أولا وقبل كل شيء، نشكر بصدق الأمهات اللاتي شاركن في هذا المشروع. بعد ذلك، نعترف بجميع الموظفين الذين ساعدوا ويسهلوا إجراءات أخذ العينات، طبيب التخدير، طبيب التخدير ممرضة والممرضات الجراحية. ولم يكن ممكنا تحقيق هذا المشروع دون تمويل من هيئة الصحة الإقليمية في جنوب شرق النرويج والوحدة الاستشارية النرويجية المعنية بصحة المرأة، وجامعة أوسلو، والتمويل المحلي المقدم من مستشفى جامعة أوسلو.

<ol>
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الكتاب ليس لديهم ما يكشف.

<html> طريقة أخذ العينات من 4 أوعية للمشيمة ذات صلة بثلاثة أغراض رئيسية. أولا، يمكن استخدامه لدراسة كيفية تناول المواد المحددة من قبل المشيمة على الجانب الأمهات وربما نقلها إلى الدورة الدموية السري والجنين، كما يتضح من الجلوكوز والدراسات الأحماض الأمينية لدينا. ثانيا، ...

وفقا للمعاهد الوطنية للصحة، الولايات المتحدة الأمريكية، المشيمة هو الجهاز الأقل تفهما في جسم الإنسان 1 ، 2 ، 3 . فمن الصعب الوصول إلى ودراسة المشيمة البشرية في الجسم الحي دون فرض مخاطر غير أخلاقية على الحمل المستمر. ولذلك، فإن دراسات وظيفة المشيمة في الإنسان تستند إلى حد كبير على نماذج في المختبر و فيفو السابقين . وقد أجريت غالبية الدراسات السابقة في الجسم الحي من نقل المشيمة والتمثيل الغذائي في الحيوانات 4 ، 5 ، 6 . ومع ذلك، كما هيكل المشيمة وظائف تختلف اختلافا كبيرا بين الأنواع، يجب أن يتم استقراء النتائج من الحيوانات للإنسان بحذر. فقط عدد قليل من الإنسان أصغر في الدراسات المجراة وقد حققت المشيمة وامتصاص الجنين والنقل في ظل الطبيعي الفسيولوجيةولم يستكشف أي منها النقل المتكامل للعديد من المركبات 7 و 8 و 9 و 10 و 11 و 12 و 13 . وتوضح هذه الدراسات الأساسية أنه في الدراسات المجراة من المشيمة البشرية هي ممكنة، وأنها قد تخدم عدة أغراض. أولا، المفاهيم الحالية لوظائف المشيمة المستمدة أساسا من في المختبر ، خارج الجسم الحي والحيوان دراسات يمكن اختبارها في بيئة الإنسان، وبالتالي توفير رؤية جديدة وأكثر تحديدا في المشيمة البشرية. ثانيا، خصائص المشيمة مختلة المرتبطة نمو الجنين الشاذة، تسمم الحمل، وسكري الأم، ومتلازمة التمثيل الغذائي وغيرها من الاضطرابات الأيضية الأمهات يمكن وصفها بشكل أفضل. ثالثا، الإنسان في الدراسات المجراة توفر فرصة لتطوير التشخيصتيك والأدوات التنبؤية من وظيفة المشيمة. على هذه الخلفية كنا تهدف إلى إنشاء مجموعة شاملة من البيانات الفسيولوجية للتحقيق وظيفة المشيمة الإنسان في الجسم الحي. خلال عملية قيصرية المخطط لها، نستغل الوصول داخل البطن إلى الوريد الرحمي لجمع عينات الدم من السفن الواردة والصادرة على الجانبين الأم والجنين من المشيمة (طريقة أخذ العينات 4-السفينة). وتستخدم هذه العينات لحساب الاختلافات تركيز الشرايين المقترنة من المواد الغذائية وغيرها من المواد 14 . بالإضافة إلى ذلك، نقيس حجم تدفق الدم على جانبي المشيمة عن طريق الموجات فوق الصوتية. وبالتالي، امتصاص المشيمة والجنين من أي مركب قد يكون كميا. وعلاوة على ذلك، فمن الممكن لتحديد المواد الصادرة عن المشيمة إلى الأمهات والجنين التعاميم 15 ، 16 ، 17 . عند الجمعد مع المعلمات السريرية للأم والطفل، وتحليلات المشيمة والأنسجة الأخرى ذات الصلة، وهذا الأسلوب لديه القدرة المثيرة لدمج العديد من جوانب وظائف المشيمة في الجسم الحي في نفس أزواج الأم والجنين.

