作者没有什么可透露的。

所有程序,包括人类受试者,均获得佛法癌症医院伦理委员会的批准,证书号为040/KEPK/VII/2017。所有患者都签署了同意书,并表示同意参与本研究。
注:纳入标准是早期乳腺癌诊断的患者,肿瘤阶段T1-T2无可触手可及和超声检查淋巴结扩大(cNo)。排除标准是局部晚期乳腺癌,接受新佐剂化疗或激素治疗,和怀孕。
1. 1%亚甲蓝染料的制备及注射技术
<ol><li>麻醉病人后,对手术场进行消毒。</li>
	<li>用3 mL注射器从小瓶中吸出2 mL的1%亚甲基蓝色染料。</li>
	<li>画一条线 , 以标记下部发线以下的胸围主要肌肉的侧界。</li>
	<li>将 2 mL 的 1% 亚甲基蓝色染料注入乳房的亚雷欧尔空间。</li>
	<li>如果乳房或 NAC 的上外象限有疤痕,则用线心针(12 MHz)在超声波指导下用 23G 针向疤痕的侧侧向腹腔内注射 2 mL 的 1% 亚甲蓝色染料。</li>
	<li>在注射部位循环按摩乳房5分钟,然后继续做手术。</li>
</ol>2. 乳房保存手术的哨兵节点活检技术(BCS)
注:手术是在接受BCS和SNB的患者身上进行的。
<ol><li>准备手术工具:单极电烧、德巴基钳子/解剖钳子和缩回器。</li>
	<li>切开皮肤、皮下组织和筋膜。</li>
	<li>找到蓝色结节或蓝色淋巴片。跟随蓝色区域,直到蓝色结点或带淋巴蓝色区域的非蓝色节点可识别。</li>
	<li>如果找不到蓝色结点或蓝色淋巴带,则沿闭周神经和侧胸静脉搜索哨点节点。</li>
	<li>仔细切除哨兵节点,避免损坏节点。</li>
	<li>将心氧化空间,以找到额外的可疑恶性淋巴结增大。</li>
</ol>3. 乳房切除术中的哨兵节点活检技术
注:手术是在接受乳房切除术和SNB的病人身上进行的。
<ol><li>切开皮肤和皮下组织。</li>
	<li>创建皮瓣。</li>
	<li>从胸腔主要去除乳房,直到可充分暴露的阴垂fosa。</li>
	<li>将锁骨筋膜切开以找到哨点节点。</li>
	<li>如果找不到蓝色淋巴带,则沿闭胆神经和侧胸静脉区域找到哨点节点。</li>
	<li>删除哨兵节点。</li>
	<li>通过触觉寻找其他可疑的淋巴结。</li>
</ol>4. 术中检查
<ol><li>切淋巴结不超过2毫米,平行于长轴。</li>
	<li>从每个节点进行触摸印记细胞学。</li>
	<li>将手术标本放在金属组织盘上,嵌入与冷冻组织密度相同的凝胶状介质中。</li>
	<li>提交所有节点进行冻结部分 (FS) 检查。</li>
	<li>将哨点节点的转移状态分类为阳性或阴性,并在手术期间向外科医生报告。</li>
</ol>5. 病理检查
<ol><li>对正式固定和石蜡嵌入组织部分的哨兵节点进行最终病理评估。</li>
	<li>根据美国癌症联合委员会(AJCC)第6版手册对哨点节点转移进行分类。巨转移(MAC)定义为大于2毫米的肿瘤沉积物,微转移(MiC)定义为0.2至2 mm之间的肿瘤沉积,分离肿瘤细胞(ITC)定义为不超过0.2毫米的细胞簇。</li>
	<li>当对ITC的定义有疑问时,对细胞角蛋白进行序列切片和免疫组化学分析。</li>
	<li>以类似的方式检查其他的阳部淋巴结。</li>
</ol>

