Name: application of hemostatic devices in laparoscopic hepatectomy
Uploaded: 2022-04-19T13:00:00
Description: Watch this Scientific Journal Video about Application of Hemostatic Devices in Laparoscopic Hepatectomy at JoVE.com

Bu çalışma, Guangzhou Şehri Bilim ve Teknoloji Projesi'nden (202102010090) gelen hibelerle desteklenmiştir.

Operasyon rutindir ve etik onay almıştır. Onaylar aşağıdaki gibidir: Guangzhou Tıp Üniversitesi İkinci Bağlı Hastanesi Klinik Araştırma ve Uygulama Etiği Komitesi tarafından hızlı inceleme onayı: Bilimsel araştırma etiği inceleme projesi "Laparoskopik Hepatektomide Basit ve Kolay Hemostatik Cihazların Uygulanması" (kabul No: 2022-hg-ks-02), Guangzhou Tıp Üniversitesi İkinci Bağlı Hastanesi'nin klinik araştırma ve uygulama etiği komitesi tarafından onaylanmıştır. Araştırma içeriği ve...

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Y., Han, H. S., Yoon, Y. S., Shin, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feasibility+of+laparoscopic+liver+resection+for+tumors+located+in+the+posterosuperior+segments+of+the+liver,+with+a+special+reference+to+overcoming+current+limitations+on+tumor+location.">Feasibility of laparoscopic liver resection for tumors located in the posterosuperior segments of the liver, with a special reference to overcoming current limitations on tumor location.</a> Surgery. 144 (1), 32-38 (2008).</li><li>Ome, Y., Honda, G., Doi, M., Muto, J., Seyama, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+anatomic+liver+resection+of+segment+8+using+intrahepatic+Glissonean+approach.">Laparoscopic anatomic liver resection of segment 8 using intrahepatic Glissonean approach.</a> Journal of the American College of Surgeons. 230 (3), 13-20 (2020).</li><li>Ryu, T., 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=Perioperative+and+oncological+outcomes+of+laparoscopic+anatomical+hepatectomy+for+hepatocellular+carcinoma+introduced+gradually+in+a+single+center.">Perioperative and oncological outcomes of laparoscopic anatomical hepatectomy for hepatocellular carcinoma introduced gradually in a single center.</a> Surgical Endoscopy. 32 (2), 790-798 (2018).</li><li>Makabe, K., 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=Efficacy+of+occlusion+of+hepatic+artery+and+risk+of+carbon+dioxide+gas+embolism+during+laparoscopic+hepatectomy+in+a+pig+model.">Efficacy of occlusion of hepatic artery and risk of carbon dioxide gas embolism during laparoscopic hepatectomy in a pig model.</a> Journal of Hepato-Biliary-Pancreatic Sciences. 21 (8), 592-598 (2014).</li><li>Miyagi, S., 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=Pure+laparoscopic+hepatectomy+combined+with+a+pure+laparoscopic+pringle+maneuver+in+patients+with+severe+cirrhosis.">Pure laparoscopic hepatectomy combined with a pure laparoscopic pringle maneuver in patients with severe cirrhosis.</a> Case Reports in Gastroenterology. 9 (1), 101-105 (2015).</li></ol>

Hepatektomi, primer karaciğer kanseri ve sekonder karaciğer kanseri olan hastalar için en önemli tedavi yöntemlerinden biridir. 1991 yılında Reich ve ark. karaciğerin iyi huylu tümörlerini çıkarmak için laparoskopi uyguladılar ve dünyanın ilk laparoskopik hepatektomisini tamamladılar4. Yıllar süren hızlı gelişim ve tanıtımdan sonra, laparoskopik hepatektomi karaciğer malign tümörlerinin tedavisinde ana akım yöntemlerden biri haline gelmiş ve son yıllarda yaygın olar...