<table><tbody><tr><td>&lt;strong&gt;Maternal body composition&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Impedance scale</td><td>Tanita</td><td></td><td>or similar</td></tr><tr><td>&lt;strong&gt;Ultrasound measurements&amp;nbsp;&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Sequoia 512 ultrasound machine</td><td>Acuson</td><td></td><td>equipped with a curved transducer with colour and pulsewave Doppler (frequency bandwidth 2 - 6 MHz)</td></tr><tr><td>&lt;strong&gt;Blood samples&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Arerial cannula</td><td>BD Medical</td><td>682245</td><td>or similar</td></tr><tr><td>20 cc Eccentric Luer Tip Syringe without Needle</td><td>Termo</td><td>SS-20ES</td><td>or similar. 3 needed.</td></tr><tr><td>10 cc Eccentric Luer Tip Syringe without Needle</td><td>Termo</td><td>SS-10ES</td><td>or similar. 9 needed.</td></tr><tr><td>5 cc 6% Luer Syringe without Needle</td><td>Termo</td><td>SS-05S1</td><td>or similar. 2 needed.</td></tr><tr><td>Arterial blood gas syringe&amp;nbsp;</td><td>Radiometer Medical</td><td></td><td>or similar. 4 needed.</td></tr><tr><td>Sterile winged needle connected to flexible tubing, 21 gauge</td><td>Greiner Bio-One</td><td>450081</td><td>(intended for single use).3 needed.</td></tr><tr><td>Vacutainer tube 6 mL EDTA&amp;nbsp;</td><td>Greiner Bio-One</td><td>456043</td><td>or similar. Label with sample site. 10 needed.</td></tr><tr><td>Vacutainer tube 5 mL LH Lithium Heparin Separator</td><td>Greiner Bio-One</td><td>456305</td><td>or similar. Label with sample site. 5 needed.</td></tr><tr><td>Vacutainer tube 6 mL Serum Clot Activator&amp;nbsp;</td><td>Greiner Bio-One</td><td>456089</td><td>or similar. Label with sample site. 5 needed.</td></tr><tr><td>Vacutainer tube 3 mL&amp;nbsp; 9NC Coagulation sodium citrate 3.2%</td><td>Greiner Bio-One</td><td>454334</td><td>or similar. Label with sample site. 5 needed.</td></tr><tr><td>Cryogenic vials, 2.0 mL</td><td>Corning</td><td>430488</td><td>or similar. Label with sample site, serum/type of plasma and ID. 90 needed.</td></tr><tr><td>Marked trays to transport the syringes</td><td></td><td></td><td>to transport the blood samples in the operation theatre</td></tr><tr><td>Rocker</td><td></td><td></td><td>for gentle mixing of the samples</td></tr><tr><td>Ice</td><td></td><td></td><td>in styrofoam box</td></tr><tr><td>Liquid nitrogen</td><td></td><td></td><td>in appropriate container</td></tr><tr><td>&lt;strong&gt;Placenta samples&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Metal tray</td><td></td><td></td><td></td></tr><tr><td>Ice</td><td></td><td></td><td>in styrofoam box</td></tr><tr><td>Calibrated scale</td><td></td><td></td><td></td></tr><tr><td>Metal ruler</td><td></td><td></td><td></td></tr><tr><td>1 M Phosphate buffered saline</td><td>Sigma</td><td>D1408</td><td>or similar. Dilute 10 M to&amp;nbsp; 1 M before use</td></tr><tr><td>RNA stabilization solution</td><td>Sigma</td><td>R0901-500ML&amp;nbsp;</td><td>or similar</td></tr><tr><td>Optimal Cutting Temperature (O.C.T.) compound</td><td>vwr</td><td>361603E</td><td>or similar</td></tr><tr><td>Cryogenic vials, 2.0 mL</td><td>Corning</td><td>430488</td><td>or similar. Label with sample site. content and ID. 10 needed.</td></tr><tr><td>Centrifuge tubes, conical bottom 50 mL</td><td>Greiner Bio-One</td><td>227,285</td><td>or similar. Label with &quot;RNA later&quot;, sample site and ID. 2 needed.</td></tr><tr><td>Liquid nitrogen</td><td></td><td></td><td>in appropriate container</td></tr><tr><td>&lt;strong&gt;Fetal body composition&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Calibrated scale</td><td></td><td></td><td></td></tr><tr><td>Measuring tape</td><td></td><td></td><td></td></tr></tbody></table>

