BME中，并培养了6天（图1）。第3天至第6天C57BL/6J小鼠的胆瘤发芽用显微镜检查，用SWIFT-Choroid对图像J中的半自动定量方法进行量化。在一个代表性的案例中，第3天（图3A）的胆状发芽区（从外植延伸的容器，不包括外植本身）为0.38毫米2（图3A），?...

Watch this Scientific Journal Video about An Ex Vivo Choroid Sprouting Assay of Ocular Microvascular Angiogenesis at JoVE.com

An Ex Vivo Choroid Sprouting Assay of Ocular Microvascular Angiogenesis

该协议提供胆瘤发芽测定，微血管增殖的外体模型。这种测定可用于评估增殖胆小血管所涉及的途径，并评估使用野生型和转基因小鼠组织的药物治疗。

眼部微血管血管生成前维沃胆球状芽分析

Pathological choroidal angiogenesis, a salient feature of age-related macular degeneration, leads to vision impairment and blindness. Endothelial cell ...

an-ex-vivo-choroid-sprouting-assay-ocular-microvascular

Research

JoVE Journal

Biology

5.5K 次观看.  Boston Children's Hospital, Harvard Medical School. 该协议提供胆瘤发芽测定，微血管增殖的外体模型。这种测定可用于评估增殖胆小血管所涉及的途径，并评估使用野生型和转基因小鼠组织的药物治疗。

视频: An Ex Vivo Choroid Sprouting Assay of Ocular Microvascular Angiogenesis

A Mouse Model of the Cornea Pocket Assay for Angiogenesis Study

A normal cornea is clear of vascular tissues. However, blood vessels can be induced to grow and survive in the cornea when potent angiogenic factors are administered 1. This uniqueness has made the cornea pocket assay one of the most used models for angiogenesis studies. The cornea composes multiple layers of cells. It is therefore possible to embed a pellet containing the angiogenic factor of interest in the cornea to investigate its angiogenic effect 2,3. Here, we provide a step by step demonstration of how to (I) produce the angiogenic factor-containing pellet (II) embed the pellet into the cornea (III) analyze the angiogenesis induced by the angiogenic factor of interest. Since the basic fibroblast growth factor (bFGF) is known as one of the most potent angiogenic factors 4, it is used here to induce angiogenesis in the cornea.

The cornea is unique in that it lacks vascular tissues. However, robust blood vessel growth and survival can be induced in the cornea by potent angiogenic factors. Therefore, the cornea can provide with us a valuable tool for angiogenic studies. This protocol demonstrates how to perform the mouse model of cornea pocket assay and how to assess the angiogenesis induced by angiogenic factors using this model.

一个小鼠模型角膜血管生成研究掌上含量

A normal cornea is clear of vascular tissues. However, blood vessels can be induced to grow and survive in the cornea when potent angiogenic factors are ...

科研

医学

An Ex Vivo Method for Time-Lapse Imaging of Cultured Rat Mesenteric Microvascular Networks

Angiogenesis, defined as the growth of new blood vessels from pre-existing vessels, involves endothelial cells, pericytes, smooth muscle cells, immune cells, and the coordination with lymphatic vessels and nerves. The multi-cell, multi-system interactions necessitate the investigation of angiogenesis in a physiologically relevant environment. Thus, while the use of in vitro cell-culture models have provided mechanistic insights, a common critique is that they do not recapitulate the complexity associated with a microvascular network. The objective of this protocol is to demonstrate the ability to make time-lapse comparisons of intact microvascular networks before and after angiogenesis stimulation in cultured rat mesentery tissues. Cultured tissues contain microvascular networks that maintain their hierarchy. Immunohistochemical labeling confirms the presence of endothelial cells, smooth muscle cells, pericytes, blood vessels and lymphatic vessels. In addition, labeling tissues with BSI-lectin enables time-lapse comparison of local network regions before and after serum or growth factor stimulation characterized by increased capillary sprouting and vessel density. In comparison to common cell culture models, this method provides a tool for endothelial cell lineage studies and tissue specific angiogenic drug evaluation in physiologically relevant microvascular networks.

An Ex Vivo Method for Time-Lapse Imaging of Cultured Rat Mesenteric Microvascular Networks

Angiogenesis involves multi-cell, multi-system interactions that need to be investigated in a physiologically relevant environment. The objective of this study is to demonstrate the ability of the rat mesentery culture model to make time-lapse comparisons of intact microvascular networks during angiogenesis.

