저자는 공개 할 것이 없다.

인간 과목을 포함한 모든 절차는 040/KEPK/VII/2017의 인증서 번호로 달메이스 암 병원 윤리위원회의 승인을 받았습니다. 모든 환자는 동의서에 서명하고 이 연구에 참여하기로 동의를 표명했습니다.
참고 : 포함 기준은 종양 단계 T1-T2가 만져지지 않고 초음파 림프절 확대 (cNo)가없는 조기 유방암 진단을 가진 환자입니다. 배제 기준은 국소진행유방암, 신보조항화학요법 또는 호르몬요법, 및 임신이다.
1. 1 % 메틸렌 블루 염료 및 사출 기술 의 준비
<ol><li>환자를 마취 한 후 수술 장을 살균하십시오.</li>
	<li>3 mL 주사기와 함께 바이알에서 1 % 메틸렌 블루 염료의 2 mL을 흡인.</li>
	<li>가슴 주요 근육의 측면 테두리 아래 아래 의 작은 머리 라인을 표시하는 선을 그립니다.</li>
	<li>유방의 subareolar 공간에 1% 메틸렌 청색 염료의 2 mL를 주입하십시오.</li>
	<li>유방 또는 NAC의 상부 외부 사분면에 흉터가있는 경우, 선형 프로브 (12 MHz)를 가진 초음파 지도하에 23G 바늘로 흉터의 측면의 유방 자작 나무에 1 % 메틸렌 블루 염료의 2 mL을 주입하십시오.</li>
	<li>주사 부위에서 유방을 5 분 동안 원형으로 마사지 한 다음 수술을 계속하십시오.</li>
</ol>2. 유방 보존 수술에서 센티넬 노드 생검 기술 (BCS)
참고 : 수술은 BCS와 SNB를 받은 환자에서 수행됩니다.
<ol><li>단극 성 전기 캐스터리, DeBakey 집게 / 해부학 적 집게 및 리트랙터 : 수술 도구를 준비합니다.</li>
	<li>피부, 피하 조직 및 근막을 절개하십시오.</li>
	<li>파란색 노드 또는 파란색 림프관을 찾습니다. 림프파란색 관이 있는 파란색 노드 또는 파란색이 아닌 노드를 식별할 수 있을 때까지 파란색 관을 따라가십시오.</li>
	<li>파란색 노드 또는 파란색 림프관을 찾을 수없는 경우 늑간 신경 과 측면 흉부 정맥을 따라 센티넬 노드를 검색합니다.</li>
	<li>센티넬 노드를 조심스럽게 절제하고 노드가 손상되지 않도록 합니다.</li>
	<li>추가 의심 스러운 악성 림프절 확대를 찾기 위해 차 축 공간을 palpate.</li>
</ol>3. 유방 절제술의 센티넬 노드 생검 기술
참고 : 수술은 유방 절제술과 SNB를 받은 환자에서 이루어집니다.
<ol><li>피부와 피하 조직을 절개하십시오.</li>
	<li>스킨 플랩을 만듭니다.</li>
	<li>축사 fossa가 완전히 노출 될 때까지 가슴 주요에서 유방을 제거합니다.</li>
	<li>클라비코펙토랄 근막을 절개하여 센티넬 노드를 찾습니다.</li>
	<li>파란색 림프관을 찾을 수없는 경우, 늑간 신경과 측면 흉부 정맥 영역을 따라 파수 노드를 찾을 수 있습니다.</li>
	<li>센티넬 노드를 제거합니다.</li>
	<li>촉진에 의해 추가 의심 스러운 림프절에 대 한 보고.</li>
</ol>4. 수술 중 검사
<ol><li>긴 축에 평행하여 2mm보다 두껍지 않은 림프절을 슬라이스하십시오.</li>
	<li>각 노드에서 터치 각인 세포학을 합니다.</li>
	<li>외과용 시편을 금속 조직 디스크에 놓고 냉동 조직과 동일한 밀도의 젤형 배지에 포함시다.</li>
	<li>동결 된 섹션 (FS) 검사에 대 한 모든 노드를 제출 합니다.</li>
	<li>센티넬 노드의 전이성 상태를 양수 또는 음수로 분류하고 수술 중 외과 의사에게 보고합니다.</li>
</ol>5. 병리학적 검사
<ol><li>포르말린 고정 및 파라핀 내장 조직 절편에서 센티넬 노드의 최종 병리학 적 평가를 수행하십시오.</li>
	<li>암에 미국 합동위원회의 6 판에 따라 센티넬 노드 전이를 분류 (AJCC) 매뉴얼. 마크로전염(MAC)은 2 mm보다 큰 종양 침전물로 정의되며, 미세전이(MiC)는 0.2 및 2 mm 사이의 종양 침전물로 정의되며, 단리종양 세포(ITC)는 0.2 mm 보다 크지 않은 세포 클러스터로 정의된다.</li>
	<li>ITC 정의에 대한 의심이 있을 때 시토케라틴에 대한 직렬 단면 및 면역히스토화학 분석을 수행합니다.</li>
	<li>유사한 방법으로 차축 림프절의 나머지 부분을 검사합니다.</li>
</ol>

