Authors thank Jeffrey Sellers to assist orthotopic pancreatic cancer mouse modeling. This work was supported by Research Initiative Pilot Awards from the Department of Radiology at UAB and NIH grants 2P30CA013148 and P50CA101955.

모든 절차는 버밍엄 알라바마 대학의 기관 동물 관리 및 사용위원회에 의해 승인되었습니다. 1. 동소 이식 췌장 종양 마우스 모델링 <ol><li> 둘 베코 변성 이글 배지 (DMEM)에서 배양 표준 인간 췌장 암 세포주는 10 % 소 태아 혈청. 5 % CO 2 가습 분위기에서 37 ° C에서 모든 문화를 유지한다. </li><li> 8~10주 세 여자 심한 결합 면역 결핍 마우스를 사용합니다. 12 시간 빛과 12 시간의 어둠 사이클 RT에서 (21 ± 2 ° C의) 60 %의 습도에서 동물 케이지를 놓습니다. </li><li> 수술 내내 산소 (2 리터 / 분)과 혼합 된 이소 플루 란 2 %와 환기 장치를 사용하여 모든 동물을 마취. 발가락 핀치 반사에 의한 마취의 깊이를 확인합니다. 가열 패드 (37 ° C)에 도시 동물 체온을 유지한다. 마취 동안 건조를 방지하기 위해 눈에 수의학 안과 연고를 적용합니다. &amp;# 160; </li><li> 각 마우스의 복부의 왼쪽 윗부분에 머리를 제거하고 진통제를주는 영역으로 (5 ㎎ / 체중 피하의 kg carpofen). 노출 된 피부에 베타 딘 솔루션을 적용합니다. 멸균 수술 도구를 준비합니다. </li><li> 피부에 1cm의 절개를하고 홍채 바로 가위를 사용하여 복막. 조심스럽게 수술 핀셋을 사용하여 복부에서 췌장을 제거합니다. </li><li> 췌장의 꼬리에 0.5 ml의 인슐린 주사기의 28 G 바늘을 삽입하고 천천히 DMEM 30 μL에 250 만 인간 췌장암 세포의 용액을 주입. 작은 물집이 솔루션에 의해 췌장의 머리에서 생성되어 있는지 확인합니다. </li><li> 조심스럽게 다시 수술 핀셋을 사용하여 복부에 췌장을 배치합니다. (2) 5-0 prolene의 봉합을 중단과 1 층에 복막과 피부를 닫고 마취을 종료합니다. 완전히 회복 될 때까지 다른 동물의 회사에 수술을 한 동물을 반환하지 않습니다.60 7 ~ 10 일 후 수술에서 봉합사를 제거합니다. </li><li> 수술 후 24 시간에서 (carpofen, 5 ㎎ / 체중 피하의 kg) 진통제 약물의 또 다른 복용량을 제공합니다. </li><li> 두 손가락을 사용하여 수술 영역을 palpating하여 종양의 크기를 확인합니다. 종양은 일반적으로 주변 조직과 장기에 비해 밀도가 높고 울퉁불퉁 느낌. 종양을 느끼고 시작 2 주 - 보통 1 ~합니다. </li><li> 질병의 흔적을 매일 동물을 모니터링합니다. 동물 병 (부족 정상 정리 및 회피 행동)을 표시 할 때, 우리는 마취 상태에서 자궁 전위를 사용하여 종료되었습니다. </li></ol> 2. 자기 공명 영상 <ol><li> 세포 주입 후 보통 2 ~ 사주에 직경 7mm - 종양의 크기가 약 5을 때 MRI를 적용합니다. 