Name: intraductal delivery to the rabbit mammary gland
Uploaded: 2017-03-09T13:00:00.000+00:00
Duration: 6 min 30 s
Description: Watch this Scientific Journal Video about Intraductal Delivery to the Rabbit Mammary Gland at JoVE.com

The authors acknowledge support from a Translational Breast Cancer Research Grant (14-60-26-BROC to AB) from the Breast Cancer Research Foundation and the American Association for Cancer Research.

동물의 주제를 사용하는 절차는 텍사스 오스틴 대학의 기관 동물 관리 및 사용위원회에 의해 승인되었습니다. 1. 수술 전 준비 <ol><li> 각각의 토끼의 체중을 기록한다. 모든 임상 시험과 같이, 잠재적 인 독성을 평가하기 위해 정기적으로 동물의 무게를 모니터링한다. </li><li> 전에 토끼를 마취로, 시중에서 판매하는 초음파 젤 (30)의 500 XG에 스핀 50 ML 원뿔 관을 채우기; 원심 분리 완료시 겔에 가시적 인 거품이 없어야합니다. </li><li> 0.75 ㎎ / ㎏의 투여 량으로 근육 내 아세 프로 마진, 0.1 ㎎ / ㎏의 용량으로 피하 글리코을 관리하고. 생체 신호와 동작을 모니터링하면서 적용하려면 진정을 위해 15 ~ 20 분을 기다립니다. 참고 : 글리코은 서맥을 방지하고 호흡과 GI 분비물을 감소시키는 항 콜린성 에이전트입니다. 아세 프로 마진은 premedicati 역할을하는 진정제입니다마취에. </li><li> 케타민 35 ㎎ / ㎏과 kg의 자일 라진 피하 마취로 5 mg을 /를 관리 할 수 ​​있습니다. 그러나 운전자가 경험하게되고 케타민 3 밀리그램의 / kg 15 mg의 약물 투여 량을 감소, 더 빠르게 췌관 전달을 수행 할 수 / 자일 라진 피하의 kg; 이것은 마취 시간 및 마취에서 회복하는 동물에 필요한 시간을 단축한다. </li><li> 확인하고 심장 박동, SPO 2, 온도, 호흡, 및 점액 막 색 매 15 분을 기록. 두 눈에 눈 윤활제를 적용합니다. 참고 : 적절한 훈련을받은 및 관리 또는 마취를 모니터링해야 그 기관의 IACUC에 의해 승인 된 인원 만. 수의사 마취 모니터의 사용은 생체 신호를 수집하고 권장에 도움이 될 수 있습니다. 수의학 마취 모니터 그러나, 수동으로 동물에게 15 분마다 체크를 할 필요를 대체하지 않는다. f를 제조업체의 지침을 참조하십시오또는 수의학 모니터의 적절한 사용. </li><li> 부드러운 발가락 핀치에 의해 마취의 발병을 확인; 토끼는 계속하기 전에 비 응답해야합니다. </li><li> 조심 사타구니 유두의 제 3의 형태 및 제 4 쌍의 주변에 토끼의 꼬리 복부를 면도. </li><li> 제거 된 머리의 대부분으로, 면도 영역에 오버 - 더 - 카운터 제모 크림을 적용합니다. 따뜻한 물에 젖은 축축한 종이 타월을 사용하는 응용 프로그램 후 크림 10 분을 제거합니다. 참고 : 토끼 피부가 민감한 / 매우 깨지기입니다. 제모 크림은 더 이상 10 분 동안 사용되어야한다. 실제로, 5 분으로 &quot;테스트 지점&quot;을 시도하고 테스트 스폿이 기간이 필요한 더 긴 시간을 나타내는 경우에만 이상에두고 안전하다. </li><li> 주사 부위를 청소하기 위해 알코올을 적신 거즈 패드 영역을 닦습니다. </li><li> 순환되는 온수 담요와 흡수 패드 늘어서 V 자형 골에서의 배부에 토끼를 놓습니다. </li></ol> 2. 홍보조영제의 eparation <ol><li> 제조업체의 지시에 따라 타겟팅되지 않은 대조 시약을 재구성한다. 