<ol>
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我们感谢穆罕默德、M.D.阿里·阿卜杜勒·阿齐兹先生、阿迪亚·巴尤先生和阿里安达·哈里阿迪先生协助编写手稿。我们还感谢特加尔·哈里马先生在视频编辑方面提供的帮助。

在现代乳腺癌手术时代,瑞士央行已经取代ALND作为早期乳腺癌的乳腺分期护理标准,如果SN没有转移14,15,ALND应该放弃。发达国家常用的淋巴图谱技术是放射性同位素示踪剂和PBD作为组合或单一技术的应用。当无法获得放射性同位素甚至多溴联苯时,提出了如何执行瑞士国民计划的问题。我们描述的仅使用 MBD 的技...

肛门淋巴结(ALNs)转移状态是乳腺癌最重要的预后因素。Axill淋巴结解剖(ALND)是评估ALN1,2转移状态的常规程序。不幸的是,ALND导致患者的病态,降低生活质量,特别是通过增加淋巴水肿的风险后,这个程序3,4。如今 , 哨点节点活检 ( SNB ) 已经取代 ALND 为分期 , 因为它在患者中最小的疾病 5 。执行SNB的最常见方法是使用放射性同位素示踪剂和PBD6。在世界某些地区,包括发展中国家,这些示踪剂可能难以采购,寻找替代示踪剂对于解决这一问题至关重要。
最初,MBD被 Wong 等人用作映射哨兵节点 (SN)7的跟踪代理。在使用猫科模型的研究中,MBD的皮内注射显示淋巴摄取不良,而异硫丹蓝被选为哨兵节点(SN)映射7的首选染料。自从西蒙斯等人8日首次成功报告以来,甲基蓝染料已用于乳腺癌SNB。一些研究也报告MBD是SN鉴定的有利染料,MBD技术的假阴性率可与放射性同位素或PBD9、10、11相媲美。减少过敏反应和更低的价格是考虑在SN映射12中使用的其他原因。
最近,我们研究了在临床节点阴性乳腺癌中单独使用1%MBD对SNB。在早期阶段,MBD具有有利的识别率和负预测值13。如果我们在乳房的上外象限或奶嘴复合物 (NAC) 处出现外检活检疤痕,我们向下腹空间或腹肌区域注射 2 mL 的 1% MBD。蓝色节点和带淋巴蓝色通道的非蓝色节点被归类为 SN。在斧片的解剖地标被用作寻找SN的指导。 术中检查用于评估转移和SN被发送到组织病理学分析基于美国临床肿瘤学会(ASCO)指南14。
如果仔细选择病例,并且由外科医生和病理学家获得此技术所需的技能,许多患者可以挽救从ALND的有害影响,同时仍然有良好的生存。

<table border="0" cellpadding="0" cellspacing="0" style="width:1046px;" width="1046">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:348px;">Metiblo 50mg/5ml</td>
			<td style="width:174px;">Laboratories Sterop NV</td>
			<td style="width:353px;">BE475217</td>
			<td style="width:171px;">Methylene Blue Dye</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Disposable syringe with needle - 3mL (Luer Lock Tip)</td>
			<td>Terumo Europe NV</td>
			<td>dvr-3414</td>
			<td>Syringe for Injection</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">ForceTriad energy platform</td>
			<td>Medtronic</td>
			<td>ForceTriad</td>
			<td>Surgical cautery</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Shandon Cryomatrix embedding resin</td>
			<td>Thermo (scientific)</td>
			<td>6769006</td>
			<td rowspan="2">Frozen Section</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Cryotome FSE</td>
			<td>Thermo (scientific)</td>
			<td>77210153</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">HistoStar Embedding Workstation</td>
			<td>Thermo (scientific)</td>
			<td>A81000001</td>
			<td rowspan="3" style="width:171px;">Histopathological Examination</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Finesse Me+</td>
			<td>Thermo (scientific)</td>
			<td>A77510272</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Gemini AS</td>
			<td>Thermo (scientific)</td>
			<td>A81500002</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Benchmark GX</td>
			<td>Ventana Medical Systems</td>
			<td>750-850</td>
			<td rowspan="2">Immunohistochemistry</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Benchmark XT</td>
			<td>Ventana Medical Systems</td>
			<td>750-700</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Microscope</td>
			<td>Olympus</td>
			<td>Model BX53</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Ultrasound</td>
			<td>Phillips</td>
			<td>HD 7XE</td>
			<td></td>
		</tr>
	</tbody>
</table>

the application of 1% methylene blue dye as a single technique in breast cancer sentinel node biopsy