Hepatosellüler karsinom, sindirim sisteminin en sık görülen malign tümörlerinden biridir. Farklı primer bölgelere göre, primer hepatosellüler karsinom ve sekonder karaciğer kanseri olarak ayrılabilir. Hücreler, karaciğer dışındaki organlardan karaciğere giden primer tümörlerden karaciğere çeşitli şekillerde metastaz yapabilir ve karaciğerde karsinoma yol açabilir. Karaciğerdeki metastatik hücrelerin% 50'sinden fazlasının kolorektal kanserden geldiği, diğerlerinin ise meme, pankreas, akciğer ve mide tümörlerinden geldiği bildirilmiştir1. Son yıllarda, sistematik kemoterapi, girişimsel tedavi, moleküler hedefli tedavi, cerrahi vb. dahil olmak üzere sekonder karaciğer kanseri için birçok tedavi mevcuttur2. Bununla birlikte, radikal rezeksiyon hala en etkili tedavidir çünkü kanseri tamamen ortadan kaldırabilir3.
Laparoskopik teknolojinin hızla gelişmesiyle birlikte, laparoskopik anatomik hepatektomi cerrahlar tarafından yavaş yavaş tanınmakta, ancak özellikle birincil tıp kurumlarında hala yaygın olarak uygulanmamaktadır. Bunun nedenlerinden biri, yüksek dereceli hemostatik cihazlara duyulan gereksinimdir. Bunlar operasyon sürecinde kanama ve safra kaçağı risklerini azaltmak için gereklidir. Burada, laparoskopik hepatektomi için tek lümen kateter, harmonik neşter ve monopol elektrokoagülasyon dahil olmak üzere bir dizi basit ve kolay hemostatik ekipman sunulmaktadır. Bunu yapmak için, önce porta hepatis tek bir lümen kateter kullanılarak aralıklı olarak tıkanır. Karaciğer parankim dokusu daha sonra harmonik neşter kullanılarak rezeke edilir. Kanama lekeleri monopol elektrokoagülasyon ile noktadan noktaya pıhtılaşır. Bu basit ve kolay hemostatik ekipman, pringle manevrasını gerçekleştirmek için tek bir lümen kateter kullanır ve harmonik neşter hemostazı ve monopol elektrokoagülasyonunu kullanır. Ekipman hastanelerde kolayca bulunabilir, böylece laparoskopi veya eğitim için ek kolaylık sağlar. Bu nedenle, bu basit ve kolay hemostatik cihazlar, birincil tıbbi kurumlarda prosedürlerin yürütülmesi için uygundur.
Bu çalışmada hasta, karaciğerde metastaz yapmış sigmoid kolonun orta derecede diferansiye adenokarsinomu tanısı alan 67 yaşında bir erkekti. Sigmoid kolon kanserinin radikal rezeksiyonu Ocak 2021'de yapıldı. Patolojik sonuçlar pT4aN2aM1'in TNM evresi ile orta derecede diferansiye adenokarsinom idi. Ameliyattan sonra dört kez FOLFOX kemoterapisi verildi. Bunu takiben, karaciğer metastazlarının lezyonunu tamamen ortadan kaldırmak için hastanın vücut durumu laparoskopik anatomik hepatektomi yapmaya uygun görüldü. Karaciğer fonksiyonunun Child-Pugh derecesi Grade A idi. Karaciğer rezerv fonksiyon testi için, ICG klirens testinde R15% 1.6 (<% 10) idi. BT, S5'in dorsal kısmında ve karaciğerin S6'sının ventral kısmında 57 mm x 68 mm x 76 mm'lik bir tümör gösterdi; üç boyutlu rekonstrüksiyon modeli Şekil 1'de gösterilmiştir.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>HARMONIC ACE Ultrasonic Surgical Devices</td>
			<td>Ethicon Endo-Surgery,LLC</td>
			<td>HAR36</td>
			<td></td>
		</tr>
		<tr>
			<td>Hem-o-lock</td>
			<td>Teleflex Medical</td>
			<td>544233</td>
			<td></td>
		</tr>
		<tr>
			<td>Monopole Electrocoagulation</td>
			<td>KANGJI MEDICAL</td>
			<td>/</td>
			<td></td>
		</tr>
		<tr>
			<td>single lumen ureter</td>
			<td>WELL LEAD MEDICAL CO.,LTD.</td>
			<td>12F</td>
			<td></td>
		</tr>
	</tbody>
</table>