최대 피펫 아래로 부드럽게 혼합. 주 :이 프로토콜에서 사용되는 조영제 4 실온에서 안정 - 재구성 후 6 시간. 조심스럽게 각 검색 사이의 유리 병 바위. 주 : 덕트의 수에 의존 할 필요 용액의 부피가 주입된다. 솔루션은 성인 뉴질랜드 화이트 토끼 (Oryctolaguscuniculus) 중 하나 도관 트리를 채우기에 충분하다의 토끼는 유두 당 4 유관 구멍, 0.2 mL로있다. 이에 따라, 0.8 mL의 총 부피는 하나의 유선으로 전달 될 수있다. </li></ol> 3. 췌관 배달 <ol><li> 주입 할 수있는 적절한 유두를 찾습니다; 동물이 그것의 배부에 위치 할 때 쉽게 가시화 될 때 사타구니 유두의 3 번째 및 4 번째 쌍은, 사용하는 것이 좋습니다. </li><li> 로 멸균 0.9 % 식염수를 0.2 mL의 부하22 G 바늘 1 mL의 루어 잠금 투베르쿨린 주사기. 제대로 식염수를 주사기에 한 번 22 G 바늘 폐기 및 멸균 25 게이지 바늘로 교체합니다. 부드럽게 거즈 패드에 85 % 이소 프로필 알코올로 영역을 닦습니다. </li><li> 바늘 위로 동물의 몸에 주사기 병렬의 경사에 의해, 유두의 측면에 바늘의 베벨이 삽입 천천히 식염수 0.1-0.2 mL의 주입; 이것은 유관 개구의 더 나은 시각화 수 있습니다. </li><li> 로드 1 mL의 루어 잠금 투베르쿨린 주사기로 주입 용액 0.2 mL를. </li><li> 엄지와 검지 손가락으로 부드럽게 유두를 잡고 관내 주입을 위해 위치를 약간 들어 올려; 착용 안구 루페는 도관 개구부를 시각화에 도움이된다. </li><li> 유두의 상승 위치를 유지하면서 조심스럽게 25 G 뭉툭한 팁 바늘을 사용하여 관심있는 덕트를 cannulate. </li><li> 삽관 후, 부드럽게 무딘-t의 중심에있는 루어 락 주사기를 비틀제자리에 고정 될 때까지 IP를 주입 바늘. </li><li> 유두를 들어 올리고 빠르게 덕트 내에서의 유체 이동에 의한 손상을 최소화하기 위해 천천히 용액을 주입; 용액을 주입 할 때 짧은 시간에 저항이 있어야합니다. </li></ol> 4. 초음파 이미징 <ol><li> 관심 영역의 피부에 원심 분리 초음파 젤의 자유로운 금액을 적용합니다. 이러한 이미지 품질을 손상되므로, 겔에 기포가 없도록. </li><li> 6mm에 이미징 깊이를 설정합니다. 겔과 접촉하는 21 MHz의 진동자를 배치하고 B 모드에서 관심 영역을 스캔. 도관 개구 덕트에 걸쳐 포함하는 검사 영역의 조영제를 관찰한다. 주의 : 이러한 설정은 특정 음향 초음파 장치와 함께 사용하기 위해 개발되었다. 자세한 내용은 재료의 표를 참조하십시오. 이는 유방을 최적화하기 위해 송신 전력 및 다른 이미징 시스템의 촬상 깊이를 조정하는 것이 필요할 수있다그랜드 시각화. </li><li> 거즈 패드 동물의 피부에서 초음파 젤을 제거합니다. </li></ol> 5. 수술 후 케어 <ol><li> 유두 영역이나 주변 조직에 외상 흔적이 없어야하고, 유두 주변 영역에서 팽창하는 것으로 유방 지방 패드에 주입보다는 성공적인 췌관 주입 나타냄 주입 부위를 관찰한다. </li><li> 흉골 위치에 토끼를 놓습니다. 필요한 경우, 정맥 내 귀 가장자리 정맥 요힘빈 0.2 ㎎ / ㎏을 제공; 이 자일 라진의 효과를 반전하고, 동물이 마취로부터 더 빨리 복구하는 것을 허용 할 것이다. </li><li> 동물이 응답하는 경보이며, 흉골 위치에 유지 될 때까지 토끼에게 회복 기간 동안 15 분마다 모니터. 참고 :이 절차는 조직 손상이나 붓기가 발생하지 않아야합니다. 적색 또는 관찰 붓기 경우 동물이 경고와 수되면, 멜 록시 캄 0.1 ~ 0.2 ㎎ / ㎏ PO의 복용량을 관리구두 약을 복용합니다. 추가 지침에 대한 기관의 수의학 직원에게 문의하십시오. </li></ol>