在这项研究中,我们向乳房的亚耳原或胸周空间注射了1%的亚甲基蓝色染料(MBD)。在吸乳手术(BCS)中,在下部斧头发线中做了一个单独的切口来查找哨点节点(SNs)。在乳房切除术中,SN通过同一乳房切除切口被识别。SN 被描述为蓝色节点或具有淋巴蓝色通道的节点。在斧片的解剖地标被用来促进SN识别。SNs转移是一个黄金标准,通过术中冷冻切片分析和组织病理学检查来评估。在这里,我们描述MBD是乳腺癌哨点节点活检(SNB)中唯一的技术,当无法提供放射性同位素示踪剂或专利或异硫酸盐蓝色染料(PBD)时,该技术可能很有用。

在这里,我们描述从呈现的技术的结果。两毫升1%MBD被注射在深亚罗拉空间作为注射的标准技术(图1A)。如果指示了微体注射,则应在超声波指导下进行(图1B)。进入突经空间后,可以看到蓝色结节或淋巴蓝色区域。继淋巴蓝片后,找到SN(图2A)。如果看不到蓝色结节或淋巴蓝片,我们使用闭周神经和侧胸静脉作为解?...

Watch this Scientific Journal Video about The Application of 1% Methylene Blue Dye As a Single Technique in Breast Cancer Sentinel Node Biopsy at JoVE.com

The Application of 1% Methylene Blue Dye As a Single Technique in Breast Cancer Sentinel Node Biopsy

在印度尼西亚,由于提供放射性同位素示踪剂和异硫酸盐或专利蓝色染料(PBD)的限制,乳腺癌手术不定期进行哨点节点活检(SNB)。为了克服这一障碍,我们应用1%亚甲基蓝色染料(MBD)作为单一剂来映射哨兵节点(SNs)。

1%亚甲蓝染料作为单一技术在乳腺癌哨点节点活检中的应用

In this study, we injected 1% methylene blue dye (MBD) into the subareolar or peritumoral space of the breast. In the case of breast conserving surgery ...

将百分之一甲基蓝染料作为单一技术应用于乳腺癌哨点活检。该程序的总体目标是将蓝色染料单独应用为早期乳腺癌轴结活检的单一技术，使用1%甲基蓝染料，并展示如何对哨点节点进行异硫素检测。第一节制备百分之一的甲苯蓝色染料及注射技术。第二节哨点节点活检技术在乳房保护手术中。第三节。乳腺切除术中的哨点点活检技术。第四节哨点节点病理检查。第五节术内检查。第六节结果。第七节结论。

the-application-1-methylene-blue-dye-as-single-technique-breast

Research

JoVE Journal

Cancer Research

18.3K 次观看.  Dharmais Cancer Hospital. 将百分之一甲基蓝染料作为单一技术应用于乳腺癌哨点活检。该程序的总体目标是将蓝色染料单独应用为早期乳腺癌轴结活检的单一技术，使用1%甲基蓝染料，并展示如何对哨点节点进行异硫素检测。第一节制备百分之一的甲苯蓝色染料及注射技术。第二节哨点节点活检技术在乳房保护手术中。第三节。乳腺切除术中的哨点点活检技术。第四节哨点节点病理检查。?...