application of hemostatic devices in laparoscopic hepatectomy

Laparoskopik hepatektomi, minimal invaziv bir yöntem olması nedeniyle benign ve malign karaciğer hastalıklarının tedavisinde geleneksel bir yöntem olarak kabul edilir. Non-invaziv yönüne rağmen, ameliyat sırasında veya ameliyat sonrası dönemde karaciğer parankimi doku rezeksiyonunda kanama ve safra kaçağı meydana gelir, bu da ultrasonik cerrahi aspirasyon, bipolar elektrokoagülasyon gibi yüksek dereceli hemostatik cihazlara ihtiyaç duyulduğunu gösterir. Bu yüksek dereceli hemostatik cihazların bulunmaması, laparoskopik hepatektominin temel tıbbi kuruluşlarda genelleştirilmiş bir prosedür haline gelmesini engellemektedir. Yukarıda belirtilen durum göz önüne alındığında, karaciğer parankim dokusu rezeksiyonunu yenilikçi bir şekilde gerçekleştirmek için harmonik bir neşter, monopol elektrokoagülasyon ve tek bir lümen kateter içeren bu protokolde basit ve kolay hemostatik cihazlar paketi geliştirilmiştir. Her şeyden önce, porta hepatis veya hepatik pedikül, tek bir lümen kateter tarafından aralıklı olarak tıkanır, ardından 15 dakika boyunca kelepçelenir ve 5 dakika boyunca serbest bırakılır. Daha sonra, harmonik neşter kullanılarak, hepatik parankim dokusunu kesmek ve intrahepatik arterleri, damarları ve safra kanallarını ortaya çıkarmak için karaciğerin kelepçelenmesi ve ezilmesi yapılır. Son olarak, kanama lekeleri her noktada monopol elektrokoagülasyon kullanılarak pıhtılaştırılır. İntrahepatik boru hattı yapıları daha sonra kanamayı kolayca durdurabilen, safra kaçağı insidansını azaltabilen ve laparoskopik hepatektominin güvenliğini ve fizibilitesini artırabilen bu yöntemler kullanılarak görülebilir. Bu nedenle, burada gösterilen basit ve kolay hemostatik cihazlar, birincil tıbbi kurumlarda prosedürlerin yürütülmesi için uygundur.

Ameliyat 2.5 saat içinde tamamlandı ve intraoperatif kanama hacmi kan transfüzyonu yapılmadan 100 mL idi. Kısa süreli komplikasyon görülmedi ve hasta ameliyattan sonraki 8. günde hastaneden taburcu edildi. Ameliyat sonrası karsinoembriyonik antijen (CEA) düzeyi 1058.69 μg/L olan ameliyat öncesi seviyeden 110.64 μg/L'ye düşmüştür (Şekil 12). Ameliyattan iki ay sonra, hasta kemoterapi tedavisine devam etmek için hastaneye döndü (kemoterapi rejimi mFOLFOX6 idi)...

Watch this Scientific Journal Video about Application of Hemostatic Devices in Laparoscopic Hepatectomy at JoVE.com

Application of Hemostatic Devices in Laparoscopic Hepatectomy

Laparoskopik hepatektomi için yüksek dereceli hemostatik cihazlar gereklidir. Bununla birlikte, bu cihazlar temel tıbbi organizasyonlarda genelleştirilmemiştir. Bu nedenle, bu makalede, laparoskopik hepatektominin gerçekleştirilmesini kolaylaştırabilecek basit ve kolay hemostatik cihazların bir paketi gösterilmiştir.

Laparoskopik Hepatektomide Hemostatik Cihazların Uygulanması

Laparoscopic hepatectomy is considered a conventional method for treating benign and malignant liver diseases because it is a minimally invasive method. ...