一个<em&gt;离体</em&gt;方法培养大鼠肠系膜微血管网络的时间推移成像

Angiogenesis, defined as the growth of new blood vessels from pre-existing vessels, involves endothelial cells, pericytes, smooth muscle cells, immune ...

发育生物学

Whole Mount Immunofluorescent Staining of the Neonatal Mouse Retina to Investigate Angiogenesis In vivo

Angiogenesis is the complex process of new blood vessel formation defined by the sprouting of new blood vessels from a pre-existing vessel network. Angiogenesis plays a key role not only in normal development of organs and tissues, but also in many diseases in which blood vessel formation is dysregulated, such as cancer, blindness and ischemic diseases. In adult life, blood vessels are generally quiescent so angiogenesis is an important target for novel drug development to try and regulate new vessel formation specifically in disease. In order to better understand angiogenesis and to develop appropriate strategies to regulate it, models are required that accurately reflect the different biological steps that are involved. The mouse neonatal retina provides an excellent model of angiogenesis because arteries, veins and capillaries develop to form a vascular plexus during the first week after birth. This model also has the advantage of having a two-dimensional (2D) structure making analysis straightforward compared with the complex 3D anatomy of other vascular networks. By analyzing the retinal vascular plexus at different times after birth, it is possible to observe the various stages of angiogenesis under the microscope. This article demonstrates a straightforward procedure for analyzing the vasculature of a mouse retina using fluorescent staining with isolectin and vascular specific antibodies.

Whole Mount Immunofluorescent Staining of the Neonatal Mouse Retina to Investigate Angiogenesis In vivo

The neonatal murine retina provides a well characterized physiological model of angiogenesis, which permits investigations of the roles of different genes or drugs that modulate angiogenesis in an in vivo context. Immunofluorescent staining to accurately visualize the vascular plexus is pivotal to the success of these types of studies.

整装免疫荧光染色新生小鼠视网膜血管生成调查<em&gt;在体内</em

Angiogenesis  is the complex process of new blood vessel formation defined by the sprouting  of new blood vessels from a pre-existing vessel network. ...

神经科学

The Corneal Micropocket Assay: A Model of Angiogenesis in the Mouse Eye

The mouse corneal micropocket assay is a robust and quantitative in vivo assay for evaluating angiogenesis. By using standardized slow-release pellets containing specific growth factors that trigger blood vessel growth throughout the naturally avascular cornea, angiogenesis can be measured and quantified. In this assay the angiogenic response is generated over the course of several days, depending on the type and dose of growth factor used. The induction of neovascularization is commonly triggered by either basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF). By combining these growth factors with sucralfate and hydron (poly-HEMA (poly(2-hydroxyethyl methacrylate))) and casting the mixture into pellets, they can be surgically implanted in the mouse eye. These uniform pellets slowly-release the growth factors over five or six days (bFGF or VEGF respectively) enabling sufficient angiogenic response required for vessel area quantification using a slit lamp. This assay can be used for different applications, including the evaluation of angiogenic modulator drugs or treatments as well as comparison between different genetic backgrounds affecting angiogenesis. A skilled investigator after practicing this assay can implant a pellet in less than 5 min per eye.

The protocol describes the corneal micropocket assay as developed in mice.

角膜微囊含量：血管生成在小鼠眼模型

The mouse corneal micropocket assay is a robust and quantitative in vivo assay for evaluating angiogenesis. By using standardized slow-release pellets ...

Standardized and Scalable Assay to Study Perfused 3D Angiogenic Sprouting of iPSC-derived Endothelial Cells In Vitro

Pre-clinical drug research of vascular diseases requires in vitro models of vasculature that are amendable to high-throughput screening. However, current in vitro screening models that have sufficient throughput only have limited physiological relevance, which hinders the translation of findings from in vitro to in vivo. On the other hand, microfluidic cell culture platforms have shown unparalleled physiological relevancy in vitro, but often lack the required throughput, scalability and standardization. We demonstrate a robust platform to study angiogenesis of endothelial cells derived from human induced pluripotent stem cells (iPSC-ECs) in a physiological relevant cellular microenvironment, including perfusion and gradients. The iPSC-ECs are cultured as 40 perfused 3D microvessels against a patterned collagen-1 scaffold. Upon the application of a gradient of angiogenic factors, important hallmarks of angiogenesis can be studied, including the differentiation into tip- and stalk cell and the formation of perfusable lumen. Perfusion with fluorescent tracer dyes enables the study of permeability during and after anastomosis of the angiogenic sprouts. In conclusion, this method shows the feasibility of iPSC-derived ECs in a standardized and scalable 3D angiogenic assay that combines physiological relevant culture conditions in a platform that has the required robustness and scalability to be integrated within the drug screening infrastructure.