작은 동물 이미징 또는 작은 동물 이미징을위한 전문 코일이 장착 된 임상 MR 스캐너 전용 MR 스캐너를 사용합니다. 참고 : H는 1 부피 공진기의 조합으로 9.4T 작은 동물 MR 스캐너를 사용/ 송신기 및 표면 코일 수신기 (직경 30mm) (브루 커 (Bruker) 사 BIOSPIN, 빌레 리카, MA). 표면 코일 나은 신호 대 잡음비 (SNR)를 제공한다 (14). </li><li> 0.2ml의 PBS (인산 완충 생리 식염수) - ~ 0.1 각각의 동물에게 0.2 밀리몰 / kg - ~ 0.1를 주입 가돌리늄 계 MRI 조영제를 준비한다. 참고 : 우리는 gadoteridol를 사용하고, 15 초 동안 (0.1 ㎖ / 초)를 통해 0.15 ml의 PBS 0.2 밀리몰 / kg을 주입. </li><li> 마이크로 polyethelene 튜브 (: 7.62 mm, 내경 : 0.28 mm, 외경 : 0.64 mm 길이)를 준비한다. 하나의 관의 단부와 다른 단부에 무딘 팁 30 G 바늘 (9.5 mm 길이)로 30 G 바늘 (12.7 mm 길이)를 삽입한다. 뭉툭한 팁 바늘에 MR 조영제를 포함하는 1 ML의 주사기를 연결하고 천천히 MR 조영제와 전체 튜브를 채우기 위해 주사기를 밀어 넣습니다. </li><li> 제제 전체에 산소 및 이미징 (2 L / 분)와 혼합 된 이소 플루 란 2 % - 1 ~ 환기를 사용 동물 마취. ANEST의 깊이를 확인발가락 핀치 반사에 의해 hesia. 마취 동안 건조를 방지하기 위해 눈에 수의학 안과 연고를 적용합니다. 니들 삽입 전에 가열 램프를 이용하여 지정 맥 꼬리를 팽창. 켈리 집게를 사용하여 30 G 바늘의 중간을 잡고, 꼬리 정맥에 삽입합니다. 플라스틱 또는 판지 종이 (10mm 폭 X 길이 100 mm)의 조각 위에 테이프를 모두 꼬리와 튜브 꼬리를 똑바로 유지합니다. </li><li> 촬영하는 동안 체온을 조절하기 위해 따뜻한 물 (또는 따뜻한 공기)를 순환 장착 동물 침대에 누운 위치에있는 동물을 놓습니다. 37 ° C에 침대에 온도를 설정합니다. 촬상 동안 체온을 모니터링하는 직장​​ 온도 프로브를 삽입한다. </li><li> 복부에 직각으로 구부러진 플라스틱 기판을 적용합니다. 종양이 보드의 상단 뒤에 위치되어 있는지 확인하고 약간 보드를 풀다운 (~ 2mm) 종양을 보장하기는 이사회가 발견된다. 단단히 동물 침대에 보드를 테이프입니다. </li><li> 호흡 패드 형질 도입을 테이프로가슴 부분에 R (SA 악기, 주식, 스토니 브룩, 뉴욕) 이미징 동안 동물의 호흡을 모니터링 할 수 있습니다. 종양 영역의 상단에 표면 코일을 놓고 단단히 동물 침대에 테이프. (: 내경 72mm) 볼륨 코일의 중심에 종양 영역을 배치 MR 스캐너로 동물 베드 누른다. </li><li> 수신기와 송신기 모두에 대해 매칭과 튜닝을 수행 shimming 하였다. </li><li> 종양을 찾을 수 해부학 MR 순서로 시작합니다. 다음 인수 매개 변수를 축 이미지를 얻기 위해 T2 강조 (T2W) 터보 스핀 에코 시퀀스를 사용합니다. 