the 4-vessel sampling approach to integrative studies of human placental physiology in vivo

المشيمة البشرية غير قابلة للوصول للغاية للبحث في حين لا يزال في الرحم . وبالتالي فإن الفهم الحالي لعلم وظائف الأعضاء المشيمة الإنسان في الجسم الحي يعتمد إلى حد كبير على الدراسات الحيوانية، على الرغم من التنوع الكبير بين الأنواع في التشريح المشيمة، ديناميكا الدم ومدة الحمل. الغالبية العظمى من الدراسات المشيمة البشرية هي دراسات نضح خارج الجسم الحي أو في المختبر دراسات الأرومة الغاذية. على الرغم من الدراسات في المختبر والنماذج الحيوانية ضرورية، استقراء النتائج من هذه الدراسات إلى المشيمة البشرية في الجسم الحي غير مؤكد. ونحن نهدف إلى دراسة علم وظائف الأعضاء المشيمة البشرية في الجسم الحي على المدى، وتقديم بروتوكول مفصل من هذه الطريقة. استغلال الدخول داخل البطن إلى الوريد الرحمي فقط قبل شق الرحم خلال العملية القيصرية المخطط لها، نقوم بجمع عينات الدم من السفن الواردة والصادرة على الجانبين الأم والجنين من المشيمة. عند الجمع بين كونقياسات الترشيح من عينات الدم مع قياسات تدفق الدم حجم، ونحن قادرون على تحديد المشيمة وامتصاص الجنين والافراج عن أي مركب. وعلاوة على ذلك، عينات الأنسجة المشيمة من نفس أزواج الأم والجنين يمكن أن توفر قياسات كثافة الناقل والنشاط والجوانب الأخرى من وظائف المشيمة في الجسم الحي . من خلال هذا الاستخدام التكاملي لطريقة أخذ العينات من 4 سفن ونحن قادرون على اختبار بعض المفاهيم الحالية لنقل المغذيات المشيمة والتمثيل الغذائي في الجسم الحي ، سواء في حالات الحمل العادية والمرضية. وعلاوة على ذلك، فإن هذه الطريقة تمكن من تحديد المواد التي يفرزها المشيمة لدورة الأمهات، والتي يمكن أن تكون مساهمة هامة في البحث عن المؤشرات الحيوية للخلل المشيمة.

طريقة أخذ العينات من 4 سفن قابلة للتطبيق في الممارسة السريرية، وحصلنا بنجاح على عينات دم من 209 أم / رضيع. في 128 من هذه حققنا أيضا لقياس حجم تدفق الدم. تم الحصول على عينات كاملة 4-السفينة وقياسات تدفق نوعية جيدة من كل من الأم والجنين الأوعية في 70 أزواج الأ...

Watch this Scientific Journal Video about The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo at JoVE.com

The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

نقدم طريقة مفصلة لدراسة علم وظائف الأعضاء المشيمة الإنسان في الجسم الحي على المدى. الأسلوب يجمع بين أخذ عينات الدم من السفن الواردة والصادرة على الجانبين الأم والجنين من المشيمة مع قياسات الموجات فوق الصوتية من حجم تدفق الدم والأوعية الأنسجة المشيمة.