视频: The Application of 1% Methylene Blue Dye As a Single Technique in Breast Cancer Sentinel Node Biopsy

Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures

The identification of functional driver events in cancer is central to furthering our understanding of cancer biology and indispensable for the discovery of the next generation of novel drug targets. It is becoming apparent that more complex models of cancer are required to fully appreciate the contributing factors that drive tumorigenesis in vivo and increase the efficacy of novel therapies that make the transition from pre-clinical models to clinical trials.
Here we present a methodology for generating uniform and reproducible tumor spheroids that can be subjected to siRNA functional screening. These spheroids display many characteristics that are found in solid tumors that are not present in traditional two-dimension culture. We show that several commonly used breast cancer cell lines are amenable to this protocol. Furthermore, we provide proof-of-principle data utilizing the breast cancer cell line BT474, confirming their dependency on amplification of the epidermal growth factor receptor HER2 and mutation of phosphatidylinositol-4,5-biphosphate 3-kinase (PIK3CA) when grown as tumor spheroids. Finally, we are able to further investigate and confirm the spatial impact of these dependencies using immunohistochemistry.

Identifying novel drug targets that transition from pre-clinical testing to human trials is a scientific priority. To that end, here we describe a functional genomics approach for examining the impact of gene depletion on cancer cell line spheroids, which more appropriately model human cancers in vivo.

利用功能基因组筛选识别在肿瘤细胞球体培养潜在的新药物靶标

The identification of functional driver events in cancer is central to furthering our understanding of cancer biology and indispensable for the discovery ...

科研

癌症研究

Ultrasonographic Evaluation of Breast Cancer-related Lymphedema

Lymphedema is one of the most common complications after breast cancer surgery. There are many diagnostic tools for lymphedema, but no standard method yet exists. Progressive Resistance Exercise (PRE) is expected to improve lymphedema without additional swelling. This study showed the therapeutic effects of PRE on lymphedema by using ultrasonography to measure the change in thickness of the muscle and subcutaneous tissue. The thickness of subcutaneous tissue decreased more in the PRE group than in the non-PRE group. Ultrasonography is widely used in many clinics because of its easy accessibility, safety, and inexpensiveness. Ultrasound is one of the best tools for diagnosing and determining treatment efficacy on breast cancer-related lymphedema (BCRL).

This study utilized ultrasonography for diagnosis and follow-up tests to confirm treatment efficacy of Progressive Resistance Exercise (PRE) in breast cancer-related lymphedema (BCRL). Ultrasonography can be applied effectively to ensure diagnosis and treatment of lymphedema.

乳腺癌相关的淋巴水肿的超声评价

Lymphedema is one of the most common complications after breast cancer surgery. There are many diagnostic tools for lymphedema, but no standard method yet ...

Studying Triple Negative Breast Cancer Using Orthotopic Breast Cancer Model

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less aggressive breast tumors, the 5-year survival rate of TNBC patients is 77% due to their characteristic drug-resistant phenotype and metastatic burden. Toward this end, murine models have been established aimed at identifying novel therapeutic strategies limiting TNBC tumor growth and metastatic spread. This work describes a practical guide for the TNBC orthotopic model where MDA-MB-231 breast cancer cells suspended in a basement membrane matrix are implanted in the fourth mammary fat pad, which closely mimics the cancer cell behavior in humans. Measurement of tumors by caliper, lung metastasis assessment via in vivo and ex vivo imaging, and molecular detection are discussed. This model provides an excellent platform to study therapeutic efficacy and is especially suitable for the study of the interaction between the primary tumor and distal metastatic sites.

This work presets an advanced protocol to accurately assess tumor loading by detection of green fluorescent protein and bioluminescence signals as well as the integration of quantitative molecular detection technique.

研究使用正交乳腺癌模型的三阴性乳腺癌

Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. When compared to patients with less ...