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J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Addressing+Overdiagnosis+and+Overtreatment+in+Cancer:+A+Prescription+For+Change.">Addressing Overdiagnosis and Overtreatment in Cancer: A Prescription For Change.</a> Lancet Oncol. 15 (6), e234-e242 (2014).</li></ol>

The authors have nothing to disclose.

토끼 유방 선으로 관내 전달하는이 방법은 중요한 염료 및 치료를 포함하여, 초음파 대조 시약 및 기타 수용액에 사용될 수있다. 이전의 연구는 호르몬 17, 18, 19의 췌관 배달을 증명하고있다. 설치류 모델 핵산 (8)의 관내 전달, 화학 요법 제 6, 7, 나노 캐리어 <sup clas...

<html> 치료제의 관내 전달은 설치류 모델에서 초기 및 인간 실험 3, 4, 5, 6, 11, 12에서 검토되고있다. 공부의 최근 단계는 췌관 카보 플 라틴 또는 침습성 암 (2)의 치료를 위해 유방 절제술을 기다리고 여성의 췌관 페 길화 된 리포좀 독소루비신의 안전과 가능성을 보여 주었다. 관내 전달을 위해 이전 프로토콜은 마우스와 랫트 젖샘 6, 7, 8, 9에 대해 개발되어왔다. 연구 목적, 관내 종양 세포를 주사하고 암 유전자의 렌티 바이러스 벡터 전달에도 설치류 모델에서 수행 된REF &quot;&gt; 13, 14, 15, 16. 그러나 관내 전달 프로세스의 생체 내 모델에서 이상적 치료 화합물의 신규 클래스의 개발을 허용하고 임상 평가를 용이하게한다. 설치류와 인간 사이의 해부학 적 차이는 이들의 번역을 복잡했다 연구. 마우스와는 달리, 각각의 덕트는 별도의 젖꼭지에서 종료하는, 인간의 가슴은 5 ~ 9 독립적 인 유관 시스템, 유두에서 끝나는 별도의 개방 각으로 구성되어 있습니다. 토끼 유방 땀샘은 네 개의 독립 유관 시스템, 하나의 젖꼭지에 네 개의 구멍 중 하나를 통해 각각 개별적으로 접근 항구. 토끼 모델은 더 밀접하게 일치 인체 해부학 및 더 중요한 상황에서 관내 약물 전달 연구를 허용한다. 여기, 우리는 관내 배달을 평가하기 위해이 기술을 사용합니다. (A)의 공동 투여중요한 색소는 피부로 시각화를 허용하고있어서 간단하고 신속하게 확인을 제공한다. 일부 애플리케이션의 경우, 덕트 고해상도 매핑은 바람직 할 수있다. 우리는 비 대상 대조 시약 관내 전달을 통해 여기 덕트의 초음파 영상을위한 프로토콜을 제시한다.