نهج أخذ العينات 4-السفينة إلى الدراسات التكاملية من فسيولوجيا المشيمة الإنسان<em&gt; في فيفو</em

The human placenta is highly inaccessible for research while still in utero. The current understanding of human placental physiology in vivo is therefore ...

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Medicine

The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

10.5K Views.  Oslo University Hospital.  نقدم طريقة مفصلة لدراسة علم وظائف الأعضاء المشيمة الإنسان في الجسم الحي على المدى. الأسلوب يجمع بين أخذ عينات الدم من السفن الواردة والصادرة على الجانبين الأم والجنين من المشيمة مع قياسات الموجات فوق الصوتية من حجم تدفق الدم والأوعية...

Video: The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC twin pregnancies develop complications arising from the shared placenta and associated vascular connections1. Severe twin-to-twin syndrome (TTTS) is reported to account for approximately 20% of these complications2,3. Inter-twin vascular connections occur in almost all MC placentas and are related to the prognosis and outcome of these high-risk twin pregnancies. The number, size and type of connections have been implicated in the development of TTTS and other MC twin conditions. Three types of inter-twin vascular connections occur: 1) artery to vein connections (AVs) in which a branch artery carrying deoxygenated blood from one twin courses along the fetal surface of the placenta and dives into a placental cotyledon. Blood flows via a deep intraparenchymal capillary network into a draining vein that emerges at the fetal surface of the placenta and brings oxygenated blood toward the other twin. There is unidirectional flow from the twin supplying the afferent artery toward the twin receiving the efferent vein; 2) artery to artery connections (AAs) in which a branch artery from each twin meets directly on the superficial placental surface resulting in a vessel with pulsatile bidirectional flow, and 3) vein to vein connections (VVs) in which a branch vein from each twin meets directly on the superficial placental surface allowing low pressure bidirectional flow. In utero obstetric sonography with targeted Doppler interrogation has been used to identify the presence of AV and AA connections4. Prenatally detected AAs that have been confirmed by postnatal placental injection studies have been shown to be associated with an improved prognosis for both twins5. Furthermore, fetoscopic laser ablation of inter-twin vascular connections on the fetal surface of the shared placenta is now the preferred treatment for early, severe TTTS. 
Postnatal placental injection studies provide a valuable method to confirm the accuracy of prenatal Doppler ultrasound findings and the efficacy of fetal laser therapy6. Using colored dyes separately hand-injected into the arterial and venous circulations of each twin, the technique highlights and delineates AVs, AAs, and VVs. This definitive demonstration of MC placental vascular anatomy may then be correlated with Doppler ultrasound findings and neonatal outcome to enhance our understanding of the pathophysiology of MC twinning and its sequelae. Here we demonstrate our placental injection technique.

Vascular mapping of monochorionic (MC) twin placentas after birth provides a means for detailed demonstration of vascular connections between the twins&#x2019; circulations. 
Imbalance of these connections is thought to play a pivotal role in the development of complications of MC twinning including twin-to-twin transfusion syndrome.

دليل قسطرة أسلاك بمساعدة وحقن صبغة الملونة لرسم خرائط الأوعية الدموية للمشيمة التوأم المشيماء

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC ...

Isolation and Analysis of Hematopoietic Stem Cells from the Placenta

Hematopoietic stem cells (HSCs) have the ability to self-renew and generate all cell types of the blood lineages throughout the lifetime of an individual. All HSCs emerge during embryonic development, after which their pool size is maintained by self-renewing cell divisions. Identifying the anatomical origin of HSCs and the critical developmental events regulating the process of HSC development has been complicated as many anatomical sites participate during fetal hematopoiesis. Recently, we identified the placenta as a major hematopoietic organ where HSCs are generated and expanded in unique microenvironmental niches (Gekas, et al 2005, Rhodes, et al 2008). Consequently, the placenta is an important source of HSCs during their emergence and initial expansion.
In this article, we show dissection techniques for the isolation of murine placenta from E10.5 and E12.5 embryos, corresponding to the developmental stages of initiation of HSCs and the peak in the size of the HSC pool in the placenta, respectively. In addition, we present an optimized protocol for enzymatic and mechanical dissociation of placental tissue into single-cell suspension for use in flow cytometry or functional assays. We have found that use of collagenase for single-cell suspension of placenta gives sufficient yields of HSCs. An important factor affecting HSC yield from the placenta is the degree of mechanical dissociation prior to, and duration of, enzymatic treatment.
We also provide a protocol for the preparation of fixed-frozen placental tissue sections for the visualization of developing HSCs by immunohistochemistry in their precise cellular niches. As hematopoietic specific antigens are not preserved during preparation of paraffin embedded sections, we routinely use fixed frozen sections for localizing placental HSCs and progenitors.