Orthotopic Transplantation of Breast Tumors as Preclinical Models for Breast Cancer

Preclinical models that faithfully recapitulate tumor heterogeneity and therapeutic response are critical for translational breast cancer research. Immortalized cell lines are easy to grow and genetically modify to study molecular mechanisms, yet the selective pressure from cell culture often leads to genetic and epigenetic alterations over time. Patient-derived xenograft (PDX) models faithfully recapitulate the heterogeneity and drug response of human breast tumors. PDX models exhibit a relatively short latency after orthotopic transplantation that facilitates the investigation of breast tumor biology and drug response. The transplantable genetically engineered mouse models allow the study of breast tumor immunity. The current protocol describes the method to orthotopically transplant breast tumor fragments into the mammary fat pad followed by drug treatments. These preclinical models provide valuable approaches to investigate breast tumor biology, drug response, biomarker discovery and mechanisms of drug resistance.

Patient-derived xenograft (PDX) models and transplantable genetically engineered mouse models faithfully recapitulate human disease and are preferred models for basic and translational breast cancer research. Here, a method is described to orthotopically transplant breast tumor fragments into the mammary fat pad to study tumor biology and evaluate drug response.

乳腺癌临床前模型乳腺肿瘤的矫形移植

Preclinical models that faithfully recapitulate tumor heterogeneity and therapeutic response are critical for translational breast cancer research. ...

Using Computer-based Image Analysis to Improve Quantification of Lung Metastasis in the 4T1 Breast Cancer Model

Breast cancer is a devastating malignancy, accounting for 40,000 female deaths and 30% of new female cancer diagnoses in the United States in 2019 alone. The leading cause of breast cancer related deaths is the metastatic burden. Therefore, preclinical models for breast cancer need to analyze metastatic burden to be clinically relevant. The 4T1 breast cancer model provides a spontaneously-metastasizing, quantifiable mouse model for stage IV human breast cancer. However, most 4T1 protocols quantify the metastatic burden by manually counting stained colonies on tissue culture plates. While this is sufficient for tissues with lower metastatic burden, human error in manual counting causes inconsistent and variable results when plates are confluent and difficult to count. This method offers a computer-based solution to human counting error. Here, we evaluate the protocol using the lung, a highly metastatic tissue in the 4T1 model. Images of methylene blue-stained plates are acquired and uploaded for analysis in Fiji-ImageJ. Fiji-ImageJ then determines the percentage of the selected area of the image that is blue, representing the percentage of the plate with metastatic burden. This computer-based approach offers more consistent and expeditious results than manual counting or histopathological evaluation for highly metastatic tissues. The consistency of Fiji-ImageJ results depends on the quality of the image. Slight variations in results between images can occur, thus it is recommended that multiple images are taken and results averaged. Despite its minimal limitations, this method is an improvement to quantifying metastatic burden in the lung by offering consistent and rapid results.

We describe a more consistent and expeditious method to quantify lung metastasis in the 4T1 breast cancer model by using Fiji-ImageJ.

利用计算机图像分析提高4T1乳腺癌模型中肺转移的定量

Breast cancer is a devastating malignancy, accounting for 40,000 female deaths and 30% of new female cancer diagnoses in the United States in 2019 alone. ...

Modeling Breast Cancer in Human Breast Tissue using a Microphysiological System

Breast cancer (BC) remains a leading cause of death for women. Despite more than $700 million invested in BC research annually, 97% of candidate BC drugs fail clinical trials. Therefore, new models are needed to improve our understanding of the disease. The NIH Microphysiological Systems (MPS) program was developed to improve the clinical translation of basic science discoveries and promising new therapeutic strategies. Here we present a method for generating MPS for breast cancers (BC-MPS). This model adapts a previously described approach of culturing primary human white adipose tissue (WAT) by sandwiching WAT between adipose-derived stem cell sheets (ASC)s. Novel aspects of our BC-MPS include seeding BC cells into non-diseased human breast tissue (HBT) containing native extracellular matrix, mature adipocytes, resident fibroblasts, and immune cells; and sandwiching the BC-HBT admixture between HBT-derived ASC sheets. The resulting BC-MPS is stable in culture ex vivo for at least 14 days. This model system contains multiple elements of the microenvironment that influence BC including adipocytes, stromal cells, immune cells, and the extracellular matrix. Thus BC-MPS can be used to study the interactions between BC and its microenvironment. 
We demonstrate the advantages of our BC-MPS by studying two BC behaviors known to influence cancer progression and metastasis: 1) BC motility and 2) BC-HBT metabolic crosstalk. While BC motility has previously been demonstrated using intravital imaging, BC-MPS allows for high-resolution time-lapse imaging using fluorescence microscopy over several days. Furthermore, while metabolic crosstalk was previously demonstrated using BC cells and murine pre-adipocytes differentiated into immature adipocytes, our BC-MPS model is the first system to demonstrate this crosstalk between primary human mammary adipocytes and BC cells in vitro.