반복 시간 (TR) / 에코 시간 (TE) = 3000 / 34 밀리 초, 128 X 128 매트릭스,보기 30 X 30mm 필드, 평균 수 = 1, 에코 기차 길이 = 4, 20 연속 1mm 두께의 조각에 인터레이스 모드는 전체 종양 영역 (1.6 분 전체 주사 시간)을 커버한다. 주 : 소성 췌장 종양이 피하 것보다 위치되도록 더 어렵게하기 때문에, 종래 locali낮은 해상도의 ZER 이미지는 유용하지 않을 수 있습니다. </li><li> T1지도를 검색하기 위해 다양한 플립 각도 T1 강조 (T1W) 이미지를 획득. 평균 반복 시간 (TR) / 에코 시간 (TE) = 3분의 115 밀리 초, 128 X 128 매트릭스,보기의 30 X 30mm 필드 번호 :이를 위해, 다음 매개 변수를 multiflip 각도 접근 에코 그라데이션을 사용 = 4 ~ 5 - 종양 영역을 커버하는 인터레이스 모드에서 7 연속 1mm 두께의 슬라이스와 열 일곱 플립 각도, 20, 30, 40, 50, 60, 및 플립 각도 당 70 (총 스캔 시간 : 1 분). 주 : 그러나, multiflip 각도 접근법 B1 필드 균질성이 높은 경우에만 효과적이다. 그렇지 않으면, T1지도 여러 접근법 대신 TR (15)를 얻을 수있다. </li><li> 전과 MR 가돌리늄 계 조영제 주입 후에 T1W 이미지를 획득. ESP, K 공간의 중심이 획득되는 경우, 정상 상태를 보장하기 위해 T1 맵핑 (30)를 사용하지만, 선형 부호화의 고정 플립 각을 갖는 동일한 획득 파라미터 및 기하학을 사용하여짧은 TR과 낮은 수준의 플립 각도가 사용됩니다 ecially 때. 반면 주입 전에 5 기본 이미지를 획득. (45 분 전체 주사 시간)이어서 콘트라스트 주입 후 40의 이미지를 획득. 일정 속도 (0.01 ㎖ / 초)에 조영제를 주입 주사기 펌프를 사용한다. </li><li> 지속적으로 동물의 호흡을 모니터하고, 50 호흡을 유지하기 위해 이소 플루 란 농도를 조정 - 분당 100 호흡. 영상 전반에 걸쳐 동물의 체온을 모니터링합니다. </li><li> DCE-MRI를 완료 한 후, 바늘 및 기타 프로브를 벗고, 종이 수건과 침대 빈 새장에 동물을 배치합니다. 부드럽게 낮은 복부 마사지. 케이지는 복구로 동물 열 구배 내외부로 이동할 수 있도록 가열 램프 아래 절반에 배치되어야한다. 이 흉골 드러 누움을 유지하기 위해 충분한 의식을 회복 할 때까지 무인 동물을 방치하지 마십시오. </li></ol> 3. 이미지 처리 및 분석 <ol><li> 세그먼트 종양 정합<html> T2W 이미지에에. T2W 이미지에서, 종양 영역에서의 신호 강도는 주위 조직에 비해 더 밝은, 그래서 종양 경계를 수동으로 묘사 될 수있다. 주 : 이러한 글로벌 임계 또는 능동 윤곽 등 반자동 분할 기술 (16, 17)를 사용할 수 있지만, 배경 휘도 불균일은 코일면이 사용될 때 특히 보정 될 것이다. </li><li> T1과 양성자 밀도지도를 만듭니다. 그 에코 시간 (TE)를 가정 구배 에코 시퀀스 T1W 취득한 화상에서 T2 * 값은, 화소 값에 의해 결정되는 것보다 훨씬 적다 <img alt="식 (1)" fo:content-width="3in" src="/files/ftp_upload/52641/52641eq1.jpg" /> S 0 프로톤 밀도이고, T는 1 TR은 반복 시간이고, T 한 완화 시간 상수이며, θ는 플립 각도이다. 