We have identified the placenta as a major hematopoietic organ during development. We found that hematopoietic stem cells (HSCs) are both generated and expanded in the placenta in unique microenvironmental niches. Here, we describe experimental techniques required for isolation and visualization of HSCs in the mouse placenta.

العزلة وتحليل الخلايا الجذعية المكونة للدم من المشيمة

Hematopoietic stem cells (HSCs) have the ability to self-renew and generate all cell types of the blood lineages throughout the lifetime of an individual. ...

Biology

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model

Decades ago the human placenta was thought to be an impenetrable barrier between mother and unborn child. However, the discovery of thalidomide-induced birth defects and many later studies afterwards proved the opposite. Today several harmful xenobiotics like nicotine, heroin, methadone or drugs as well as environmental pollutants were described to overcome this barrier. With the growing use of nanotechnology, the placenta is likely to come into contact with novel nanoparticles either accidentally through exposure or intentionally in the case of potential nanomedical applications. Data from animal experiments cannot be extrapolated to humans because the placenta is the most species-specific mammalian organ 1. Therefore, the ex vivo dual recirculating human placental perfusion, developed by Panigel et al. in 1967 2 and continuously modified by Schneider et al. in 1972 3, can serve as an excellent model to study the transfer of xenobiotics or particles.
Here, we focus on the ex vivo dual recirculating human placental perfusion protocol and its further development to acquire reproducible results.
The placentae were obtained after informed consent of the mothers from uncomplicated term pregnancies undergoing caesarean delivery. The fetal and maternal vessels of an intact cotyledon were cannulated and perfused at least for five hours. As a model particle fluorescently labelled polystyrene particles with sizes of 80 and 500 nm in diameter were added to the maternal circuit. The 80 nm particles were able to cross the placental barrier and provide a perfect example for a substance which is transferred across the placenta to the fetus while the 500 nm particles were retained in the placental tissue or maternal circuit. The ex vivo human placental perfusion model is one of few models providing reliable information about the transport behavior of xenobiotics at an important tissue barrier which delivers predictive and clinical relevant data.

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the ex vivo Human Placental Perfusion Model

The ex vivo dual recirculating human placental perfusion model can be used to investigate the transfer of xenobiotics and nanoparticles across the human placenta. In this video protocol we describe the equipment and techniques required for a successful execution of a placenta perfusion.

تحديد معدل النقل من الاكسيوبيوتك والمواد النانوية عبر المشيمة باستخدام<em&gt; خارج الحي</em&gt; الإنسان المشيمة الإرواء الموديل

Decades ago the human placenta was thought to be an impenetrable barrier between mother and unborn child. However, the discovery of thalidomide-induced ...

Bioengineering

Isolation of Cellular Lipid Droplets: Two Purification Techniques Starting from Yeast Cells and Human Placentas

Lipid droplets are dynamic organelles that can be found in most eukaryotic and certain prokaryotic cells. Structurally, the droplets consist of a core of neutral lipids surrounded by a phospholipid monolayer. One of the most useful techniques in determining the cellular roles of droplets has been proteomic identification of bound proteins, which can be isolated along with the droplets. Here, two methods are described to isolate lipid droplets and their bound proteins from two wide-ranging eukaryotes: fission yeast and human placental villous cells. Although both techniques have differences, the main method - density gradient centrifugation -&#160;is shared by both preparations. This shows the wide applicability of the presented droplet isolation techniques.