This protocol describes the construction of an in vitro microphysiological system for studying breast cancer using primary human breast tissue with off the shelf materials.

使用微生理系统模拟人体乳房组织中的乳腺癌

Breast cancer (BC) remains a leading cause of death for women. Despite more than $700 million invested in BC research annually, 97% of candidate BC drugs ...

Modeling Brain Metastasis Via Tail-Vein Injection of Inflammatory Breast Cancer Cells

Metastatic spread to the brain is a common and devastating manifestation of many types of cancer. In the United States alone, about 200,000 patients are diagnosed with brain metastases each year. Significant progress has been made in improving survival outcomes for patients with primary breast cancer and systemic malignancies; however, the dismal prognosis for patients with clinical brain metastases highlights the urgent need to develop novel therapeutic agents and strategies against this deadly disease. The lack of suitable experimental models has been one of the major hurdles impeding advancement of our understanding of brain metastasis biology and treatment. Herein, we describe a xenograft mouse model of brain metastasis generated via tail-vein injection of an endogenously HER2-amplified cell line derived from inflammatory breast cancer (IBC), a rare and aggressive form of breast cancer. Cells were labeled with firefly luciferase and green fluorescence protein to monitor brain metastasis, and quantified metastatic burden by bioluminescence imaging, fluorescent stereomicroscopy, and histologic evaluation. Mice robustly and consistently develop brain metastases, allowing investigation of key mediators in the metastatic process and the development of preclinical testing of new treatment strategies.

We describe a xenograft mouse model of breast cancer brain metastasis generated via tail-vein injection of an endogenously HER2-amplified inflammatory breast cancer cell line.

通过炎症性乳腺癌细胞的尾静脉注射模拟脑转移

Metastatic spread to the brain is a common and devastating manifestation of many types of cancer. In the United States alone, about 200,000 patients are ...

Isolation of Primary Cancer-Associated Fibroblasts from a Syngeneic Murine Model of Breast Cancer for the Study of Targeted Nanoparticles

Cancer-associated fibroblasts (CAFs) are key actors in the context of the tumor microenvironment. Despite being reduced in number as compared to tumor cells, CAFs regulate tumor progression and provide protection from antitumor immunity. Emerging anticancer strategies aim to remodel the tumor microenvironment through the ablation of pro-tumorigenic CAFs or reprogramming of CAFs functions and their activation status. A promising approach is the development of nanosized delivery agents able to target CAFs, thus allowing the specific delivery of drugs and active molecules. In this context, a cellular model of CAFs may provide a useful tool for in vitro screening and preliminary investigation of such nanoformulations.
This study describes the isolation and culture of primary CAFs from the syngeneic 4T1 murine model of triple-negative breast cancer. Magnetic beads were used in a 2-step separation process to extract CAFs from dissociated tumors. Immunophenotyping control was performed using flow cytometry after each passage to verify the process yield. Isolated CAFs can be employed to study the targeting capability of different nanoformulations designed to tackle the tumor microenvironment. Fluorescently labeled H-ferritin nanocages were used as candidate nanoparticles to set up the method. Nanoparticles, either bare or conjugated with a targeting ligand, were analyzed for their binding to CAFs. The results suggest that ex vivo extraction of breast CAFs may be a useful system to test and validate nanoparticles for the specific targeting of tumorigenic CAFs.