식 (1)에 다시 작성할 수 있습니다 641eq2.jpg &quot;/&gt; S (θ) / sinθ가 Y로 대체하고, S (θ) 때 / tanθ는 X로 대체됩니다. 수학 식 (2)는 선형 방정식이고, 그 기울기와 절편은 각각 T1 및 S 0 값을 검색하는 데 이용 될 수있다. </li><li> DCE-MR 영상에서 MR 대비 농도를 계산한다. MR 가돌리늄 계 조영제가 주입되면, T (1) 완화 시간 상수는 시간이 지남에 따라 변화된다. 따라서, 식 (1)에 다시 작성할 수 있습니다 <img alt="식 (3)" fo:content-width="3in" src="/files/ftp_upload/52641/52641eq3.jpg" /> 다음과 같이 T 1 (t)는, MR 대비 농도, C (t)와 관련이있다 <img alt="수학 식 4" fo:content-width="3in" src="/files/ftp_upload/52641/52641eq4.jpg" /> 여기서 R1은 MR 조영제의 길이 이완성이다. 그래서, 결합하여 식 (3) 및 (4), MR 조영제 농도에 의해 결정된다 <img aLT = &quot;식 5&quot;FO : 콘텐츠 폭 = &quot;3 인치&quot;SRC = &quot;/ 파일 / ftp_upload / 52641 / 52641eq5.jpg&quot;/&gt; </li><li> 콘트라스트 분사를 개시 한 후 시간 t에서 MR 콘트라스트 에이전트. C의 P (t)의 혈장 MR 조영제 농도를 나타낸다의 약동학 파라미터를 정량. C의 P (t하면) 동맥 입력 기능 (AIF)라고한다. AIF를 사용할 수있는 경우, MR 조영제의 약동학 적 파라미터에 의해 계산 될 수있다 <img alt="식 (6)" fo:content-width="4in" src="/files/ftp_upload/52641/52641eq6.jpg" /> C를 t (t)는 MR 조영제 농도가 표적 조직이고, V p를, V 전자 단편 혈관 외 세포 부피 분수 혈장 부피이고, K 트랜스 볼륨 전송 상수이다. 플럭스 속도 상수, k 개의 EP는 K 트란과 동일V 전자로 나눈 s의. AIF가없는 경우, 참조 영역 모델 대신에 사용될 수있다 (18,19). 기준 영역 모델은 흐름 제한 Kety 모델 (20)에 기초하여 다음과 같이된다 AIF 대한 필요성을 제거하는 기준 영역에 대비 농도를 사용하여, <img alt="식 (7)" fo:content-width="5in" src="/files/ftp_upload/52641/52641eq7.jpg" /> 여기서 C의 t, ROI (t), K 트랜스는, 투자 수익 (ROI) 및 V 전자는 ROI가, 관심 (ROI)의 영역에서 각각 대비 농도, 볼륨 전송 일정 및 분수 혈관 외 - 세포 외 볼륨입니다 C의 t 동안, RR (t), K 트랜스, RR, 및 V 즉, RR은 기준 영역의 것들이다. 척추 주위 근육은 종종 쥐 포장 된 V 전자, RR 참조 영역으로 선택하고,ravertebral 근육은 0.08 (21)에 일정하다고 가정한다. 우리는 기준 영역 모델을 사용 하였다. </li></ol>

