慢性病仪表非麻醉的胎羊模型被用于研究在健康和疾病的人类胎儿发育,因为它允许外科安置和维护导管和电极,反复血液采样,物质喷射,记录的生物电活动的,以及在体内成像 。我们描述了建立这种模式所需的程序。
The chronically instrumented pregnant sheep has been used as a model of human fetal development and responses to pathophysiologic stimuli such as endotoxins, bacteria, umbilical cord occlusions, hypoxia and various pharmacological treatments. The life-saving clinical practices of glucocorticoid treatment in fetuses at risk of premature birth and the therapeutic hypothermia have been developed in this model. This is due to the unique amenability of the non-anesthetized fetal sheep to the surgical placement and maintenance of catheters and electrodes, allowing repetitive blood sampling, substance injection, recording of bioelectrical activity, application of electric stimulation and in vivo organ imaging. Here we describe the surgical instrumentation procedure required to achieve a stable chronically instrumented non-anesthetized fetal sheep model including characterization of the post-operative recovery from blood gas, metabolic and inflammation standpoints.
对正常和损害怀孕研究各种动物模型的存在,包括实验室啮齿类,非人类的灵长类动物和家养反刍动物。1,2,3,4,5长期仪器妊娠母羊已被广泛使用50年人类胎儿发育和答复的模型的病理生理刺激如脂多糖(LPS)。6-10以下的LPS暴露病变模仿正是由于这两个物种的一个类似的成熟信息见于早产儿脑室周围白质软化,这是。 11,12
其他妊娠并发症也已经研究的很详细,如发现产前糖皮质激素促进肺的发育13-15和了解胎儿宫内发育迟缓(IUGR)对胎儿16,17的影响。
广泛使用的胎羊模型的是由于uniq的非麻醉胎羊到外科位置和维护导管和电极的UE顺从,允许重复的血液采样,记录的生物电活动的,应用电刺激和体内脑成像。18遥测也是可能的,虽然不经常使用的但由于较高的复杂性来设置,以及初始和维护成本。19
此外,胎羊模型是非常通用的仪器的许多变化是可能根据目标的措施。例如,有可能在数天来记录到周实时多元信号如胎儿呼吸运动,电脑活动,心血管反应,心电图,使用流量探针或微球体等由于局部血流的范围器官这种多功能性,已经进行了包括CARDI的发展广泛的研究ovascular系统20,21,下丘脑-垂体-肾上腺轴(HPA)22,大脑发育23和睡眠状态的发展,特别是24,缺氧的影响/窒息25,低温治疗26,炎症6-11,无论是27的组合,糖皮质激素28,29,抗抑郁药30,支气管肺发育不良(BPD)31,32,胎儿编程33,34,35,36,37,38,39或新颖的胎儿监护模式开发之前和分娩期间的名字,但调查的几个领域。40,41,42,43
提出的方法的总体目标是展示这种多功能车型的基本实现。其允许建立多种研究胎儿生理和病理生理上一体化,器官,细胞和分子水平的急性和慢性实验方案。
麻醉及手术过程中呈现所需要的建立动物模型,为研究胎儿生理和病理生理:长期仪器非麻醉的胎羊。
该协议中的四个关键步骤应该强调。首先,通过母亲侧面传递导管和电极:重要的是,这是在一次以避免任何内部器官损伤。第二,固定之前外缝合胎儿uterotomy运行地点:这是至关重要的,以防止或尽量减少羊水和前子宫闭合羊膜随后缝合的损失。三,动脉插管:胎羊动脉?…
The authors have nothing to disclose.
authors gratefully acknowledge funding support from the Molly Towell Perinatal Research Foundation, Canadian Institutes of Health Research (CIHR), and Fonds de Recherche du Québec – Santé (FRQS) (to MGF) and CIHR-Quebec Training Network in Perinatal Research (QTNPR) (to LDD).
The authors wish to thank Esther Simard, Marco Bosa, Carl Bernard and Carmen Movila for technical assistance.
