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Medicina

自动子宫组织移植手术诱发子宫内膜异位症小鼠模型

Published: January 06, 2012
doi:
10.3791/3396
, ,
1Obstetrics, Gynecology and Women’s Health and Division of Biological Sciences,University of Missouri, 2Obstetrics, Gynecology and Women’s Health and Animal Sciences,University of Missouri

Summary

子宫内膜异位症手术诱导小鼠和大鼠子宫组织的自动移植到肠系膜动脉级联的描述。

Abstract

子宫内膜异位症是一种慢性，痛苦的疾病，其病因尚不清楚。此外，子宫内膜异位症的治疗，可以要求腹腔镜切除病灶和/或慢性疼痛和不孕症状的药品管理。已与子宫内膜异位症相关的成本估计为每年220亿美元，在美国的1 。为了进一步了解我们这个谜一般的疾病的基本机制的，动物模型已被采用。灵长类动物的自发发展子宫内膜异位症，因此灵长类动物模型最接近妇女疾病。然而，啮齿动物模型更具成本效益和随时可用2。我们在这里描述的模型，涉及到肠系膜（图1）自体子宫组织转移，并首次在大鼠 3，后来转移到鼠标的4 。手术引起的子宫内膜异位症的自体啮齿动物模型的目标是模仿在女性的疾病。我们和其他人曾在小鼠或大鼠镜观察到，在妇女疾病5,6子宫内膜异位病灶观察表明，改变基因的表达模式。鼠标在执行手术的优点之一是，大量的转基因小鼠品系可以帮助研究人员确定的具体组成部分子宫内膜异位症的建立和发展中的重要作用。另一种模式，即切除子宫内膜碎片是免疫功能低下小鼠的腹膜也被广泛使用，但受限于缺乏正常的免疫系统被认为是在子宫内膜异位症2,7的重要。重要的是，手术引起的子宫内膜异位症的小鼠模型是一个通用的模型已被用于研究免疫系统8，9,10和 11,12激素环境因素如何影响子宫内膜异位症，以及子宫内膜异位症的影响，对fertility 13和痛苦14。

Protocol

1。规划活畜手术确保相应的审批已收到与实验动物工作。 订购小鼠，并允许至少一个星期驯化新的环境。 雌性小鼠安置在接触到男性费洛蒙的情况下，可以停止循环，这种现象被称为惠滕效果15,16。为了保持骑自行车转移小鼠尿液浸泡过的男性床上用品的女性笼，每5天。另外，如果使用开顶式网箱，地方两个男性笼之间的女性笼保持经常骑自行车的女性。 ?…

Discussion

有几个关键参数，应当指出，在执行子宫内膜异位症手术诱导小鼠。首先，子宫内膜异位症是一种雌激素依赖性疾病，应在完整的动物，或者在去卵巢动物辅以雌激素 20进行这种手术。二，缝合子宫内膜活检动脉级联必须格外小心。我们发现，只有两个有一个相对宽松的结抛出保持活检，同时防止结扎肠和随后的组织坏死和动物死亡的血液供应。我们强烈建议练上几个小鼠前实际实验，?…

Declarações

The authors have nothing to disclose.

Acknowledgements

克里斯Kassotis这个手稿的严格审查和奥黛丽贝利和斯科特Korte博士，约瑟夫比曼，艾莉森Curfman，保罗甘，布里奇特Neibreggue，雅各Redel，艾米施罗德，迈亚斯坦伯格和Stacey Winkeler特别感谢他们在优化援助这种模式在我们的实验室。提供资助的临床Biodetectives培训资助（NIH T90的）（KEP），密苏里州的生命科学大学本科生研究机会计划研究理事会，穆，穆研究委员会的赠款和NIH R21HD056441（SCN）。

Materials

Name of the reagent/equipment Company Catalogue number
Wax pencil Fisher NC9954135
Glass slide Fisher 12-550-433
Eyedropper Fisher S79383
Standard light microscope for evaluating vaginal cytology smears    
Buprenorphine HCL c3 (CARJET) 10X1ml Butler Animal Health Supply 022891
Sterile phosphate buffered saline (PBS) Gibco 14040-117
10,000U/ml Penicillin, 10,000μg/ml Streptomycin in 0.85% NaCl Hyclone SV30010
Isoflurane Abbott Animal Health 05260-05
Isoflurane non-rebreathing anesthetic system    
Recirculating hot water heating pad    
30 ml syringe sheath Fisher 14-823-16G
Powder free sterile gloves Fisherbrand 19020558
Ophthalmic ointment Major Pharmaceuticals 10033691
Small electrical clippers Wahl 9861-600
Chlorhexidine scrub Fisher NC9863042
70% Ethanol    
Polylined sterile field Busse Hospital Disposables 696
Size 3 scalpel Fisher 22-079-657
Number 10 scalpel blades Fisher 22-079-681
Small surgical scissors Roboz RS-5850
Small serrated semi-curved forceps Roboz RS-5135
5-0 black braided silk suture Ethicon K870H
Sterilized pyrex glass Petri dishes Corning 70160-101
2 mm biopsy punch Miltex 33-31
Sterile gauze Kendall 1806
6-0 black monofilament ethilon nylon suture Ethicon 697G
Needle drivers (optional) World Precision Instruments 500023
5-0 undyed braided coated vicryl suture Ethicon J490G
9mm Autoclip wound clips Becton Dickinson 427631
Autoclip applier & remover Becton Dickinson 427630
23G needle Becton Dickinson 305193
1cc syringe Becton Dickson 301025
5X magnifying glass stand (optional) Fisher 14-648-23
10% Buffered formalin Fisher SF100-4
Calipers Roboz RS-6466
Processing/embedding cassettes Fisher 15-197-700A
Biopsy foam pads Fisher 22-038-222
RNAqueous RNA isolation kit Ambion AM1912
Liquid nitrogen    
Snap cap microcentrifuge flat top tube Fisher 02-681-240
Ketamine (optional) Simga K4138
Domitor (medetomidine hydrochloride) (optional) Tocris 2023
Antisedan (atipamezole) (optional) Sigma A9611

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Tags

EndometriosisMouse ModelSurgically-inducedAuto-transplantationChronic DiseaseEtiologyLaparoscopic RemovalPharmaceutical ManagementPainInfertility SymptomsCostAnimal ModelsPrimate ModelsRodent ModelsAutologous TransferIntestinal MesenteryGene Expression PatternTransgenic Mouse StrainsEstablishmentGrowth
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Pelch, K. E., Sharpe-Timms, K. L., Nagel, S. C. Mouse Model of Surgically-induced Endometriosis by Auto-transplantation of Uterine Tissue. J. Vis. Exp. (59), e3396, doi:10.3791/3396 (2012).
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