<table><tbody><tr><td>MicroMarker non-targeted contrast reagent</td><td>VisualSonics</td><td>VS-11694</td><td></td></tr><tr><td>Luer Lock 1 mL Syringes</td><td>BD</td><td>309628</td><td></td></tr><tr><td>Glycopyrrolate 0.2 mg/mL</td><td>Wedgewood Compounding Pharmacy</td><td>GLYCOP-INJ013VC</td><td>6 month shelf life, supply may be limited.&amp;nbsp;</td></tr><tr><td>Atropine Sulfate 0.5 mg/mL</td><td>Animal Health International</td><td>15320764</td><td>If glycopyrrolate is unavailable. Not to be combined with glycopyrrolate.</td></tr><tr><td>Ketamine HCL 100 mg/mL</td><td>Animal Health International</td><td>21250699</td><td>&lt;a href=&quot;http://www.animalhealthinternational.com/&quot;&gt;http://www.animalhealthinternational.com/&lt;/a&gt;</td></tr><tr><td>Acepromazine 10 mg/mL</td><td>Animal Health International</td><td>17640541</td><td></td></tr><tr><td>Xylazine 20 mg/mL</td><td>Animal Health International</td><td>20101547</td><td></td></tr><tr><td>Yohimbine 0.2 mg/mL</td><td>Animal Health International</td><td>14588965</td><td></td></tr><tr><td>Hair Removing Cream</td><td>Veet</td><td></td><td>Sensitive skin solution. Available through local retailers.</td></tr><tr><td>Blunt tip infusion needles</td><td>Sai Infusion Technology</td><td>B14-50</td><td>&lt;a href=&quot;http://www.sai-infusion.com/collections/blunt-needles&quot;&gt;http://www.sai-infusion.com/collections/blunt-needles&lt;/a&gt;</td></tr><tr><td>Veterinary Pulse Oximeter</td><td>EdanUSA</td><td>VE-H100B</td><td>&lt;a href=&quot;http://www.edanusa.com/Product/VE-H100B-Veterinary-Pulse-Oximeter.html&quot;&gt;http://www.edanusa.com/Product/VE-H100B-Veterinary-Pulse-Oximeter.html&lt;/a&gt;</td></tr><tr><td>Warm Water Pump</td><td>Gaymar</td><td>TP700</td><td></td></tr><tr><td>Warm Water Blanket</td><td>Animal Health International</td><td>21232696</td><td>Maxi-Therm Lite Warming Pads</td></tr><tr><td>Ultrasound system</td><td>VisualSonics</td><td>Vevo 2100</td><td></td></tr></tbody></table>

intraductal delivery to the rabbit mammary gland

유방암 췌관 국소 치료는 종양에 대한 효율적인 전달 및 감소 된 독성 및 부작용이 1, 2, 3, 4, 5, 6, 7을 포함한 잠재적 인 이점을 제공한다. 이러한 치료가 더 널리 적용 할 수 있습니다 전에 그러나, 몇 가지 과제가 남아있다. 적절한 동물 모델의 개발과 췌관 치료제의 유효성 검사는 환자 췌관 치료 전략의 개발을 용이하게한다. 마우스 유방 선 널리 유선 종양의 발달 및 모델 시스템으로 사용되었지만, 해부학 인간 동맥 구별된다. 더 큰 동물 모델은 같은 토끼로, 유선 구조 및 췌관 치료 개발을위한 더 나은 모델이 될 수 있습니다. C에서마우스에 ontrast,하는 열 유관 나무 공간적으로 분리 된 유두의 각 종단은 토끼 유방 동맥은 더 밀접하게 한 젖꼭지에 별도의 구멍을 통해 그 출구 여러 중복 유관 시스템과, 인간의 동맥과 유사한 신체의 축을 따라 배포됩니다. 여기서는 직접 유선 토끼 덕트로 고해상도 초음파 영상과 함께 전달 자체의 시각화를위한 시약의 전달을위한 최소 침습적 방법을 제시한다.

여기서는 토끼 유방 덕트 대조적 시약의 관내 전달은 조직 (도 2)에 충격없이 달성 될 수 있다는 것을 보여준다. 토끼에서, 네 개의 별도의 유관 시스템은 하나의 젖꼭지에 수렴 따라서 액세스 할 수 있으며,이 방법을 사용하여 개별적으로 몇 군데. 개인 유관 구멍이 쉽게 나타나는; 도 2b에서 유관 덕트에 인접한 제 2 도관 개구를 표시 화살촉을 확?...

Watch this Scientific Journal Video about Intraductal Delivery to the Rabbit Mammary Gland at JoVE.com

Intraductal Delivery to the Rabbit Mammary Gland

Here, we describe a technique for the localized delivery of reagents to the rabbit mammary gland via an intraductal injection. In addition, we describe a protocol for visualization and the confirmation of delivery by high-resolution ultrasound imaging of contrast agents.