ACE Light source | Schott-Fostec | A20500 | |
Dissecting scissors | Fine Science Tools | 14060 – 11 | |
Angled dissecting scissors | Fine Science Tools | 15006 – 09 | |
Scalpel handle | Fine Science Tools | 10003 – 12 | alternating dissecting tool |
Curved scalpel blades #12 | Fine Science Tools | 10012 – 00 | alternating dissecting tool |
Bone scissors | Fine Science Tools | 16044 – 10 | |
S & T suture tying forceps | Fine Science Tools | 00272 – 13 | |
Dumont SS forceps – angled | Fine Science Tools | 11203 – 25 | |
Braided silk suture size 6-0 | Teleflex Medical | 07 – 30 – 10 | |
Medical Tape | transpore | 3M | |
Ketamine hydrochloride 100 mg/ml | Hospira | NDC 0409 – 2051 – 05 | Final Does is 80 mg/kg |
Tranqui Ved Injection (xylazine 100 mg/ml) | Vecdo | NDC 50989 – 234 – 11 | Final Does is 10 mg/kg |
Reactive orange 14 | Sigma – Aldrich | R – 8254 | |
Ringers Solution Components | Solution is gas equilibrated with 95% O2 and 5% Co2, final pH 7.4 | ||
Sodium chloride | Sigma – Aldrich | S7653 | Final Concentration: 118 mM |
Potassium chloride | Fisher Scientific | P217 – 3 | Final Concentration: 4.7 mM |
Calcium chloride dihydrate | Fisher Scientific | C79 – 500 | Final Concentration: 2.5 mM |
Potassium phosphate monobasic | Fisher Scientific | P -285 | Final Concentration: 1.2 mM |
Magnesium sulfate | J.T. Baker | Jan-00 | Final Concentration: 0.57 mM |
4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES) | Fisher Scientific | BP 310 – 500 | Final Concentration: 5.95 g/L |
Glucose | Sigma – Aldrich | G8270 | Final Concentration: 5.5 mM |
LifeWindow | Digicare Biomedical Technology | ||
CED bioamplifier and ADC units | Cambridge Electronic Design Limited, Unit 4, Science Park, Milton Road, Cambridge CB4 0FE ENGLAND. |
Bioamp: 1902; ADC: micro1401; Data acquisition software: Spike 2, V7.13 | |
Neurolog analog signal bioamplifier | Digitimer Ltd 37 Hydeway Welwyn Garden City Hertfordshire, AL7 3BE, England |
NL108A | |
ABL800Flex | Radiometer Canada; 200 Aberdeen Dr, London, ON N5V 4N2 | ||
Eppendorf 5804R | Eppendorf Canada; 2810 Argentia Road, #2 Mississauga, Ontario, L5N 8L2 |
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Arrow Jugular Catheterization Set | Arrow International, Inc., 2400 Bernville Road, Reading, PA 19605 USA | ||
Atravet | 10 mg/mL | ||
Diazepam | 5mg/mL | ||
Ketamine | Ketalar | 100 mg/mL | |
Propofol | 10 mg/mL | ||
SurgiVeT | Endotracheal Tubes; Smiths Medical ASD, Inc. St. Paul, MN 55112, USA | ||
Cook Airway Exchange Catheter with RAPI-FIT Adapters | Cook Critical Care 750, Bloomington IN 47402-0489 USA | ||
Dispomed Ventilator | Dispomed Ltd., 745 Nazaire-Laurin, Joliette, Quebec J6E 0L6 | ||
BD Insyte-W | Becton Dickinson, Infusion Therapy Systems Inc., 9450 S State St, Sandy Utah 84070 USA | 22 to 20 G; 1 in [0.9 x 25 mm] to 1.16 in [1.1 x 30 mm] | |
Edwards Lifesciences Ref: PX272 Pressure monitoring kit with TruWave Disposable Pressure | |||
LifeWindow LW6000 | Digicare Biomedical Technology 107 Commerce Road, Boynton Beach, FL 33426-9365 USA | ||
Gaymar | |||
Babcock | |||
Polyvinyl catheters | SCI (Scientific Commodities Inc.) | 2 meters | |
2-0 Vicryl | |||
Castroviejo scissors | |||
electrocardiogram (ECG) | LIFYY, Metrofunk Kabel-Union, Berlin, Germany | four copper electrodes in single sheath, 2 meters | |
2-O Vicryl | |||
3-0 Vicryl | |||
PDS II USP | |||
Trimethoprim sulfadoxine | |||
Ampicillin | |||
Stopcock | Argon Medical, Cat 041220001A | Double 4-way Stopcock with male luer lock | |
Needles | Tyco Healthcare 8881202389 | Monoject aluminum hub blunt needles, 22Gx, 0.7mmx 38.1mm: for fetal arterial and venous catheters | |
Needles | Tyco Healthcare 8881202322 | Monoject aluminum hub blunt needles, 16Gx, 1.6mmx38.1mm: for amniotic catheters |