KF 및 KO는 KS와 함께 첫번째 저자 공동 아르 이 작품은 AHPBA ​​원정대 (KS), NIH K23 CA148964-01 (LZ), 존스 홉킨스 의과 대학 임상 과학 자상 (LZ), Viragh 재단과 존스 홉킨스 건너 뛰기 Viragh 췌장 암 센터 (EMJ에 의해 부분적으로 지원 LZ), 국립 췌장 재단 (LZ), Lefkofsky 가족 재단 (LZ), 위장관 암의 P50의 CA062924에서 NCI 포자 (EMJ, LZ), Lustgarten 재단 (LZ), 그리고 솔 골드만 췌장 암 센터의 보조금 (KS , BHE, LZ).

모든 동물 실험은 존스 홉킨스 대학의 동물 관리 및 사용위원회의 가이드 라인을 따른다, 동물은 평가 및 실험실 관리의 인증 (AAALAC)에 대한 협회의 가이드 라인에 따라 유지 하였다. 수술은 시간당 십오 공기 교류 최소 거친다 수술실에서 멸균 조건 하에서 수행된다. 모든 악기는 절차 중에 전에 각 마우스 사이에서 70 % 에탄올로 순서에 다시 오토 클레이브에 의해 살균된다. 수술 살아남을 것으로 예상되지 않은 모든 마우스는 마취 여전히 동안 자궁 경부 전위 다음 3 분 동안 CO 2에서 안락사된다. 1 셀 준비 <ol><li> 이 X10 7 세포 / ml에서 인산염 완충 식염수 Panc02 배양 세포를 재현 탁하고 얼음에 보관. </li></ol> 2 마우스 준비 <ol><li> Ketamin를 관리자일 라진 (xylazine) (10 ㎎ / ㎏)와 혼합 전자 (100 ㎎ / ㎏)을 복강에 8-12주 나누어 오래 된 C57BL / 6 마​​우스 여성. </li><li> 발가락 핀치 금단 반사가 마우스가 완전히 마취되어 있는지 확인하기 위해 폐지되어 있는지 확인하십시오. </li><li> 완전히 쥐를 마취 할 필요 자일 라진 (xylazine) (10 ㎎ / ㎏)와 혼합 케타민의 추가 용량 (100 ㎎ / ㎏)을 관리합니다. </li><li> 쥐가 마취 동안 건조를 방지하기 위해 쥐의 눈에 Puralube 수의사 연고를 적용합니다. </li><li> 상처의 오염을 방지하기 위해 마우스의 외과 영역을 면도. </li><li> 요오드 후 70 % 에탄올과 함께 절개의 좌측 영역 subcostal 수험. </li><li> 불임을 유지하기 위해 복부 수술 드레이프를 놓습니다. </li></ol> 3 개복술 <ol><li> 피부를 통해와 메스를 사용하여 복막을 통해 왼쪽 귀 라인에 왼쪽 subcostal 절개를합니다. </li><li> 동시에 디지털 페이지를 적용하여 절개를 통해 비장을 표현절개의 뇌와 꼬리 부분을 따라 ressure. </li><li> 비장의 하부 끝에 비장 혈관을 찾습니다. </li><li> 비장 극의 비장 혈관이 손상되지 않도록주의하면서 비장의 중심이 지평선 중간 크기 결찰 클립을 배치하여 비장을 나눈다. </li><li> 오염을 방지하기 위해 다시 복막에 비장의 상극을 놓습니다. </li></ol> 4 종양 주입 <ol><li> 26 G X 8 &quot;주사기에 인산염 완충 생리 식염수 150 μL를 그립니다. </li><li> 주입하는 동안 항상 수직 위치에 주사기를 유지 같은 주사기로 100 Panc02 ㎕의 (2 X10 6) 세포를 그립니다. </li><li> 70 % 에탄올 용액에서 바늘을 찍어. 바늘은 멸균을 위해 70 % 에탄올에 침지된다. </li><li> 주사기가 항상 수직으로 유지되어 있는지 확인되는 노출 hemispleen로 천천히 세포를 주입한다. syrin의 경사GE는하에 비장 세포를 주입 피하기 위해 비장 캡슐을 항상 지켜 져야한다. 비장과 혈관의 미백 주사에 관찰해야한다. </li><li> 비장을 상승 및 비장 혈관을 폐색하는 비장에서 한 매체 지평선 클립을 적용합니다. </li><li> 췌장 및 비장 혈관의 가장 후방 측면에 하나의 작은 지평선 클립을 적용합니다. </li><li> 지평선 클립 위에 절단하여 hemispleen에서 췌장 및 비장 혈관을 결찰. </li><li> 그 &#39;측에 마우스를 놓고 증류수로 절개를 관개. </li></ol> 마취에서 5 복부 폐쇄 및 복구 <ol><li> 4-0 실행 스티치로 복막을 닫습니다. </li><li> 피부 절개를 닫 2-3 피부 클립을 적용합니다. </li><li> 포스트 수술 통증을 완화하는 0.1 ㎎ / ㎏ 부 프레 노르 핀 또는 5 ~ 10 ㎎ / ㎏ Carprofen 피하을 관리 할 수​​ 있습니다. </li><li> REC위한 가열 패드에 마우스를 놓고overy 이동까지 마우스가 규칙적인 호흡 패턴을 보여줍니다. 동물은 완전히 수술에서 회복 한 후 다른 동물의 회사에 반환됩니다. </li></ol> 6 부검 검사 및 간의 수확 <ol><li> 수술 후 30-60일 사이에, 마우스는 전이의 임상 증상을 표시하기 시작하고 안락사가 필요합니다. 인도적인 안락사를 요구하는 전이성 질병의 징후 확대 복부와 복수의 개발을 포함한다. CO 2의 흡입에 의해 쥐를 안락사. </li><li> 흡입 후, 자궁 경부 전위를 수행합니다. </li><li> 동물이 아닌 응답 것으로 판정 된 후, 가위를 사용하여 복부를 노출 복막을 통해 절개. </li><li> 집게와 가위를 사용하여 조심스럽게 복부에서 간을 잘라. </li><li> OCT 화합물의 간을 놓고 플래시는 액체 질소를 사용하여 동결. -80 ° C에서 간을 저장합니다. 대안 적으로, 라이브R은 내장 24 시간 파라핀 실온에서 16 % 중성 포르말린에 고정 될 수있다. </li></ol>