토끼 유선으로 관내 배달

Localized intraductal treatments for breast cancer offer potential advantages, including efficient delivery to the tumor and reduced systemic toxicity and ...

intraductal-delivery-to-the-rabbit-mammary-gland

Research

JoVE Journal

Bioengineering

9.3K 조회수.  The University of Texas at Austin. Here, we describe a technique for the localized delivery of reagents to the rabbit mammary gland via an intraductal injection. In addition, we describe a protocol for visualization and the confirmation of delivery by high-resolution ultrasound imaging of contrast agents. 

비디오: Intraductal Delivery to the Rabbit Mammary Gland

Minimally Invasive Embryo Transfer and Embryo Vitrification at the Optimal Embryo Stage in Rabbit Model

Assisted reproductive techniques (ARTs), such as in vitro embryo culture or embryo cryopreservation, affect natural development patterns with perinatal and postnatal consequences. To ensure the innocuousness of ART applications, studies in animal models are necessary. In addition, as a last step, embryo development studies require evaluation of their capacity to develop full-term healthy offspring. Here, embryo transfer to the uterus is indispensable to perform any ARTs-related experiment.
The rabbit has been used as a model organism to study mammalian reproduction for over a century. In addition to its phylogenetic proximity to the human species and its small size and low maintenance cost, it has important reproductive characteristics such as induced ovulation, a chronology of early embryonic development similar to humans and a short gestation that allow us to study the consequences of ART application easily. Moreover, ARTs (such as intracytoplasmic sperm injection, embryo culture, or cryopreservation) are applied with suitable efficiency in this species.
Using the laparoscopic embryo transfer technique and the cryopreservation protocol presented in this article, we describe 1) how to transfer embryos through an easy, minimally invasive technique and 2) an effective protocol for long-term storage of rabbit embryos to provide time-flexible logistical capacities and the ability to transport the sample. The outcomes obtained after transferring rabbit embryos at different developmental stages indicate that morula is the ideal stage for rabbit embryo recovery and transfer. Thus, an oviductal embryo transfer is required, justifying the surgical procedure. Furthermore, rabbit morulae are successfully vitrified and laparoscopically transferred, proving the effectiveness of the described techniques.

Assisted reproductive techniques (ARTs) are in continuous evaluation to improve outcomes and reduce the associated risks. This manuscript describes a minimally invasive embryo transfer procedure with an efficient cryopreservation protocol that allows the use of rabbits as an ideal animal model of human reproduction.

토끼 모형에 있는 최적 배아 단계에서 최소 침 습 배아 이식 및 배아 확산

Assisted reproductive techniques (ARTs), such as in vitro embryo culture or embryo cryopreservation, affect natural development patterns with perinatal ...

연구

발생학

Intraductal Delivery and X-ray Visualization of Ethanol-Based Ablative Solution for Prevention and Local Treatment of Breast Cancer in Mouse Models

Breast cancer is the most prevalent cancer and the second-leading cause of cancer-related death for women in the USA. For high-risk women, prophylactic mastectomy is the most effective primary prevention strategy. Prophylactic mastectomy is an aggressive surgical procedure that completely removes the mammary epithelial cells from which breast cancer arises along with the surrounding tissue. We seek to develop a minimally invasive intraductal procedure as an alternative to prophylactic mastectomy to locally ablate the mammary epithelial cells before they can become malignant. We and others have developed an intraductal delivery procedure to reach and treat these epithelial cells in rodent models of breast cancer. While the mouse mammary gland with a single non-anastomosed ductal tree opening at the nipple has a much less complex and tortuous architecture than the human breast, chemically induced and genetically engineered mouse models of breast cancer are valuable to produce proof-of-concept studies of new preventative strategies. Here, we describe a procedure for intraductal delivery of an ethanol-based ablative solution containing micro-CT/X-ray tantalum-based contrast agent within the mouse mammary ductal tree for the therapeutic purpose of primary prevention of breast cancer. Intraductal delivery of aqueous reagents (e.g., cytotoxic compounds, siRNAs, AdCre) has been previously described in mouse models. Thus, we focus our protocol description on methodological modifications and unique experimental considerations for optimizing delivery of ethanol, for minimizing local and systemic side effects of ethanol administration, and for in vivo visualization of ductal tree filling via micro-CT/fluoroscopy imaging. Visualization of the ductal tree immediately after injection of a contrast-containing solution allows for confirmation of complete filling or unsuccessful outcomes such as underfilling or overfilling. This procedure can be applied for delivery and imaging of other ablative compounds aimed at either preventing tumor formation or locally treating early-stage tumors accessible via the ductal tree.