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우리는 무하마드, M.D. 준비하고 수술을 지원, 씨 알리 압둘 아지즈, 씨 아디티야 바유, 그리고 씨 아리아나 하리아디 원고 준비를 돕는. 우리는 또한 비디오 편집에 대한 그의 도움을 씨 테가 카리마를 인정합니다.

유방암 수술의 현대 시대에, SNB는 초기 유방암에 있는 차소 발판을 위한 배려의 표준으로 ALND를 대체하고 ALND는 SN이 전이에서 자유로운 경우에 포기되어야 합니다14,15. 선진국에서 일반적으로 사용되는 림프 매핑 기술은 방사성 동위원소 추적자 및 PBD를 결합 또는 단일 기술(16,17)으로적용한다. SNB를 수행하는...

차축 림프절 (ALN) 전이의 상태는 유방암에서 가장 중요한 예후 인자입니다. 차실 림프절 해부(ALND)는 ALNs1,2의전이성 상태를 평가하는 통상적인 절차였다. 불행히도, ALND는 이 절차 3,4후에 림프부종의 위험을 증가시킴으로써 삶의 질을 감소시키는 환자에게 이환율을 초래합니다. 요즘, 센티넬 노드 생검 (SNB)는 환자 중 최소한의 이환 때문에 차삭발판을 위한 ALND를 대체했습니다 5. SNB를 수행하는 가장 일반적인 방법은 방사성 동위원소 추적기및 PBD 6을 사용하는 것입니다. 개발도상국을 포함한 세계의 일부 지역에서는 이러한 트레이서 에이전트를 조달하기 어려울 수 있으며 문제를 해결하기 위해 대체 트레이서 에이전트를 검색하는 것이 중요합니다.
처음에 MBD는 Wong et al.에 의해 센티넬 노드(SNs)7을매핑하기 위한 트레이서 에이전트로 사용되었습니다. 고양이 모델을 이용한 그들의 연구에서, MBD의 상상 주입은 림프관 통풍관이 불량하고 이소술판 파랑이 센티넬 노드(SN)매핑에대한 바람직한 염료로서 선택되었다. 메틸렌 블루 염료는 시몬스 외. 8에 의해 첫번째 성공적인 보고부터유방암 SNB에서 이용되었습니다. 몇몇 연구 결과는 또한 SN 식별을 위한 유리한 염료로 MBD를 보고하고, MBD 기술의 거짓 부정비율은 방사성 동위 원소 또는 PBD 9,10,11에비교되었다는 것을. 적은 알레르기 반응과 낮은 가격은 SN 매핑(12)에서의사용을 고려해야하는 다른 이유입니다.
최근에, 우리는 임상적으로 노드 음성 유방암에 있는 SNB를 위한 1% MBD 단독의 사용을 공부했습니다. 초기 단계에서 MBD는 유리한 식별률과 음의 예측 값13을가합니다. 우리는 유방의 상부 외부 사분면 또는 젖꼭지 areolar 복합체 (NAC)에 절제생 흉터가 있는 경우에 subareolar 공간 또는 복막 지역에 1% MBD의 2 mL를 주사합니다. 림프 파란색 채널이 있는 파란색 노드와 파란색이 아닌 노드는 SN으로 분류됩니다. Axilla에 있는 해부학 적인 획기적인 것은 SN을 찾아기 위하여 지침으로 이용됩니다.
케이스가 신중하게 선택되고 이 기술에 요구되는 기술이 군의관에 의해 병리학자에 의해 장악되는 경우에, 많은 환자는 여전히 유리한 생존을 가진 동안 ALND의 유해한 효력에서 저장될 수 있었습니다.