In the first protocol, yeast cells are converted into spheroplasts by enzymatic digestion of their cell walls. The resulting spheroplasts are then gently lysed in a loose-fitting homogenizer. Ficoll is added to the lysate to provide a density gradient, and the mixture is centrifuged three times. After the first spin, the lipid droplets are localized to the white-colored floating layer of the centrifuge tubes along with the endoplasmic reticulum (ER), the plasma membrane, and vacuoles. Two subsequent spins are used to remove these other three organelles. The result is a layer that has only droplets and bound proteins.

In the second protocol, placental villous cells are isolated from human term placentas by enzymatic digestion with trypsin and DNase I. The cells are homogenized in a loose-fitting homogenizer. Low-speed and medium-speed centrifugation steps are used to remove unbroken cells, cellular debris, nuclei, and mitochondria. Sucrose is added to the homogenate to provide a density gradient and the mixture is centrifuged to separate the lipid droplets from the other cellular fractions.

The purity of the lipid droplets in both protocols is confirmed by Western Blot analysis. The droplet fractions from both preps are suitable for subsequent proteomic and lipidomic analysis.

Two techniques for isolating cellular lipid droplets from 1) yeast cells and 2) human placentas are presented. The centerpiece of both procedures is density gradient centrifugation, where the resulting floating layer containing the droplets can be readily visualized by eye, extracted, and quantified by Western Blot analysis for purity.

عزل الخلوية الدهون قطرات: اثنان تقنيات تنقية بدءا من خلايا الخميرة ومشيمة الآدمي

Lipid droplets are dynamic organelles that can be found in most eukaryotic and certain prokaryotic cells. Structurally, the droplets consist of a core of ...

Three-dimensional Rendering and Analysis of Immunolabeled, Clarified Human Placental Villous Vascular Networks

Nutrient and gas exchange between mother and fetus occurs at the interface of the maternal intervillous blood and the vast villous capillary network that makes up much of the parenchyma of the human placenta. The distal villous capillary network is the terminus of the fetal blood supply after several generations of branching of vessels extending out from the umbilical cord. This network has a contiguous cellular sheath, the syncytial trophoblast barrier layer, which prevents mixing of fetal blood and the maternal blood in which it is continuously bathed. Insults to the integrity of the placental capillary network, occurring in disorders such as maternal diabetes, hypertension and obesity, have consequences that present serious health risks for the fetus, infant, and adult. To better define the structural effects of these insults, a protocol was developed for this study that captures capillary network structure on the order of 1 - 2 mm3 wherein one can investigate its topological features in its full complexity. To accomplish this, clusters of terminal villi from placenta are dissected, and the trophoblast layer and the capillary endothelia are immunolabeled. These samples are then clarified with a new tissue clearing process which makes it possible to acquire confocal image stacks to z- depths of ~1 mm. The three-dimensional renderings of these stacks are then processed and analyzed to generate basic capillary network measures such as volume, number of capillary branches, and capillary branch end points, as validation of the suitability of this approach for capillary network characterization.

This study presents a protocol for the reversible tissue clearing, immunostaining, 3D-rendering and analysis of vascular networks in human placenta villi samples on the order of 1 - 2 mm3.

التقديم ثلاثي الأبعاد وتحليل إيمونولابيليد، أوضح البشري شبكات الأوعية الدموية فيلوس المشيمية

Nutrient and gas exchange between mother and fetus occurs at the interface of the maternal intervillous blood and the vast villous capillary network that ...

Developmental Biology

Ex Vivo Perfusion of the Rodent Placenta

The placenta is a key organ during pregnancy that serves as a barrier to fetal xenobiotic exposure and mediates the exchange of nutrients for waste. An assay is described here to perfuse an isolated rat placenta and evaluate the maternal-to-fetal translocation of xenobiotics ex vivo. In addition, the evaluation of physiological processes such as fluid flow to the fetus and placental metabolism may be conducted with this methodology. This technique is suitable for evaluating maternal-to-fetal kinetics of pharmaceutical candidates or environmental contaminants. In contrast to current alternative approaches, this methodology allows the evaluation of the isolated maternal-fetal vasculature, with the systemic neural or immune involvement removed, allowing any observed changes in physiological function to be attributable to local factors within the isolated tissue.