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저자는 공개 관심은 아무런 충돌이 없다.

hemispleen 주입 기법을 통해 간으로 췌장 암전이 전임상 뮤린 모델간에 전이성 질환이있는 환자의 임상 적, 면역 학적 조건 중 많은 거울 설명한 제 모델이다. 이 모델은 다른 쥐 모델에 비해 여러 가지 장점을 제공합니다. 쥐의 30 %가 간 및 전이 형성의 타이밍에 전이를 개발하는 췌장암의 형질 전환 모델은 상당히 가변적 달리 먼저,이 모델에서 마우스의 100 %에서 간으로 일관 획득 전이 이점을 제?...

췌장 유관 선암 (PDA)는 미국 한 암 관련 사망의 네 번째 주요 원인이다. 전이성 PDA 환자 5 년 생존율은 2-12% 일이다. 췌장암에 대한 보조 및 보조 항암 화학 요법이 그 어느 때보 다 오늘날 더 효과적이지만, 여전히 새로운 치료를위한 절박한 필요성이 남아있다. 신규 한 치료제의 개발을 신뢰성 전임상 동물 모델을 필요로한다. 피하 종양 모델은 사용하기 간단하지만, 단점은 인간의 암에서 볼 수있는 종양의 진행과 미세 환경을 전이하고 모방하는이 종양의 무능력을 포함한다. 같은 Kras 호텔 및 p53의 노크에 발암 돌연변이 (KPC 모델) (2)와 YFP 계보 추적 (PKCY 모델)와 KPC 모델을 포함하는 유전자 변형 마우스와 같은 유전자 조작 모델, 3 immunocompete에서 자연적으로 발생하는 종양에 대한 연구 허용NT 마우스. 또한, 종양 미세 환경은 불변이고 더 밀접 인간 종양 진행에서 본 것을 닮았다. 불행하게도, 종양 발생의 타이밍 어려운 실험 요법을 연구 할 수있다 이러한 모델 내의 변수이다. 또한,이 마우스의 교잡 시간과 항상 가능하지 않을 금융 약속, 상당한 양을 필요로한다. 마지막으로, 이식 이종 이식 PDA 마우스 모델을 변경 한 마우스, 결석, 종양 미세 환경을 면역이 필요로. 따라서 이러한 전임상 모델에서 입증 실험 요법의 효능은 종종 인간 임상 시험에서 4 재현성 아니다. 이 hemisplenectomy 마우스 모델 C57BL / 6 마우스 (5, 6)로부터 유도 고도로 종양, 췌장 종양 methylcholanthrene 유도 세포주이다 Panc02 종양 세포를 이용한다. 또한, 다른 PDA 뮤린 세포주 KPC 마이크에서 파생전자는이 모델에서 사용될 수있다. Schulick 등. 이전에 hemisplenectomy 기술을 개발하고 대장 암의 전이 (7)의 동계 마우스 모델을 만들었습니다. 이 hemisplenectomy 모델 (그림 1)는 새로운 치료제 7-10의 조사 자체를 빌려주는 면역 생쥐에서 일관되고 동일한 종양 접종을 제공합니다.