Bu tekniğin temel avantajı, hemostatik cihazın basit ve elde edilmesi kolay olması ve temel hastanelerde gerçekleştirilebilmesidir. Bu hemostatik ekipman seti ayrıca laparoskopik hepatektomi ve diğer sistemlere uygulanabilir. Laparoskopik cerrahinin belirli bir temeline sahip olmak, özellikle de laparoskopik hepatektomide bu hemostatik cihaz setini uygulamak için harmonik bir neşterin yetenekli kullanımı şiddetle tavsiye edilir.Laparoskopide pnömoperitoneum oluşturmak için doğrudan delinme uygulayarak başlayın. Subumbilikal bölgedeki her gözlem deliğine, kostal marjın ve ventral orta hattın altındaki sağ orta klaviküler çizgiye 10 milimetrelik bir trokar yerleştirin. Daha sonra sağ ön yardımcı hatta ve ventral orta hatta beş milimetrelik bir trokar yerleştirin.Harmonik neşter, monopol elektrokoagülasyon ve tek lümen kateterinden başlayarak hemostatik cihazları karaciğer parankim doku rezeksiyonunda yenilikçi bir şekilde kullanılmak üzere monte edin. Bir Pringle manevrası yapmak için, tek bir lümen kateter kullanarak ilk porta hepatisi aralıklı olarak tıkayın. Lümen kateteri hemostatik bir klipsle 15 dakika boyunca kelepçeleyin, ardından klipsi kesmek için harmonik neşter kullanarak beş dakika boyunca serbest bırakın ve ilk porta hepatis'e kan akışını sağlamak için klipsi çıkarın.Tümörü besleyen hepatik pedikülleri ayırın. Bunları harmonik bir neşterle tek tek parçalayın ve bölün. Hepatik pediküllerin rezeksiyonundan sonra görünür hale gelen karaciğer yüzeyindeki iskemi çizgisini gözlemleyin.İşaretlemek için kancayı kullanın. Karaciğeri kademeli olarak sıkıştırıp ezerken harmonik neşter kullanarak karaciğer parankim dokusunu rezeke edin. İç hepatik arteri, damarı ve safra kanalını gözlemleyin.Bu boru hattı yapılarını parçalayın ve bölün. Kanama ortaya çıktığında, noktadan noktaya monopol elektrokoagülasyonunu kullanarak karaciğer diseksiyon yüzeyindeki kanama lekelerini derhal pıhtılaştırın. Sağ posterior hepatik pedikülü intrahepatik parankim doku rezeksiyonu için bir işaret olarak kullanın.Ana posterior hepatik pedikülü ayırın ve tümörü besleyen pediküllerin dalını diseke edin. Operasyonu tamamlamak için, karaciğer yüzeyini yıkayın, böylece hala kanama lekeleri olup olmadığını görebilirsiniz. Karaciğer diseksiyon yüzeyindeki kanamayı durdurun ve tümör örneğini karın boşluğundan çıkarın.Operasyon 2,5 saat içinde tamamlandı. İntraoperatif kanama hacmi kan transfüzyonu yapılmadan 100 mililitre idi ve kısa süreli komplikasyon gözlenmedi. Ameliyat sonrası karsinoembriyonik antijen seviyesi, ameliyat öncesi litre başına 1058.69 mikrogram seviyesinden litre başına 110.64 mikrograma düşmüştür.Tümör patolojisi sonuçlarına göre ameliyattan iki ay sonra kemoterapi tedavisine devam edildi. Postoperatif BT incelemesi preoperatif BT ile karşılaştırıldı ve tümör büyümesi bulgusu gözlenmedi. Cerrahi süreçte en önemli adım harmonik neşter kullanılmasıdır.Hayati yapılara zarar vermemek için iyi miktarda beceri gereklidir.

application-of-hemostatic-devices-in-laparoscopic-hepatectomy

Research

JoVE Journal

Cancer Research

Evaporation-reducing Culture Condition Increases the Reproducibility of Multicellular Spheroid Formation in Microtiter Plates

Tumor models that closely imitate in vivo conditions are becoming increasingly popular in drug discovery and development for the screening of potential anti-cancer drugs. Multicellular tumor spheroids (MCTSes) effectively mimic the physiological conditions of solid tumors, making them excellent in vitro models for lead optimization and target validation. Out of the various techniques available for MCTS culture, the liquid-overlay method on agarose is one of the most inexpensive methods for MCTS generation. However, the reliable transfer of MCTS cultures using liquid-overlay for high-throughput screening may be compromised by a number of limitations, including the coating of microtiter plates (MPs) with agarose and the irreproducibility of uniform MCTS formation across wells. MPs are significantly prone to edge effects that result from excessive evaporation of medium from the exterior of the plate, preventing the use of the entire plate for drug tests. This manuscript provides detailed technical improvements to the liquid-overlay technique to increase the scalability and reproducibility of uniform MCTS formation. Additionally, details on a simple, semi-automatic, and universally applicable software tool for the evaluation of MCTS features after drug treatment is presented.

The uneven loss of medium from microtiter plates affects the reproducibility of uniform multicellular tumor spheroid formation. Improving culture conditions to reduce significant medium loss will improve the reproducibility of spheroid formation and the results of spheroid-based assays using the liquid-overlay technique.