Here is presented a protocol of ex vivo maternal-fetal vascular perfusion to enable the administration of a test article into maternal vasculature and to evaluate placental transfer of xenobiotic particles or pharmacological agents in addition to alterations in placental physiology.

السابقين فيفو Perfusion من المشيمة القوارض

The placenta is a key organ during pregnancy that serves as a barrier to fetal xenobiotic exposure and mediates the exchange of nutrients for waste. An ...

Human Placental and Decidual Organ Cultures to Study Infections at the Maternal-fetal Interface

The placenta shows a large degree of interspecies anatomic variability. To best understand biology and pathophysiology of the human placenta, it is imperative to design experiments using human cells and tissues. An advantage of organ culture is maintenance of three-dimensional (3D) structural organization and extracellular matrix. The goal of the method described here is successful establishment of ex vivo human gestational tissue organ cultures and their healthy culture maintenance for 72-96 hr. The protocol details the immediate processing of research-consented, placental and decidual specimens fresh from the operating suite. These are abundant specimens that would otherwise be discarded. Detailed instructions on the sterile collection of these samples, including morphologic details on how to select appropriate tissues to establish 3D organ cultures, is provided. Placental villous and decidual tissues are microdissected into 2-3 mm3 pieces and placed separately on matrix-lined transwell filters and cultured for several days. Villous and decidual organ cultures are well suited for the study of human host-pathogen interaction. As compared to other model organisms, these human cultures are particularly advantageous to examine mechanism of infection for pathogens that demonstrate variable patterns of host specificity. As an example, we demonstrate infection of placental and decidual organ cultures with the clinically relevant, facultative intracellular bacterial pathogen Listeria monocytogenes.

A simple method to establish primary human placental (villous) and decidual organ cultures is described. Villous and decidual organ cultures are invaluable tools for studying pathogenesis at the human maternal-fetal interface. Infection with the facultative intracellular bacterium Listeria monocytogenes is demonstrated.

الثقافات المشيمة وساقطي الجهاز الإنسان لدراسة العدوى في واجهة الأم والجنين

The placenta shows a large degree of interspecies anatomic variability. To best understand biology and pathophysiology of the human placenta, it is ...

Immunology and Infection

تحسين زراعة المشيمة خارج الجسم الحي باستخدام نهج قائم على التدفق

Ex Vivo Placental Explant Flow Culture - Mimicking the Dynamic Conditions In Utero

The existing ex vivo placental explant culture models are primarily grounded in static culture systems using well plates. However, these models inadequately reflect the dynamic in utero setting, where the placenta encounters constant slight shear stress due to plasma or blood flow. To address this limitation, a flow culture system has been devised to bring ex vivo placental explant cultivation closer to the in utero flow conditions experienced within the maternal body. Within this approach, placental explants are cultivated in a sequence of five interconnected flow chambers. This setting&#160;maintains physiological oxygen concentrations and a consistent flow rate. The collected data reveals that under flow conditions, the preservation of tissue morphology exhibits notable enhancement compared to conventional static methods. This innovative technique introduces a straightforward means of ex vivo placental explant culture, offering a more faithful representation of the dynamic in vivo environment. Moreover, this study introduces new&#160;possibilities for investigating the functional dynamics of the feto-maternal interface. By embracing feasible dynamic methodologies, a deeper comprehension of placental biology is facilitated, underscoring its relevance for maternal-fetal health.

تسليط الضوء على المؤلف: تطوير البحث من خلال تسلسل الخلية المفردة والأنسجة المكانية في أنسجة المشيمة 

Here is a protocol for culturing placental explants under constant flow conditions. This approach enhances traditional static villous culture systems by enabling the replication of dynamic physiological environments.

نهج أخذ العينات 4-السفينة إلى الدراسات التكاملية من فسيولوجيا المشيمة الإنسان

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