<table border="0" cellpadding="0" cellspacing="0" width="713">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Cell culture media and components</td>
			<td style="width:197px;"></td>
			<td style="width:119px;"></td>
			<td style="width:181px;">will vary depending on cell line</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Phosphate Buffered Saline</td>
			<td style="width:197px;">Gibco by Life Technologies</td>
			<td style="width:119px;">10010-023</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">15 mL centrifigure tubes</td>
			<td style="width:197px;">Corning</td>
			<td style="width:119px;">430052</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Curved Operating Scissor</td>
			<td style="width:197px;">Roboz Surgical Store</td>
			<td style="width:119px;">RS-6835</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Micro dissecting Graefe Forceps</td>
			<td style="width:197px;">Roboz Surgical Store</td>
			<td style="width:119px;">RS-5130</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Needle holder (regular)</td>
			<td style="width:197px;">World Precision Instruments, Inc</td>
			<td style="width:119px;"></td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">3-0 or 4-0 suture</td>
			<td style="width:197px;"></td>
			<td style="width:119px;"></td>
			<td style="width:181px;">courtesy of dept of surgery, Johns Hopkins</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Weck Horizon Open Ligating Clip Applier, small, angled</td>
			<td style="width:197px;">Teleflex</td>
			<td style="width:119px;">137085</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Weck Horizon Open Ligating Clip Applier, medium, angled</td>
			<td style="width:197px;">Teleflex</td>
			<td style="width:119px;">237115</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Weck Horizon medium ligating clips</td>
			<td style="width:197px;">Teleflex</td>
			<td style="width:119px;">002200</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Weck Horizon small ligating clips</td>
			<td style="width:197px;">Teleflex</td>
			<td style="width:119px;">001200</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Syringe, 1 cc, tb syringe, 3/8&#34; slip tip</td>
			<td style="width:197px;">Becton Dickinson</td>
			<td style="width:119px;">309625</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Syringe, 1 cc, tb syringe, 5/8&#34; slip tip</td>
			<td style="width:197px;">Becton Dickinson</td>
			<td style="width:119px;">309597</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">9 mm Autoclip Applier</td>
			<td style="width:197px;">Mikron</td>
			<td style="width:119px;">427630</td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">9 mm Autoclip</td>
			<td style="width:197px;">Becton Dickinson</td>
			<td style="width:119px;">427631</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Heating pad</td>
			<td style="width:197px;">Sunbeam</td>
			<td style="width:119px;">CAT93</td>
			<td style="width:181px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">70% Ethanol</td>
			<td style="width:197px;"></td>
			<td style="width:119px;"></td>
			<td style="width:181px;">sold by numerous vendors</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ketamine Hydrochloride&#160;</td>
			<td style="width:197px;">Hospira</td>
			<td style="width:119px;">22395DD</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Xylazine hydrochloride</td>
			<td style="width:197px;">Sigma</td>
			<td style="width:119px;">X1251</td>
			<td></td>
		</tr>
	</tbody>
</table>

a preclinical murine model of hepatic metastases

수많은 뮤린 모델은 인간의 암 연구 및 암 치료 및 개발의 이해를 발전하기 위해 개발되어왔다. 여기서, 동계 생쥐 췌장 종양 세포의 주입 hemispleen 통한 간내 전이 전임상, 쥐 췌장 종양 모델을 설명한다. 이 모델간에 전이성 질환을 가진 환자에서의 임상 조건의 많은 모방. 마우스는 지속적으로 전이 과정의 조사, 실험적인 치료 테스트 및 종양 면역학 연구를 허용 간에서 전이을 개발한다.