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Authors do not have any conflicts of interest to declare.

우리는 면역 결핍 마우스, 마우스 복부 종양의 DCE-MRI, 그 운동 매개 변수의 정량화를 사용하여 소성 췌장 종양 모델링의 상세한 방법을 소개했다. 췌장의 꼬리에 바늘을 삽입 할 때 소성 췌장 종양 모델에서주의를 기울여야합니다. 성공적인 경우, 세포는 작은 소포를 만드는 췌장의 머리로 전달 될 것이다. 직각 굽힘 플라스틱 기판을 적용 할 때의 종양은 기판의 상부 단부 아래에 위치되는 것을 ?...

이 방법의 전반적인 목표는 생쥐에서 소성 췌장 종양 이종 이식에 대한 동적 대비 강화 된 자기 공명 영상 (DCE-MRI)을 적용하는 것입니다. DCE-MRI는 주사 후 일정 기간에 걸쳐 MR 콘트라스트의 변화를 모니터링함으로써 표적 조직에서 미세 혈관을 평가하는 비 침습적 방법이다. DCE-MRI는 악성 종양을 진단하고 치료 1-4 다양한 종양 반응을 평가하기 위해 이용되고있다. 양적 DCE-MRI는 높은 재현성 5를 발표했다. 표적 조직 MR 조영제의 약물 동태 학적 파라미터를 정량하기 위해, 콘트라스트 주입 전의 다양한 시점 및 T1 맵 취득한 모든 DCE-MR 이미지는 6 coregistered되어야한다. 그러나, 복부의 호흡과 연동 운동에 양적 DCE-MRI는 위장관 종양 제한된 응용 프로그램을했다. 동소 췌장 종양 모델을 평가하기 위해 사용되어왔다생물학적 치료와 화학 요법 7,8 다음 췌장 종양 반응. 원래 종양 부위에서의 미세 환경이 반사 및 치료함으로써 인간의 종양 반응을보다 정확하게 예측할 수 있기 때문에 종양 동소 이식 모델은, 종래의 피하 모델 우수 여겨진다. 그러나, 마우스의 췌장은 복부의 왼쪽 윗부분에 위치하고 있으며, 생쥐에서 소성 췌장 종양 이종 이식의 이렇게 양적 DCE-MRI는 쉽게 구현되지 않았습니다. 우리는 가슴 영역 (9)로부터 전송 동작을 방지하기 위해 직각 굽힘 플라스틱 기판을 사용하여 종양을 고정하여 마우스에서 복부 종양 DCE-MRI의 프로토콜을 확립했다. 이 보드에 적용되는 압력은 복부에 지역화하고, 호흡 곤란 결과 않았습니다. 자동 화상 coregistration 기술은 자유 호흡 모드 복부 장기의 DCE-MRI 대해 검증되었습니다 있지만 effectivel 수행Y는 단지 목표 영역은 규칙적으로 천천히 이동할 때 10. 동물의 호흡 속도는 이미징 동안 변수, 복부 너무 물리적 구속 정위 췌장 종양 마우스 모델에서 신뢰할 수있는 약물 동태 학적 매개 변수를 검색 할 필요가있다. 우리는 성공적 DCE-MRI 11-13 직각 굽힘 플라스틱 기판을 사용하여 소성 췌장 종양 이종 이식에서 MR 조영제의 약동학 파라미터를 정량 하였다. 여기에 우리가 동소 췌장 종양 모델링의 세부 절차를 제시, 종양 마우스에 이종 이식 및 약물 동태 학적 파라미터의 정량 DCE-MRI.

<table border="0" cellpadding="0" cellspacing="0" width="1275">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:245px;">Name of Material/ Equipment</td>
			<td style="width:199px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:712px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DMEM</td>
			<td>Invitrogen</td>
			<td>11965-118</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fetal bovine serum</td>
			<td>Harlan Laboratories</td>
			<td>BT-9501</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Betadine</td>
			<td style="width:199px;">Purdue products</td>
			<td style="width:119px;">67618-153-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">5-0 Prolene sutures</td>
			<td style="width:199px;">Ethicon</td>
			<td style="width:119px;">8720H</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">9.4T MR scanner</td>
			<td>Bruker Biospin Corporation</td>
			<td style="width:119px;">BioSpec 94/20 USR</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Gadoteridol</td>
			<td>Bracco Diagnostics Inc</td>
			<td style="width:119px;">NDC 0270-1111-03</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro-polyethelene tube</td>
			<td style="width:199px;">Strategic Applications, Inc</td>
			<td style="width:119px;">#PE-10-25</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">30G blunt tip needle</td>
			<td style="width:199px;">Strategic Applications, Inc</td>
			<td style="width:119px;">89134-194</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Monitoring and gating system</td>
			<td style="width:199px;">SA instruments, Inc</td>
			<td style="width:119px;">Model 1030</td>
			<td>This is an MR compatiable system to measure resiratory rating and body temperature of small animals at the same time.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Syringe pump</td>
			<td>New Era Pump Systems, Inc.</td>
			<td>NE-1600</td>
			<td></td>
		</tr>
	</tbody>
</table>

dynamic contrast enhanced magnetic resonance imaging of an orthotopic pancreatic cancer mouse model

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe respiratory motion artifact in the abdominal area. Orthotopic tumor models offer advantages over subcutaneous ones, because those can reflect the primary tumor microenvironment affecting blood supply, neovascularization, and tumor cell invasion. We have recently established a protocol of DCE-MRI of orthotopic pancreatic tumor xenografts in mouse models by securing tumors with an orthogonally bent plastic board to prevent motion transfer from the chest region during imaging. The pressure by this board was localized on the abdominal area, and has not resulted in respiratory difficulty of the animals. This article demonstrates the detailed procedure of orthotopic pancreatic tumor modeling using small animals and DCE-MRI of the tumor xenografts. Quantification method of pharmacokinetic parameters in DCE-MRI is also introduced. The procedure described in this article will assist investigators to apply DCE-MRI for orthotopic gastrointestinal cancer mouse models.