This method describes the culture of iPSC-derived endothelial cells as 40 perfused 3D microvessels in a standardized microfluidic platform. This platform enables the study of gradient-driven angiogenic sprouting in 3D, including anastomosis and stabilization of the angiogenic sprouts in a scalable and high-throughput manner.

标准化和可扩展的测定，以研究iPSC衍生内皮细胞在Vitro的渗透3D血管生成发芽

Pre-clinical drug research of vascular diseases requires in vitro models of vasculature that are amendable to high-throughput screening. However, current ...

生物工程

Retinal Explant of the Adult Mouse Retina as an Ex Vivo Model for Studying Retinal Neurovascular Diseases

One of the challenges in retina research is studying the cross-talk between different retinal cells such as retinal neurons, glial cells, and vascular cells. Isolating, culturing, and sustaining retinal neurons in vitro have technical and biological limitations. Culturing retinal explants may overcome these limitations and offer a unique ex vivo model to study the cross-talk between various retinal cells with well-controlled biochemical parameters and independent of the vascular system. Moreover, retinal explants are an effective screening tool for studying novel pharmacological interventions in various retinal vascular and neurodegenerative diseases such as diabetic retinopathy. Here, we describe a detailed protocol for retinal explants' isolation and culture for an extended period. The manuscript also presents some of the technical problems during this procedure that may affect the desired outcomes and reproducibility of the retinal explant culture. The immunostaining of the retinal vessels, glial cells, and neurons demonstrated intact retinal capillaries and neuroglial cells after 2 weeks from the beginning of the retinal explant culture. This establishes retinal explants as a reliable tool for studying changes in the retinal vasculature and neuroglial cells under conditions that mimic retinal diseases such as diabetic retinopathy.

Retinal Explant of the Adult Mouse Retina as an Ex Vivo Model for Studying Retinal Neurovascular Diseases

This protocol presents and describes steps for the isolation, dissection, culturing, and staining of retinal explants obtained from an adult mouse. This method is beneficial as an ex vivo model for studying different retinal neurovascular diseases such as diabetic retinopathy.

成年小鼠视网膜的视网膜外植体作为研究视网膜神经血管疾病的离 体 模型

One of the challenges in retina research is studying the cross-talk between different retinal cells such as retinal neurons, glial cells, and vascular ...

使用划痕伤口迁移和球状体发芽测定法研究血管生成

Investigating Angiogenesis on a Functional and Molecular Level by Leveraging the Scratch Wound Migration Assay and the Spheroid Sprouting Assay

Angiogenesis plays a crucial role in both physiological and pathological processes within the body including tumor growth or neovascular eye disease. A detailed understanding of the underlying molecular mechanisms and reliable screening models are essential for targeting diseases effectively and developing new therapeutic options. Several in vitro assays have been developed to model angiogenesis, capitalizing on the opportunities a controlled environment provides to elucidate angiogenic drivers at a molecular level and screen for therapeutic targets.
This study presents workflows for investigating angiogenesis in vitro using human umbilical vein endothelial cells (HUVECs). We detail a scratch wound migration assay utilizing a live cell imaging system measuring endothelial cell migration in a 2D setting and the spheroid sprouting assay assessing endothelial cell sprouting in a 3D setting provided by a collagen matrix. Additionally, we outline strategies for sample preparation to enable further molecular analyses such as transcriptomics, particularly in the 3D setting, including RNA extraction as well as immunocytochemistry. Altogether, this framework offers scientists a reliable and versatile toolset to pursue their scientific inquiries in in vitro angiogenesis assays.

作者聚焦：通过检测开发中的挑战和创新研究血管生成

This study presents the two-dimensional (2D) scratch wound migration assay and the three-dimensional (3D) spheroid sprouting assay, along with their respective downstream analysis methods, including RNA extraction and immunocytochemistry, as suitable assays to study angiogenesis in vitro.