A method of intraductal injection of reagents for an ethanol-based ablative solution to the mouse mammary ductal tree for in vivo imaging and breast cancer prevention is described. Injection directly into the nipple opening allows for targeting mammary epithelial cells with minimal collateral tissue damage.

마우스 모델에서 유방암의 예방 및 지역 치료를 위한 에탄올 기반 담약 솔루션의 내배송 및 X선 시각화

Breast cancer is the most prevalent cancer and the second-leading cause of cancer-related death for women in the USA. For high-risk women, prophylactic ...

암 연구학

X-Ray Visualization of Intraductal Ethanol-Based Ablative Treatment for Prevention of Breast Cancer in Rat Models

There are still a limited number of primary interventions for prevention of breast cancer. For women at a high risk of developing breast cancer, the most effective intervention is prophylactic mastectomy. This is a drastic surgical procedure in which the mammary epithelial cells that can give rise to breast cancer are completely removed along with the surrounding tissue. The goal of this protocol is to demonstrate the feasibility of a minimally invasive intraductal procedure that could become a new primary intervention for breast cancer prevention. This local procedure would preferentially ablate mammary epithelial cells before they can become malignant. Intraductal methods to deliver solutions directly to these epithelial cells in rodent models of breast cancer have been developed at Michigan State University and elsewhere. The rat mammary gland consists of a single ductal tree that has a simpler and more linear architecture compared to the human breast. However, chemically induced rat models of breast cancer offer valuable tools for proof-of-concept studies of new preventive interventions and scalability from mouse models to humans. Here, a procedure for intraductal delivery of an ethanol-based ablative solution containing tantalum oxide nanoparticles as X-ray contrast agent and ethyl cellulose as gelling agent into the rat mammary ductal tree is described. Delivery of aqueous reagents (e.g., cytotoxic compounds, siRNAs, AdCre) by intraductal injection has been described previously in mouse and rat models. This protocol description emphasizes methodological changes and steps that pertain uniquely to delivering an ablative solution, formulation consideration to minimize local and systemic side effects of the ablative solution, and X-ray imaging for in vivo assessment of ductal tree filling. Fluoroscopy and micro-CT techniques enable to determine the success of ablative solution delivery and the extent of ductal tree filling thanks to compatibility with the tantalum-containing contrast agent.

A procedure for the delivery of a chemical ablative solution to the rat mammary ductal tree for image-guided preventive treatment of breast cancer is described. Mammary epithelial cells can be targeted with minimal collateral tissue damage through cannulation directly into the nipple opening and intraductal infusion of a 70% ethanol-based ablative solution.

쥐 모델에서 유방암 예방을 위한 관내 에탄올 기반 절제 치료의 X선 시각화

There are still a limited number of primary interventions for prevention of breast cancer. For women at a high risk of developing breast cancer, the most ...

In Vivo Gene Delivery into Mouse Mammary Epithelial Cells Through Mammary Intraductal Injection

Mouse mammary glands comprise ductal trees, which are lined by epithelial cells and have one opening at the tip of each nipple. The epithelial cells play a major role in mammary gland function and are the origin of most mammary tumors. Introducing genes of interest into mouse mammary epithelial cells is a critical step in evaluating gene function in epithelial cells and generating mouse mammary tumor models. This goal can be accomplished through the intraductal injection of a viral vector carrying the genes of interest into the mouse mammary ductal tree. The injected virus subsequently infects mammary epithelial cells, bringing in the genes of interest. The viral vector can be lentiviral, retroviral, adenoviral, or adenovirus-associated viral (AAV). This study demonstrates how a gene of interest is delivered into mammary epithelial cells through mouse mammary intraductal injection of a viral vector. A lentivirus carrying GFP is used to show stable expression of a delivered gene, and a retrovirus carrying Erbb2 (HER2/Neu) is used to demonstrate oncogene-induced atypical hyperplastic lesions and mammary tumors.