This paper aims to provide a protocol for the isolation and culture of primary cancer-associated fibroblasts from a syngeneic murine model of triple-negative breast cancer and their application for the preclinical study of novel nanoparticles designed to target the tumor microenvironment.&#160;

将原发性癌症相关纤维细胞与乳腺癌的合成素 Murine 模型分离，用于靶向纳米粒子的研究

Cancer-associated fibroblasts (CAFs) are key actors in the context of the tumor microenvironment. Despite being reduced in number as compared to tumor ...

Mammary Epithelial and Endothelial Cell Spheroids as a Potential Functional In vitro Model for Breast Cancer Research

Breast cancer is the leading cause of mortality in women. The growth of breast cancer cells and their subsequent metastasis is a key factor for its progression. Although the mechanisms involved in promoting breast cancer growth have been intensively studied using monocultures of breast cancer cells such as MCF-7 cells, the contribution of other cell types, such as vascular and lymphatic endothelial cells that are intimately involved in tumor growth, has not been investigated in depth. Cell-cell interaction plays a key role in tumor growth and progression. Neoangiogenesis, or the development of vessels, is essential for tumor growth, whereas the lymphatic system serves as a portal for cancer cell migration and subsequent metastasis. Recent studies provide evidence that vascular and lymphatic endothelial cells can significantly influence cancer cell growth. These observations imply a need for developing in vitro models that would more realistically reflect breast cancer growth processes in vivo. Moreover, restrictions in animal research require the development of ex vivo models to elucidate better the mechanisms involved.
This article describes the development of breast cancer spheroids composed of both breast cancer cells (estrogen receptor-positive MCF-7 cells) and vascular and/or lymphatic endothelial cells. The protocol describes a detailed step-by-step approach in creating dual-cell spheroids using two different approaches, hanging drop (gold standard and cheap) and 96-well U-bottom plates (expensive). In-depth instructions are provided for how to delicately pick up the formed spheroids to monitor growth by microscopic sizing and assessing viability using dead and live cell staining. Moreover, procedures to fix the spheroids for sectioning and staining with growth-specific antibodies to differentiate growth patterns in spheroids are delineated. Additionally, details for preparing spheroids with transfected cells and methods to extract RNA for molecular analysis are provided. In conclusion, this article provides in-depth instructions for preparing multi-cell spheroids for breast cancer research.

Mammary Epithelial and Endothelial Cell Spheroids as a Potential Functional In vitro Model for Breast Cancer Research

Crosstalk between mammary epithelial cells and endothelial cells importantly contributes to breast cancer progression, tumor growth, and metastasis. In this study, spheroids have been made from breast cancer cells together with vascular and/or lymphatic endothelial cells and demonstrate their applicability as an in vitro system for breast cancer research.

乳腺上皮和内皮细胞球状体作为乳腺癌研究的潜在功能 体外 模型

Breast cancer is the leading cause of mortality in women. The growth of breast cancer cells and their subsequent metastasis is a key factor for its ...

Sentinel Lymph Node Mapping and Biopsy for Endometrial Cancer at Early Stage with Laparoscopy

Sentinel lymph node (SLN) mapping and biopsy is a promising technique for visualizing and evaluating lymph node status in cancer. This approach has been recommended for low-risk endometrial cancer (EC) patients by authoritative international guidelines, but it has not been performed broadly in China and worldwide. This work aims to describe detailed SLN mapping and biopsy procedures to promote the clinical application. SLN mapping and postoperative pathologic ultrastaging were conducted in a patient with low-risk EC using indocyanine green (ICG) dye to track the SLNs under laparoscopy and resecting them completely for ultrastaging. In conclusion, this protocol describes details of ICG injection, and SLN mapping and biopsy in EC patients based on the experiences gained during clinical practice.

This protocol describes the identification and resection of sentinel lymph nodes to make the operation as easy and minimally invasive as possible.

腹腔镜检查早期子宫内膜癌的前哨淋巴结定位和活检

Sentinel lymph node (SLN) mapping and biopsy is a promising technique for visualizing and evaluating lymph node status in cancer. This approach has been ...