利用划痕伤口迁移测定和球状体发芽测定在功能和分子水平上研究血管生成

Angiogenesis plays a crucial role in both physiological and pathological processes within the body including tumor growth or neovascular eye disease. A ...

In Vitro Three-Dimensional Sprouting Assay of Angiogenesis Using Mouse Embryonic Stem Cells for Vascular Disease Modeling and Drug Testing

Recent advances in induced pluripotent stem cells (iPSC) and gene editing technologies enable the development of novel human cell-based disease models for phenotypic drug discovery (PDD) programs. Although these novel devices could predict the safety and efficacy of investigational drugs in humans more accurately, their development to the clinic still strongly rely on mammalian data, notably the use of mouse disease models. In parallel to human organoid or organ-on-chip disease models, the development of relevant in vitro mouse models is therefore an unmet need for evaluating direct drug efficacy and safety comparisons between species and in vivo and in vitro conditions. Here, a vascular sprouting assay that utilizes mouse embryonic stem cells differentiated into embryoid bodies (EBs) is described. Vascularized EBs cultured onto 3D-collagen gel develop new blood vessels that expand, a process called sprouting angiogenesis. This model recapitulates key features of in vivo sprouting angiogenesis-formation of blood vessels from a pre-existing vascular network-including endothelial tip cell selection, endothelial cell migration and proliferation, cell guidance, tube formation, and mural cell recruitment. It is amenable to screening for drugs and genes modulating angiogenesis and shows similarities with recently described three-dimensional (3D) vascular assays based on human iPSC technologies.

In Vitro Three-Dimensional Sprouting Assay of Angiogenesis Using Mouse Embryonic Stem Cells for Vascular Disease Modeling and Drug Testing

This assay utilizes mouse embryonic stem cells differentiated into embryoid bodies cultured in 3D-collagen gel to analyze the biological processes that control sprouting angiogenesis in vitro. The technique can be applied for testing drugs, modeling diseases, and for studying specific genes in the context of deletions that are embryonically lethal.

体外 使用小鼠胚胎干细胞进行血管疾病建模和药物测试的血管生成的三维发芽测定

Recent advances in induced pluripotent stem cells (iPSC) and gene editing technologies enable the development of novel human cell-based disease models for ...

眼部微血管血管生成前维沃胆球状芽分析

摘要

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一个小鼠模型角膜血管生成研究掌上含量

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整装免疫荧光染色新生小鼠视网膜血管生成调查

角膜微囊含量：血管生成在小鼠眼模型

标准化和可扩展的测定，以研究iPSC衍生内皮细胞在Vitro的渗透3D血管生成发芽

成年小鼠视网膜的视网膜外植体作为研究视网膜神经血管疾病的离体模型

利用划痕伤口迁移测定和球状体发芽测定在功能和分子水平上研究血管生成

利用划痕伤口迁移测定和球状体发芽测定在功能和分子水平上研究血管生成

利用划痕伤口迁移测定和球状体发芽测定在功能和分子水平上研究血管生成

体外使用小鼠胚胎干细胞进行血管疾病建模和药物测试的血管生成的三维发芽测定

眼部微血管血管生成前维沃胆球状芽分析

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一个小鼠模型角膜血管生成研究掌上含量

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整装免疫荧光染色新生小鼠视网膜血管生成调查

角膜微囊含量：血管生成在小鼠眼模型

标准化和可扩展的测定，以研究iPSC衍生内皮细胞在Vitro的渗透3D血管生成发芽

成年小鼠视网膜的视网膜外植体作为研究视网膜神经血管疾病的离 体 模型

利用划痕伤口迁移测定和球状体发芽测定在功能和分子水平上研究血管生成

利用划痕伤口迁移测定和球状体发芽测定在功能和分子水平上研究血管生成

利用划痕伤口迁移测定和球状体发芽测定在功能和分子水平上研究血管生成

体外 使用小鼠胚胎干细胞进行血管疾病建模和药物测试的血管生成的三维发芽测定

成年小鼠视网膜的视网膜外植体作为研究视网膜神经血管疾病的离体模型

体外使用小鼠胚胎干细胞进行血管疾病建模和药物测试的血管生成的三维发芽测定