Buharlaşma azaltıcı Kültür Durumu Mikrotiter Tabaklar Multisellüler Sfero Oluşumunun Tekrarlanabilirlik artırır

Tumor models that closely imitate in vivo conditions are becoming increasingly popular in drug discovery and development for the screening of potential ...

Kanser Araştırmaları

Application of Laser Micro-irradiation for Examination of Single and Double Strand Break Repair in Mammalian Cells

Highly coordinated DNA repair pathways exist to detect, excise and replace damaged DNA bases, and coordinate repair of DNA strand breaks. While molecular biology techniques have clarified structure, enzymatic functions, and kinetics of repair proteins, there is still a need to understand how repair is coordinated within the nucleus. Laser micro-irradiation offers a powerful tool for inducing DNA damage and monitoring the recruitment of repair proteins. Induction of DNA damage by laser micro-irradiation can occur with a range of wavelengths, and users can reliably induce single strand breaks, base lesions and double strand breaks with a range of doses. Here, laser micro-irradiation is used to examine repair of single and double strand breaks induced by two common confocal laser wavelengths, 355 nm and 405 nm. Further, proper characterization of the applied laser dose for inducing specific damage mixtures is described, so users can reproducibly perform laser micro-irradiation data acquisition and analysis.

Confocal fluorescence microscopy and laser micro-irradiation offer tools for inducing DNA damage and monitoring the response of DNA repair proteins in selected sub-nuclear areas. This technique has significantly advanced our knowledge of damage detection, signaling, and recruitment. This manuscript demonstrates these technologies to examine single and double strand break repair.

Lazer mikro-ile kullanma için tek incelenmesi ve çift Strand Break onarım memeli hücrelerinde

Highly coordinated DNA repair pathways exist to detect, excise and replace damaged DNA bases, and coordinate repair of DNA strand breaks. While molecular ...

Laparoscopic Pancreatoduodenectomy With Modified Blumgart Pancreaticojejunostomy

Minimally invasive pancreatic resections are technically demanding but rapidly increasing in popularity. In contrast to laparoscopic distal pancreatectomy, laparoscopic pancreatoduodenectomy (LPD) has not yet obtained wide acceptance, probably due to technical challenges, especially regarding the pancreatic anastomosis.
The study describes and demonstrates all steps of LPD, including the modified Blumgart pancreaticojejunostomy. Indications for LPD are all pancreatic and peri-ampullary tumors without vascular involvement. Relative contra-indications are body mass index &#62;35 kg/m2, chronic pancreatitis, mid-cholangiocarcinomas and large duodenal cancers.
The patient is in French position, 6 trocars are placed, and dissection is performed using an (articulating) sealing device. A modified Blumgart end-to-side pancreaticojejunostomy is performed with 4 large needles (3/0) barbed trans-pancreatic sutures and 4 to 6 duct-to-mucosa sutures using 5/0 absorbable multifilament combined with a 12 cm, 6 or 8 Fr internal stent using 3D laparoscopy. Two surgical drains are placed alongside the pancreaticojejunostomy.
The described technique for LPD including a modified Blumgart pancreatico-jejunostomy is well standardized, and its merits are currently studied in the randomized controlled multicenter trial. This complex operation should be performed at high-volume centers where surgeons have extensive experience in both open pancreatic surgery and advanced laparoscopic gastro-intestinal surgery.

Laparoscopic pancreatoduodenctomy (LPD) may offer advantages over open pancreatoduodenectomy, including early postoperative mobilization, less delayed gastric emptying and a shorter hospital stay. However, LPD is technically challenging and not well-standardized, especially regarding the pancreatic anastomosis. We describe a standardized technique for the pancreatic anastomosis during LPD: modified Blumgart pancreaticojejunostomy.

Değiştirilmiş Blumgart Pancreaticojejunostomy ile laparoskopik Pancreatoduodenectomy

Minimally invasive pancreatic resections are technically demanding but rapidly increasing in popularity. In contrast to laparoscopic distal ...