In Vivo Gene Delivery into Mouse Mammary Epithelial Cells Through Mammary Intraductal Injection

The present protocol describes intraductal injection of viral vectors via the teat to deliver genes of interest into the mammary epithelial cells.

생체 내 유방 관내 주입을 통한 마우스 유방 상피 세포로의 유전자 전달

Mouse mammary glands comprise ductal trees, which are lined by epithelial cells and have one opening at the tip of each nipple. The epithelial cells play ...

Intranasal Immunization and Milk Collection in Studies of Maternal Immunization in New Zealand White Rabbits (Oryctolagus cuniculus)

Due to similarities in placentation and antibody transfer with humans, rabbits are an excellent model of maternal immunization. Additional advantages of this research model are the ease of breeding and sample collection, relatively short gestation period, and large litter sizes. Commonly assessed routes of immunization include subcutaneous, intramuscular, intranasal, and intradermal. Nonterminal sample collection for the chronological detection of the immunologic responses to these immunizations include the collection of blood, from both dams and kits, and milk from the lactating does. In this article, we will demonstrate techniques our lab has utilized in studies of maternal immunization in New Zealand White rabbits (Oryctolagus cuniculus), including intranasal immunization and milk collection.

Intranasal Immunization and Milk Collection in Studies of Maternal Immunization in New Zealand White Rabbits Oryctolagus cuniculus

This article describes and demonstrates the administration of intranasal vaccines and the collection of milk from lactating rabbits (Oryctolagus cuniculus) as a means to assess mucosal immunity in a translationally appropriate model of maternal immunization.

뉴질랜드 흰 토끼의 모성 예방 접종 연구에서 비강 내 예방 접종 및 우유 수집(Oryctolagus cuniculus)

Due to similarities in placentation and antibody transfer with humans, rabbits are an excellent model of maternal immunization. Additional advantages of ...

면역학 및 감염병학

Intraductal Injection for Localized Drug Delivery to the Mouse Mammary Gland

Herein we describe a protocol to deliver various reagents to the mouse mammary gland via intraductal injections. Localized drug delivery and knock-down of genes within the mammary epithelium has been difficult to achieve due to the lack of appropriate targeting molecules that are independent of developmental stages such as pregnancy and lactation. Herein, we describe a technique for localized delivery of reagents to the mammary gland at any stage in adulthood via intraductal injection into the nipples of mice. The injections can be performed on live mice, under anesthesia, and allow for a non-invasive and localized drug delivery to the mammary gland. Furthermore, the injections can be repeated over several months without damaging the nipple. Vital dyes such as Evans Blue are very helpful to learn the technique. Upon intraductal injection of the blue dye, the entire ductal tree becomes visible to the eye. Furthermore, fluorescently labeled reagents also allow for visualization and distribution within the mammary gland. This technique is adaptable for a variety of compounds including siRNA, chemotherapeutic agents, and small molecules.

A protocol for the non-invasive intraductal delivery of aqueous reagents to the mouse mammary gland is described. The method takes advantage of localized injection into the nipples of mammary glands targeting mammary ducts specifically. This technique is adaptable for a variety of compounds including siRNA, chemotherapeutic agents and small molecules.

마우스 유선 화 된 약물 전달을위한 췌관 주입

Herein we describe a protocol to deliver various reagents to the mouse mammary gland via intraductal injections. Localized drug delivery and knock-down of ...