hemispleen에 주입 Panc02 종양 세포 기술이 제대로 수행되는 경우 폐, 복막 전이가 관찰되지 않은 반면 (그림 1) 마우스의 100 %의 형태로 간 전이. 이 기술은 여러 개의 독립적 인 실험에서 Panc02 세포를 이용한 백 위에 마우스에 대해 수행되었으며, 재현성, 모든 미처리 마우스 hemisplenectomy 절차 (도 2) 다음 30 60 일 사이의 간전으로 죽는다. 치료 및 치료 그룹 사이에 유의 한 차이...

Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

A Preclinical Murine Model of Hepatic Metastases

간 전이 전임상, 뮤린 모델 hemispleen 주입 기법을 통해 수행 하였다.

간 전이의 전임상 쥐과 모델

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

a-preclinical-murine-model-of-hepatic-metastases

Research

JoVE Journal

Medicine

28.3K 조회수.  The Johns Hopkins University School of Medicine.  간 전이 전임상, 뮤린 모델 hemispleen 주입 기법을 통해 수행 하였다. 

비디오: A Preclinical Murine Model of Hepatic Metastases

Murine Bioluminescent Hepatic Tumour Model

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are administered specifically to the liver to induce a localised liver tumour, via mobilisation of the spleen and splitting into two, leaving intact the vascular pedicle for each half of the spleen. Lewis lung carcinoma cells that constitutively express the firefly luciferase gene (luc1) are inoculated into one hemi-spleen which is then resected 10 minutes later. The other hemi-spleen is left intact and returned to the abdomen. Liver tumour growth can be monitored by bioluminescence imaging using the IVIS whole body imaging system. Quantitative imaging of tumour growth using IVIS provides precise quantitation of viable tumour cells. Tumour cell death and necrosis due to drug treatment is indicated early by a reduction in the bioluminescent signal. This mouse model allows for investigating the mechanisms underlying metastatic tumour-cell survival and growth and can be used for the evaluation of therapeutics of liver metastasis. 

This article describes a procedure for the induction of orthotopic bioluminescent liver tumours in mice, and subsequent analysis of tumour growth confined to the liver using live whole body luminescence imaging.

Murine 발광 생물 간장 Tumour 모델

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are ...

연구

의학

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety.
Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential.
In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates.

A method of gene transfer for the treatment of ischemic heart failure is described using a swine model of myocardial infarction. Our simple and reproducible method enables us to readily evaluate the efficacy of various gene transfers with a very simple and reproducible way.

잠복기 모델에서 허혈성 심장 마비에 대한 유전자 전송

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

상처 치유의 생쥐 모델

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

지주막 하 출혈의 생쥐 모델

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

비 침습적 영상에 대한 생쥐 모델 검색 및 모니터 난소 암 재발

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Ex Vivo Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics

The molecular analysis of established cancer cell lines has been the mainstay of cancer research for the past several decades. Cell culture provides both direct and rapid analysis of therapeutic sensitivity and resistance. However, recent evidence suggests that therapeutic response is not exclusive to the inherent molecular composition of cancer cells but rather is greatly influenced by the tumor cell microenvironment, a feature that cannot be recapitulated by traditional culturing methods. Even implementation of tumor xenografts, though providing a wealth of information on drug delivery/efficacy, cannot capture the tumor cell/microenvironment crosstalk (i.e., soluble factors) that occurs within human tumors and greatly impacts tumor response. To this extent, we have developed an ex vivo (fresh tissue sectioning) technique which allows for the direct assessment of treatment response for preclinical and clinical therapeutics development. This technique maintains tissue integrity and cellular architecture within the tumor cell/microenvironment context throughout treatment response providing a more precise means to assess drug efficacy.