The Application of 1% Methylene Blue Dye As a Single Technique in Breast Cancer Sentinel Node Biopsy

In this study, we injected 1% methylene blue dye (MBD) into the subareolar or peritumoral space of the breast. In the case of breast conserving surgery (BCS), a separate incision in the lower axilla hairline was made to find the sentinel nodes (SNs). In mastectomy, the SNs were identified through the same mastectomy incision. The SNs were described as blue nodes or nodes with lymphatic blue channels. An anatomical landmark in the axilla was used to facilitate SNs identification. The SNs metastases were evaluated by intraoperative frozen section analysis and histopathology examination as it is a gold standard. Here, we described the MBD as the lone technique in breast cancer sentinel node biopsy (SNB) which could be useful when radioisotope tracer or patent or isosulfan blue dye (PBD) cannot be provided.

In Indonesia, sentinel node biopsy (SNB) is not routinely performed for breast cancer surgery because of the limitation to provide radioisotope tracer and isosulfan or patent blue dye (PBD). To overcome this obstacle, we applied 1% methylene blue dye (MBD) as a single agent to map the sentinel nodes (SNs).

Meme kanseri Sentinel düğüm biyopsi tek bir tekniği olarak 1% metilen mavi boya uygulaması

In this study, we injected 1% methylene blue dye (MBD) into the subareolar or peritumoral space of the breast. In the case of breast conserving surgery ...

The Clinical Application of Tumor Treating Fields Therapy in Glioblastoma

Glioblastoma is the most common and lethal form of brain cancer, with a median survival of 15 months after diagnosis and a 5 year survival rate of only 5% with current standard of care. Tumors often recur within 9 months following initial surgery, radiation and chemotherapy, at which point treatment options become limited. This highlights the pressing need for the development of better therapeutics to prolong survival and increase the quality of life for these patients.
Tumor Treating Fields (TTFields) therapy was developed to take advantage of the effect of low frequency alternating electrical fields on cells for cancer therapy. TTFields have been demonstrated to disrupt cells during mitosis and slow tumor growth. There is also growing evidence that they act through stimulating immune responses within exposed tumors. The advantages of TTFields therapy include its noninvasive approach and increased quality of life compared to other treatment modalities such as cytotoxic chemotherapies. The Food and Drug Administration approved TTFields therapy for the treatment of recurrent glioblastoma in 2011 and for newly diagnosed glioblastoma in 2015. We report on the effects of TTFields during mitosis, the results of electric fields modeling, and proper transducer array placement. Our protocol outlines the clinical application of TTFields on a patient post-surgery, using the second-generation device.

Glioblastoma is the most common and aggressive primary brain malignancy in adults, with most tumors recurring after initial treatment. Tumor Treating Fields (TTFields) therapy is the newest treatment modality for glioblastoma. Here, we describe the proper application of TTFields-transducer arrays on patients and discuss theory and aspects of treatment.

Glioblastoma içinde alanları terapi tedavi tümör klinik uygulama

Glioblastoma is the most common and lethal form of brain cancer, with a median survival of 15 months after diagnosis and a 5 year survival rate of only 5% ...

In Vitro Evaluation of Oncogenic Transformation in Human Mammary Epithelial Cells

Tumorigenesis is a multi-step process in which cells acquire capabilities&#160;that allow their growth, survival, and dissemination under hostile conditions. Different tests seek to identify and quantify these hallmarks of cancerous cells; however, they often focus on a single aspect of cellular transformation and, in fact, multiple tests are required for their proper characterization. The purpose of this work is to provide researchers with a set of tools to assess cellular transformation in vitro from a broad perspective, thereby making it possible to draw sound conclusions.
A sustained proliferative signaling activation is the major feature of tumoral tissues and can be easily monitored under in vitro conditions by calculating the number of population doublings achieved over time. Besides, the growth of cells in 3D cultures allows their interaction with surrounding cells, resembling what occurs in vivo. This enables the evaluation of cellular aggregation and, together with immunofluorescent labeling of distinctive cellular markers, to obtain information on another relevant feature of tumoral transformation: the loss of proper organization. Another remarkable characteristic of transformed cells is their capacity to grow without attachment to other cells and to the extracellular matrix, which can be evaluated with the anchorage assay.
Detailed experimental procedures to evaluate cell growth rate, to perform immunofluorescent labeling of cell lineage markers in 3D cultures, and to test anchorage-independent cell growth in soft agar are provided. These methodologies are optimized for Breast Primary Epithelial Cells (BPEC) due to its relevance in breast cancer; however, procedures can be applied to other cell types after some adjustments.