인간 췌장 종양 세포는 마우스 췌장 고형 종양 생성에 성공적으로 성장한다. (1) (A) 종양 세포 용액이 주입되어 정상적인 췌장, 및 (B)의 사진을 보여준다 동소 췌장 종양 이종 이식 베어링 대표적인 마우스 (MIA PACA -2- ). 종양은 다음 비장, 복부의 왼쪽 윗부분에 위치하고 있습니다. 세포 주입 후 직경 7 mm의 - 종양이 5 성장하는 사주 - 보통이 소요됩니다. <p class="jove_content...

Watch this Scientific Journal Video about Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model at JoVE.com

Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy.

동소 이식 췌장 암 마우스 모델의 동적 대비 강화 된 자기 공명 영상

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe ...

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Medicine

15.0K 조회수.  University of Alabama at Birmingham. The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy. 

비디오: Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

Contrast Enhanced Vessel Imaging using MicroCT

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between different soft tissues and body fluids is inherently poor in micro-CT images 1. Under these circumstances, visualization of blood vessels becomes a nearly impossible task. To overcome this and to improve the visualization of blood vessels exogenous contrast agents can be used. Herein, we present a methodology for visualizing the vascular network in a rodent model. By using a long-acting aqueous colloidal polydisperse iodinated blood-pool contrast agent, eXIA 160XL, we optimized image acquisition parameters and volume-rendering techniques for finding blood vessels in live animals. Our findings suggest that, to achieve a superior contrast between bone and soft tissue from vessel, multiple-frames (at least 5-8/ frames per view), and 360-720 views (for a full 360&deg; rotation) acquisitions were mandatory. We have also demonstrated the use of a two-dimensional transfer function (where voxel color and opacity was assigned in proportion to CT value and gradient magnitude), in visualizing the anatomy and highlighting the structure of interest, the blood vessel network. This promising work lays a foundation for the qualitative and quantitative assessment of anti-angiogenesis preclinical studies using transgenic or xenograft tumor-bearing mice.

Contrast enhanced small animal vessel imaging by microCT is a rapid, cost-effective and high-throughput technique for serial in situ examination for tumor development, for analyzing the network of blood vessels that nourish them, and for following the response of tumors to preclinical therapeutic intervention(s).

MicroCT를 사용하여 대비 향상된 베셀 영상

Microscopic computed tomography (microCT) offers high-resolution volumetric imaging of the anatomy of living small animals. However, the contrast between ...

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의학

An Orthotopic Model of Murine Bladder Cancer

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and subsequent cell adhesion. After their bladders are transurethrally catheterized and drained, animals are again catheterized to permit insertion of a platinum wire into bladders without damaging the urethra or bladder. The catheters are made of Teflon to serve as an insulator for the wire, which will conduct electrical current into the bladder to create a burn injury. An electrocautery unit is used to deliver 2.5W to the exposed end of the wire, burning away extracellular layers and providing attachment sites for carcinoma cells that are delivered in suspension to the bladder through a subsequent catheterization. Cells remain in the bladder for 90 minutes, after which the catheters are removed and the bladders allowed to drain naturally. The development of tumor is monitored via ultrasound. Specific attention is paid to the catheterization technique in the accompanying video.

Bladder tumors are established in female mice in a minimally invasive fashion through catheterization, local cauterization, and subsequent adhesion of carcinoma cells to the burn sites.