<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

이 연구에서는, 우리는 주입 1% 메틸렌 블루 염료 (MBD) 유방의 subareolar 또는 복월적 공간에. 유방 보존 수술 (BCS)의 경우, 하부 액실라 헤어 라인에 별도의 절개가 센티넬 노드 (SNs)를 찾기 위해 만들어졌다. 유방 절제술에서, SN은 동일한 유방 절개를 통해 확인되었습니다. Ns는 림프청 채널이 있는 청색 노드 또는 노드로 설명되었습니다. 액실라의 해부학적 랜드마크는 SN 식별을 용이하게 하기 위해 사용되었습니다. SN 전이는 금 본위제이기 때문에 수술 중 동결 절편 분석 및 조직 병리학 검사에 의해 평가되었습니다. 여기서, 우리는 MBD를 방사성 동위 원소 추적자 또는 특허 또는 이소술판 청색 염료 (PBD)가 제공 될 수 없을 때 유용 할 수있는 유방암 센티넬 노드 생검 (SNB)에서 외로운 기술로 설명했다.

여기서는 제시된 기술의 결과를 설명합니다. 1% MBD의 2밀리리터를 주입의 표준 기법으로서 깊은 subareolar공간에 주입하였다(도 1A). 회액 주사가 표시되면 초음파 지도하에 수행해야합니다 (그림1B). 청색 노드 또는 림프 청색 관은 차축 공간을 입력 한 후 보였다. 림프청을 따라 면은 SN(그림2A)을찾는다. 파란...

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

인도네시아에서 센티넬 노드 생검(SNB)은 방사성 동위원소 트레이서 및 이소술판 또는 특허 청색 염료(PBD)를 제공하는 데 제한이 있기 때문에 유방암 수술을 위해 일상적으로 수행되지 않습니다. 이 장애물을 극복하기 위해 1% 메틸렌 블루 염료(MBD)를 단일 에이전트로 적용하여 센티넬 노드(SN)를 매핑했습니다.

유방암 센티넬 노드 생검에 있는 단 하나 기술로 1% 메틸렌 청색염의 응용

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 ...

유방암 센티넬 노드 생검의 단일 기술로 1% 메틸렌 블루 염료를 적용합니다. 이 절차의 전반적인 목표는 1%메틸렌 블루 염료를 사용하여 조기 유방암 축축노드 생검을 위한 단일 기술로서 단독으로 청염염의 적용을 보여주고 파수선 노드의 이소술판 검사를 하는 방법을 보여주는 것입니다. 1% 메틸렌 블루 염료및 사출 기술의 섹션 1 준비.섹션 2 센티넬 노드 생검 기술 유방 보존 수술. 섹션 3. 유방 절제술에서 센티넬 노드 생검 기술.파수선 노드의 4항 병리학 검사. 제5항 수술 중 검사. 섹션 6 결과.섹션 일곱 결론.

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

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JoVE Journal

Cancer Research

18.3K 조회수.  Dharmais Cancer Hospital. 유방암 센티넬 노드 생검의 단일 기술로 1% 메틸렌 블루 염료를 적용합니다. 이 절차의 전반적인 목표는 1%메틸렌 블루 염료를 사용하여 조기 유방암 축축노드 생검을 위한 단일 기술로서 단독으로 청염염의 적용을 보여주고 파수선 노드의 이소술판 검사를 하는 방법을 보여주는 것입니다. 1% 메틸렌 블루 염료및 사출 기술의 섹션 1 준비.섹션 2 센티넬 노드 생검 기술 유?...

비디오: 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.

기능 유전체학에게 검사를 활용하여 암 세포 회전 타원체 문화에 잠재적으로 소설 약 표적을 식별하는

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

연구

암 연구학

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.

유방암 관련 림프 부종의 초음파 평가

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.

정형외 유방암 모델을 사용하여 삼중 음성 유방암 을 연구

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.

유방암을 위한 전임상 모형으로 유방 종양의 정형소 이식

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.

4T1 유방암 모델에서 폐 전이의 정량화를 개선하기 위해 컴퓨터 기반 이미지 분석을 사용하여

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.

미생물 시스템을 사용하여 인간 유방 조직에서 유방암 모델링

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.

염증성 유방암 세포의 꼬리 정맥 주입을 통한 뇌 전이 모델링

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;

표적 나노 입자의 연구를 위한 유방암의 합성 Murine 모형에서 1 차적인 암 관련섬유세포의 격리

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.

유방암 연구를 위한 잠재적인 기능적인 시험관 내 모형으로 유방 상피 및 내피 세포 스페로이드

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.

복강경 검사와 초기 단계에서 자궁 내막암에 대한 센티넬 림프절 매핑 및 생검

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