This protocol provides experimental in vitro tools to evaluate the transformation of human mammary cells. Detailed steps to follow-up cell proliferation rate, anchorage-independent growth capacity, and distribution of cell lineages in 3D cultures with basement membrane matrix are described.

İnsan Meme Epitel Hücrelerinde Onkojenik Dönüşümün İn Vitro Değerlendirilmesi

Tumorigenesis is a multi-step process in which cells acquire capabilities&#160;that allow their growth, survival, and dissemination under hostile ...

Co-culture of Glutamatergic Neurons and Pediatric High-Grade Glioma Cells Into Microfluidic Devices to Assess Electrical Interactions

Pediatric high-grade gliomas (pHGG) represent childhood and adolescent brain cancers that carry a rapid dismal prognosis. Since there is a need to overcome the resistance to current treatments and find a new way of cure, modeling the disease as close as possible in an in vitro setting to test new drugs and therapeutic procedures is highly demanding. Studying their fundamental pathobiological processes, including glutamatergic neuron hyperexcitability, will be a real advance in understanding interactions between the environmental brain and pHGG cells. Therefore, to recreate neurons/pHGG cell interactions, this work shows the development of a functional in vitro model co-culturing human-induced Pluripotent Stem (hiPS)-derived cortical glutamatergic neurons pHGG cells into compartmentalized microfluidic devices and a process to record their electrophysiological modifications. The first step was to differentiate and characterize human glutamatergic neurons. Secondly, the cells were cultured in microfluidic devices with pHGG derived cell lines. Brain microenvironment and neuronal activity were then included in this model to analyze the electrical impact of pHGG cells on these micro-environmental neurons. Electrophysiological recordings are coupled using multielectrode arrays (MEA) to these microfluidic devices to mimic physiological conditions and to record the electrical activity of the entire neural network. A significant increase in neuron excitability was underlined in the presence of tumor cells.

Recent works uncover the neuronal impact on high-grade pediatric glioma (pHGG) cells and their reciprocal interactions. The present work shows the development of an in vitro model co-culturing pHGG cells and glutamatergic neurons and recorded their electrophysiological interactions to mimic those interactivities.

Glutamaterjik Nöronların ve Pediatrik Yüksek Dereceli Glioma Hücrelerinin Mikroakışkan Cihazlara Ortak Kültürü Elektriksel Etkileşimleri Değerlendirmek İçin

Pediatric high-grade gliomas (pHGG) represent childhood and adolescent brain cancers that carry a rapid dismal prognosis. Since there is a need to ...

Flexible Organic Electronic Devices for Pulsed Electric Field Therapy of Glioblastoma

Glioblastoma is difficult to eradicate with standard oncology therapies due to its high degree of invasiveness. Bioelectric treatments based on pulsed electric fields (PEFs) are promising for the improvement of treatment efficiency. However, they rely on rigid electrodes that cause acute and chronic damage, especially in soft tissues such as the brain. In this work, flexible electronics were used to deliver PEFs to tumors and the biological response was evaluated with fluorescent microscopy. Interdigitated gold electrodes on a thin, transparent parylene-C substrate were coated with the conducting polymer PEDOT:PSS, resulting in a conformable and biocompatible device. The effects of PEFs on tumors and their microenvironment were examined using various biological models. First, monolayers of glioblastoma cells were cultured on top of the electrodes to investigate phenomena in vitro. As an intermediate step, an in ovo model was developed where engineered tumor spheroids were grafted in the embryonic membrane of a quail. Due to the absence of an immune system, this led to highly vascularized tumors. At this early stage of development, embryos have no immune system, and tumors are not recognized as foreign bodies. Thus, they can develop fast while developing their own vessels from the existing embryo vascular system, which represents a valuable 3D cancer model. Finally, flexible electrode delivery of PEFs was evaluated in a complete organism with a functional immune system, using a syngenic, orthograft (intracranial) mouse model. Tumor spheroids were grafted into the brain of transgenic multi-fluorescent mice prior to the implantation of flexible organic electrode devices. A sealed cranial window enabled multiphoton imaging of the tumor and its microenvironment during treatment with PEFs over a period of several weeks.