Murine 방광 암의 Orthotopic 모델

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

구강 편평 세포 암종의 Orthotopic 마우스 모델에서 종양 세포 침략의 다중 광자 이미징

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the understanding of the mechanics of tumor invasion, and direct intracerebral inoculation of tumor provides the opportunity of observing the invasive process in a physiologically appropriate environment2. As far as human brain tumors are concerned, the orthotopic models currently available are established either by stereotaxic injection of cell suspensions or implantation of a solid piece of tumor through a complicated craniotomy procedure3. In our technique we harvest cells from tissue culture to create a cell suspension used to implant directly into the brain. The duration of the surgery is approximately 30 minutes, and as the mouse needs to be in a constant surgical plane, an injectable anesthetic is used. The mouse is placed in a stereotaxic jig made by Stoetling (figure 1). After the surgical area is cleaned and prepared, an incision is made; and the bregma is located to determine the location of the craniotomy. The location of the craniotomy is 2 mm to the right and 1 mm rostral to the bregma. The depth is 3 mm from the surface of the skull, and cells are injected at a rate of 2 &mu;l every 2 minutes. The skin is sutured with 5-0 PDS, and the mouse is allowed to wake up on a heating pad. From our experience, depending on the cell line, treatment can take place from 7-10 days after surgery. Drug delivery is dependent on the drug composition. For radiation treatment the mice are anesthetized, and put into a custom made jig. Lead covers the mouse's body and exposes only the brain of the mouse. The study of tumorigenesis and the evaluation of new therapies for GBM require accurate and reproducible brain tumor animal models. Thus we use this orthotopic brain model to study the interaction of the microenvironment of the brain and the tumor, to test the effectiveness of different therapeutic agents with and without radiation.

The purpose of this article is to describe the use of an orthotopic glioblastoma model for chemoradiation studies. This article will go though cell processing, implanting, and radiotherapy of the mouse using an intracranial model.

Orthotopic 마우스 뇌종양 모델의 조합 방사선 치료

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the ...

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

자기 공명 영상, 체적 계산의 Tomography 및 초음파를 이용한 유방암의 뼈 전이의 누드 쥐 모델에서 Angiogenesis의 멀티 모달 이미징

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

An Orthotopic Mouse Model of Anaplastic Thyroid Carcinoma

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer cells into immunodeficient mice. Subcutaneous xenograft models have been valuable for preclinical screening and evaluation of new therapeutic treatments. There are a number of advantages to using a subcutaneous model; 1) rapid, 2) reproducible, and 3) tumor establishment, growth, and response to therapeutic agents may be monitored by visual inspection. However, substantial evidence has shed light on the short-comings of subcutaneous xenograft models1-3. For instance, medicinal treatments demonstrating curative properties in subcutaneous xenograft models often have no notable impact on the human disease. The microenvironment of the site of xenographic transplantation or injection lies at the heart of this dissimilarity.
Orthotopic tumor xenograft models provide a more biologically relevant context in which to study the disease. The advantages of implanting diseased cells or tissue into their anatomical origin equivalent within a host animal includes a suitable site for tumor-host interactions, development of disease-related metastases and the ability to examine site-specific influence on investigational therapeutic remedies. Therefore, orthotopic xenograft models harbor far more clinical value because they closely reproduce human disease. For these reasons, a number of groups have taken advantage of an orthotopic thyroid cancer model as a research tool4-7.
Here, we describe an approach that establishes an orthotopic model for the study of anaplastic thyroid carcinoma (ATC), which is highly invasive, resists treatment, and is virtually fatal in all diagnosed patients. Cultured ATC cells are prepared as a dissociated cellular suspension in a solution containing a basement membrane matrix. A small volume is slowly injected into the right thyroid gland. Overall appearance and health of the mice are monitored to ensure minimal post-operative complications and to gauge pathological penetrance of the cancer. Mice are sacrificed at 4 weeks, and tissue is collected for histological analysis. Animals may be taken at later time-points to examine more advance progression of the disease. Production of this orthotopic mouse model establishes a platform that accomplishes two objectives: 1) further our understanding of ATC pathology, and 2) screen current and future therapeutic agents for efficacy in combating ATC.

Generation of an orthotopic mouse model of anaplastic thyroid carcinoma is described here. This technique employs surgical placement of human anaplastic thyroid cancer cells into the thyroid of immunodeficient mice, thus creating a more clinically relevant setting to study disease progression as well as screen innovative therapeutic interventions.