Ex Vivo Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics

Established cancer cell lines and xenografts have been the mainstay of cancer research for the past several decades. However, recent evidence suggests that therapeutic response is greatly influenced by the tumor cell microenvironment. Therefore, we have developed an ex vivo analysis of primary tumor specimens for drug development purposes.

기본 종양 및 / 또는 전임상에 대한 관련 전이 및 치료제의 임상 개발의 전 생체 치료 응답

The molecular analysis of established cancer cell lines has been the mainstay of cancer research for the past several decades. Cell culture provides both ...

Isolated Hepatic Perfusion as a Treatment for Liver Metastases of Uveal Melanoma

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic agent melphalan. Briefly, the procedure starts with the setup of a percutaneous veno-venous bypass from the femoral vein to the external jugular vein. Via a laparotomy, catheters are then inserted into the proper hepatic artery and the caval vein. The portal vein and the caval vein, both supra- and infrahepatically, are then clamped. The arterial and venous catheters are connected to a heart lung machine and the liver is perfused with melphalan (1 mg/kg body weight) for 60 min. This way it is possible to locally perfuse the liver with a high dose of a chemotherapeutic agent, without leakage to the systemic circulation.
In previous studies including patients with isolated liver metastases of uveal melanoma, an overall response rate of 33-100% and a median survival between 9 and 13 months, have been reported. The aim of this protocol is to give a clear description of how to perform the procedure and to discuss IHP as a treatment option for liver metastases of uveal melanoma.

Here, we present a protocol how to perform an isolated liver perfusion (IHP) with melphalan and we also discuss IHP as a treatment option for liver metastases of uveal melanoma.

포도막 흑색 종의 간전이에 대한 치료로 고립 간 관류

Isolated hepatic perfusion (IHP) is a procedure where the liver is surgically isolated and perfused with a high concentration of the chemotherapeutic ...

Percutaneous Hepatic Perfusion (PHP) with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn unresectable metastases into resectable ones. Some patients however, suffer from side effects or progression under systemic treatment. For patients with metastasized uveal melanoma there are no standard systemic therapy options. For patients without extrahepatic disease, isolated liver perfusion (IHP) may enable local disease control with limited systemic side effects. Previously, this was performed during open surgery with satisfying results, but morbidity and mortality related to the open procedure, prohibited a widespread application. Therefore, percutaneous hepatic perfusion (PHP) with simultaneous chemofiltration was developed. Besides decreasing morbidity and mortality, this procedure can be repeated, hopefully leading to a higher response rate and improved survival (by local control of disease). During PHP, catheters are placed in the proper hepatic artery, to infuse the chemotherapeutic agent, and in the inferior caval vein to aspirate the chemosaturated blood returning through the hepatic veins. The caval vein catheter is a double balloon catheter that prohibits leakage into the systemic circulation. The blood returning from the hepatic veins is aspirated through the catheter fenestrations and then perfused through an extra-corporeal filtration system. After filtration, the blood is returned to the patient by a third catheter in the right internal jugular vein. During PHP a high dose of melphalan is infused into the liver, which is toxic and would lead to life threatening complications when administered systemically. Because of the significant hemodynamic instability resulting from the combination of caval vein occlusion and chemofiltration, hemodynamic monitoring and hemodynamic support is of paramount importance during this complex procedure.

Percutaneous Hepatic Perfusion PHP with Melphalan as a Treatment for Unresectable Metastases Confined to the Liver

In this manuscript, we describe percutaneous isolated hepatic perfusion with simultaneous chemofiltration as treatment for unresectable liver metastases. This procedure is performed under general anaesthesia in the angiosuite by an experienced team, consisting of an interventional radiologist, a clinical perfusionist and anaesthesiologist.

간으로 밀폐 절제 불가능한 전이에 대한 치료로 멜 팔란과 경피적 간 관류 (PHP)

Unresectable liver metastases of colorectal cancer can be treated with systemic chemotherapy, aiming to limit the disease, extend survival or turn ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Myography 와이어를 사용 하 여 마우스 Mesenteric 동맥의 isometric 수축 측정

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

당뇨병 토끼에서의 뒷 사지의 전 임상 모델

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...