This work describes the development of flexible interdigitated electrodes for implementation in 3D brain tumor models, namely, in vitro culture, in ovo model, and in vivo murine model. The proposed method can be used to evaluate the effects of pulsed electric fields on tumors at different levels of complexity.

Glioblastoma'nın Darbeli Elektrik Alan Terapisi için Esnek Organik Elektronik Cihazlar

Glioblastoma is difficult to eradicate with standard oncology therapies due to its high degree of invasiveness. Bioelectric treatments based on pulsed ...

Generation and Culturing of High-Grade Serous Ovarian Cancer Patient-Derived Organoids

Organoids are 3D dynamic tumor models that can be grown successfully from patient-derived ovarian tumor tissue, ascites, or pleural fluid and aid in the discovery of novel therapeutics and predictive biomarkers for ovarian cancer. These models recapitulate clonal heterogeneity, the tumor microenvironment, and cell-cell and cell-matrix interactions. Additionally, they have been shown to match the primary tumor morphologically, cytologically, immunohistochemically, and genetically. Thus, organoids facilitate research on tumor cells and the tumor microenvironment and are superior to cell lines. The present protocol describes distinct methods to generate patient-derived ovarian cancer organoids from patient tumors, ascites, and pleural fluid samples with a higher than 97% success rate. The patient samples are separated into cellular suspensions by both mechanical and enzymatic digestion. The cells are then plated utilizing a basement membrane extract (BME) and are supported with optimized growth media containing supplements specific to the culturing of high-grade serous ovarian cancer (HGSOC). After forming initial organoids, the PDOs can sustain long-term culture, including passaging for expansion for subsequent experiments.

Patient-derived organoids (PDO) are a three-dimensional (3D) culture that can mimic the tumor environment in vitro. In high-grade serous ovarian cancer, PDOs represent a model to study novel biomarkers and therapeutics.

Yüksek Dereceli Seröz Yumurtalık Kanseri Hasta Kaynaklı Organoidlerin Üretimi ve Kültürlenmesi

Organoids are 3D dynamic tumor models that can be grown successfully from patient-derived ovarian tumor tissue, ascites, or pleural fluid and aid in the ...

Application of the En Bloc Concept Combined with Anatomic Resection in Laparoscopic Hepatectomy

Laparoscopic hepatectomy has been reported in many studies, and it is the mainstream method of liver resection. In some particular cases, such as when there are tumors adjacent to the cystic bed, surgeons cannot palpate the surgical margins through the laparoscopic approach, which leads to uncertainty about R0 resection. Conventionally, the gallbladder is resected first, and the hepatic lobes or segments are resected second. However, tumor tissues can be disseminated in the above cases. To address this issue, based on the recognition of the porta hepatis and intrahepatic anatomy, we propose a unique approach to hepatectomy combined with gallbladder resection by en bloc anatomic resection in situ. Firstly, after dissecting the cystic duct, without cutting the gallbladder primarily, the porta hepatis is pre-occluded by the single lumen ureter; secondly, the left hepatic pedicle is made free by the gap of the Laennec membrane and Hilar plate; thirdly, the assistant is asked to drag the fundus of the gallbladder, and the liver parenchyma tissue is resected using a harmonic scalpel along the ischemia line on the liver surface and intraoperative ultrasound. The whole middle hepatic vein (MHV) and its tributaries appear completely; lastly, the left hepatic vein (LHV) is disconnected, and the specimen is taken out from the abdominal cavity. The tumor, gallbladder, and other surrounding tissues are resected en bloc, which meets the tumor-free criterion, and a wide incisal margin and R0 resection are achieved. Therefore, the laparoscopic hepatectomy with the combination of the en bloc concept and anatomic resection is a safe, effective, and radical method with low postoperative recurrence and metastasis.

Numerous studies have demonstrated the advantages of anatomic resection. Nonetheless, whether anatomic resection can increase R0 resection rates remains controversial. Consequently, the present study describes an innovative procedure involving the en bloc concept combined with anatomic resection in laparoscopic hepatectomy, which can reduce postoperative recurrence and metastasis.

Laparoskopik Hepatektomide Anatomik Rezeksiyon ile Kombine En Blok Kavramının Uygulanması

Laparoscopic hepatectomy has been reported in many studies, and it is the mainstream method of liver resection. In some particular cases, such as when ...