생물학

Simplifying Transcutaneous Intratracheal Drug Delivery in the Newborn Preterm Rabbit

Intratracheal (IT) drug delivery allows the direct delivery of pharmaceutical substances to the lung, maximizing potential pulmonary benefit and minimizing systemic drug exposure. The transcutaneous technique is simple and allows for the IT delivery of substances to the lung of prematurely born rabbits shortly after birth. Newborn pups are anesthetized with inhaled Isoflurane before being placed in a supine position with the neck extended. The larynx is identified and stabilized before transcutaneous placement of a 26-gauge (G) catheter into the trachea. Following catheterization of the trachea, a 30 G blunt needle attached to a Hamilton syringe is introduced into the IT catheter and is used for delivering a precise volume into the trachea during spontaneous respiration. After the IT injection is completed, the needle and catheter are withdrawn, and the pup is allowed to recover from anesthesia. Transcutaneous IT injection delivers a large proportion of the injected substance to the lung, with the majority remaining in the lung 3 hours after the intervention. The injections are well tolerated from the day of birth and can be repeated for multiple consecutive days without influencing survival. This technique can be used to investigate the effect of pharmaceutical agents on lung development and in the prevention of neonatal lung injury in preterm rabbits.

Transcutaneous intratracheal injection allows for effective intrapulmonary drug delivery during spontaneous respiration. Single and multiple injections are well tolerated with no effect on survival. The technique is simple to perform and can examine the effect of substances on lung development and the prevention of lung injury in newborn rabbits.

Intratracheal (IT) drug delivery allows the direct delivery of pharmaceutical substances to the lung, maximizing potential pulmonary benefit and ...

의학

Pregnancy and Nursing Management for Embryo-Transferred and Genetically Modified Rabbits

With the advancement of scientific research, the demand for gene-edited rabbit models is increasing. However, there are limited pregnancy and feeding management systems for gene-edited rabbits, leading to low survival rates among gene-edited rabbits prepared by many inexperienced researchers. Therefore, proper guidance is essential. This article summarizes the pregnancy and feeding practices for genetically modified rabbits developed in the author's laboratory and outlines a set of fundamental processes. These include pregnancy diagnosis, antenatal care, midwifery, assisted breastfeeding, weaning, and other procedures, along with the rescue and care of weak newborn rabbits. Compared to the traditional natural childbirth and nurturing methods used in rabbit farms, this approach involves more refined management, requiring additional time and effort but significantly increasing the survival rate of suckling rabbits. The methods described in this article are suitable for most laboratory breeding scenarios involving gene-edited or embryo-transferred rabbits and provide a straightforward and effective reference for other researchers.

This protocol describes a pregnancy and feeding management technique for embryo-transferred and genetically modified rabbits, aimed at reducing newborn rabbit mortality and enhancing the preparation efficiency of gene-edited rabbits.

With the advancement of scientific research, the demand for gene-edited rabbit models is increasing. However, there are limited pregnancy and feeding ...

Intraductal Injection: Delivering Injection Mix into the Ducts of the Mouse Mammary Gland

Source: Xiang, D., et al. <a href="https://www.jove.com/v/59502/modeling-breast-cancer-via-an-intraductal-injection-cre-expressing">Modeling Breast Cancer via an Intraductal Injection of Cre-expressing Adenovirus into the Mouse Mammary Gland.</a> J. Vis. Exp (2019).
This video describes an intraductal injection technique where we deliver injection mix into the ducts of mouse mammary glands.

Intraductal Injection: 마우스 유선의 관으로 주입 혼합물을 전달하기

Source: Xiang, D., et al. Modeling Breast Cancer via an Intraductal Injection of Cre-expressing Adenovirus into the Mouse Mammary Gland. J. Vis. Exp ...

JoVE Encyclopedia of Experiments

Breast Cancer

Procedures and techniques

Rabbit Intraductal Injection: Localized Delivery of Solution of Interest into the Rabbit Mammary Gland

Source: Clark et. al., <a href="https://www.jove.com/v/55209/intraductal-delivery-to-the-rabbit-mammary-gland">Intraductal Delivery to the Rabbit Mammary Gland.</a> J. Vis. Exp. (2017).
In this video we describe a technique of administering a solution of interest into the mammary duct of a rabbit and confirm the delivery by high-resolution ultrasound imaging of contrast agents.

Rabbit Intraductal Injection: Rabbit Mammary Gland로의 관심 용액의 국부적 전달

Source: Clark et. al., Intraductal Delivery to the Rabbit Mammary Gland. J. Vis. Exp. (2017).
In this video we describe a technique of administering a ...