역 형성 갑상선 암의 동소 마우스 모델

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer ...

An Orthotopic Bladder Cancer Model for Gene Delivery Studies

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are needed. The tumor microenvironment can significantly influence tumor development and therapy response. It is therefore often desirable to grow tumor cells in the organ from which they originated. This protocol describes an orthotopic model of bladder cancer, in which MB49 murine bladder carcinoma cells are instilled into the bladder via catheterization. Successful tumor cell implantation in this model requires disruption of the protective glycosaminoglycan layer, which can be accomplished by physical or chemical means. In our protocol the bladder is treated with trypsin prior to cell instillation. Catheterization of the bladder can also be used to deliver therapeutics once the tumors are established. This protocol describes the delivery of an adenoviral construct that expresses a luciferase reporter gene. While our protocol has been optimized for short-term studies and focuses on gene delivery, the methodology of mouse bladder catheterization has broad applications.

Implantation of cancer cells into the organ of origin can serve as a useful preclinical model to evaluate novel therapies. MB49 bladder carcinoma cells can be grown within the bladder following intravesical instillation. This protocol demonstrates catheterization of the mouse bladder for the purpose of tumor implantation and adenoviral delivery.

유전자 전달 연구 동소 방광암 모델

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are ...

Bioluminescent Orthotopic Model of Pancreatic Cancer Progression

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%. New therapeutic options are critically needed and depend on improved understanding of pancreatic cancer biology. To better understand the interaction of cancer cells with the pancreatic microenvironment, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression. Luciferase-tagged pancreatic cancer cells are resuspended in Matrigel and delivered into the pancreatic tail during laparotomy. Matrigel solidifies at body temperature to prevent leakage of cancer cells during injection. Primary tumor growth and metastasis to distant organs are monitored following injection of the luciferase substrate luciferin, using in vivo imaging of bioluminescence emission from the cancer cells. In vivo imaging also may be used to track primary tumor recurrence after resection. This orthotopic model is suited to both syngeneic and xenograft models and may be used in pre-clinical trials to investigate the impact of novel anti-cancer therapeutics on the growth of the primary pancreatic tumor and metastasis.

Improved understanding of pancreatic cancer biology is critically needed to enable the development of better therapeutic options to treat pancreatic cancer. To address this need, we demonstrate an orthotopic model of pancreatic cancer that permits non-invasive monitoring of cancer progression using in vivo bioluminescence imaging.

췌장암의 진행 발광 동소 모델

Pancreatic cancer has an extremely poor five-year survival rate of 4-6%.  New therapeutic options are critically needed and depend on improved ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

인간의 전립선 암 전이의 동소 쥐과 모델

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics disturbance in affected cells. Diffusion weighted magnetic resonance imaging (MRI) identifies regions that are damaged, potentially irreversibly, by hypoxia-ischemia. Alterations in glucose utilization in the affected tissue may be detectable by positron emission tomography (PET) imaging of 2-deoxy-2-(18F)fluoro-&#7429;-glucose ([18F]FDG) uptake. Due to the rapid and variable nature of injury in this animal model, acquisition of both modes of data must be performed simultaneously in order to meaningfully correlate PET and MRI data. In addition, inter-animal variability in the hypoxic-ischemic injury due to vascular differences limits the ability to analyze multi-modal data and observe changes to a group-wise approach if data is not acquired simultaneously in individual subjects. The method presented here allows one to acquire both diffusion-weighted MRI and [18F]FDG uptake data in the same animal before, during, and after the hypoxic challenge in order to interrogate immediate physiological changes.

The method presented here uses simultaneous positron emission tomography and magnetic resonance imaging. In the cerebral hypoxia-ischemia model, dynamic changes in diffusion and glucose metabolism occur during and after injury. The evolving and irreproducible damage in this model necessitates simultaneous acquisition if meaningful multi-modal imaging data are to be acquired.

마우스 대뇌 저산소증 - 허혈 동안 동시 PET / MRI 영상

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics ...