Methods and Protocols最新文献_第7页

Development and Validation of Micro-Azocasein Assay for Quantifying Bromelain. 用于定量菠萝蛋白酶的微淀粉酶测定法的开发与验证

IF 2.4

Methods and Protocols Pub Date : 2024-03-15 DOI: 10.3390/mps7020025

Krishna Pillai, Javed Akhter, Ahmed H Mekkawy, Sarah J Valle, David L Morris

{"title":"Development and Validation of Micro-Azocasein Assay for Quantifying Bromelain.","authors":"Krishna Pillai, Javed Akhter, Ahmed H Mekkawy, Sarah J Valle, David L Morris","doi":"10.3390/mps7020025","DOIUrl":"10.3390/mps7020025","url":null,"abstract":"The proteolytic activity of enzymes may be evaluated by a colorimetric method with azocasein. Hence, we developed a micro-assay to quantify bromelain using azocasein. A total of 250 µL of 1.0% azocasein in dH2O was added to 250 µL of test solution, vortexed and incubated at ambient room temperature/30 min. The reaction was terminated with 1500 µL of 5% trichloroacetic acid, vortexed and centrifuged. A total of 150 µL of 0.5M NaOH was added to 150 µL of supernatant in triplicates, and absorbance was recorded at 410 nm. The linearity of the calibration curve was tested with 200-800 µg/mL serial dilutions. The detection limit, precision, accuracy, and robustness were tested along with the substrate enzyme reaction time and solvent matrix effect. Good linearity was seen with serially diluted 200 µg/mL bromelain. The limit of quantification and limit of detection were 5.412 and 16.4 µg/mL, respectively. Intra-day and inter-day analyses showed a relative standard deviation below 2.0%. The assay was robust when tested over 400-450 nm wavelengths. The assays performed using dH2O or PBS diluents indicated a higher sensitivity in dH2O. The proteolytic activity of bromelain was enhanced with L-cysteine or N-acetylcysteine. Hence, this micro-azocasein assay is reliable for quantifying bromelain.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10961761/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140207250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effects on Quality of Life of a Telemonitoring Platform amongst Patients with Cancer (EQUALITE): A Randomized Trial Protocol. 远程监控平台对癌症患者生活质量的影响（EQUALITE）：随机试验方案》。

IF 2.4

Methods and Protocols Pub Date : 2024-03-15 DOI: 10.3390/mps7020024

Felipe Martínez, Carla Taramasco, Manuel Espinoza, Johanna Acevedo, Carolina Goic, Bruno Nervi

{"title":"Effects on Quality of Life of a Telemonitoring Platform amongst Patients with Cancer (EQUALITE): A Randomized Trial Protocol.","authors":"Felipe Martínez, Carla Taramasco, Manuel Espinoza, Johanna Acevedo, Carolina Goic, Bruno Nervi","doi":"10.3390/mps7020024","DOIUrl":"10.3390/mps7020024","url":null,"abstract":"Cancer, a pervasive global health challenge, necessitates chemotherapy or radiotherapy treatments for many prevalent forms. However, traditional follow-up approaches encounter limitations, exacerbated by the recent COVID-19 pandemic. Consequently, telemonitoring has emerged as a promising solution, although its clinical implementation lacks comprehensive evidence. This report depicts the methodology of a randomized trial which aims to investigate whether leveraging a smartphone app called Contigo for disease monitoring enhances self-reported quality of life among patients with various solid cancers compared to standard care. Secondary objectives encompass evaluating the app's impact on depressive symptoms and assessing adherence to in-person appointments. Randomization will be performed independently using an allocation sequence that will be kept concealed from clinical investigators. Contigo offers two primary functions: monitoring cancer patients' progress and providing educational content to assist patients in managing common clinical situations related to their disease. The study will assess outcomes such as quality of life changes and depressive symptom development using validated scales, and adherence to in-person appointments. Specific scales include the EuroQol Group's EQ-5D questionnaire and the Patient Health Questionnaire (PHQ-9). We hypothesize that the use of Contigo will assist and empower patients receiving cancer treatment, which will translate to better quality of life scores and a reduced incidence of depressive symptoms. All analyses will be undertaken with the intention-to-treat principle by a statistician unaware of treatment allocation. This trial is registered in ClinicalTrials under the registration number NCT06086990.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10961787/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140207251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A Virtual Case Presentation Platform: Protocol Study. 虚拟病例展示平台：协议研究。

IF 2.4

Methods and Protocols Pub Date : 2024-03-08 DOI: 10.3390/mps7020023

Imad Alex Awada, Adina Magda Florea, Alexandru Scafa-Udriște

{"title":"A Virtual Case Presentation Platform: Protocol Study.","authors":"Imad Alex Awada, Adina Magda Florea, Alexandru Scafa-Udriște","doi":"10.3390/mps7020023","DOIUrl":"10.3390/mps7020023","url":null,"abstract":"Gaining practical experience is indispensable for medical students. Therefore, when medical students were prevented access to hospitals during the COVID-19 pandemic in Romania, there was an urgent need to find a solution that would allow medical students to develop the skills they would usually develop in hospitals but without the need to be physically present in a hospital. This was the reason behind the idea of developing a Virtual Case Presentation Platform. The platform offers the possibility for medical students to reproduce virtually, in clinically valid scenarios, the diagnostic process and treatment recommendation, as well as the interactions with patients that usually take place in hospitals using natural language through speech and text. On the platform, the students receive valuable feedback from the professors about their performance. In order to reproduce the whole targeted experience for students, without missing anything, before starting the development of the platform, it was mandatory to identify and understand all the aspects that should be covered by the platform. The proposed platform covers the different aspects that have been identified for the diagnostic process and treatment recommendation. It enables medical students to develop essential skills for their future careers as doctors.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10961792/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140207249","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Isolation and Culture of Primary Human Dental Pulp Cells—A Description of Technical and Methodological Steps to Maximise Predictability and Yield 原代人类牙髓细胞的分离和培养--最大限度提高可预测性和产量的技术和方法步骤说明

IF 2.4

Methods and Protocols Pub Date : 2024-03-01 DOI: 10.3390/mps7020022

Michaela Kearney, David E McReynolds, H. Duncan

{"title":"Isolation and Culture of Primary Human Dental Pulp Cells—A Description of Technical and Methodological Steps to Maximise Predictability and Yield","authors":"Michaela Kearney, David E McReynolds, H. Duncan","doi":"10.3390/mps7020022","DOIUrl":"https://doi.org/10.3390/mps7020022","url":null,"abstract":"The dental pulp has critical functions in tooth development as well as an ongoing role in promoting and maintaining the vitality of teeth. In particular, its regenerative ability allows dental tissues to be restored following damage caused by traumatic injury or caries. Regenerative endodontic procedures aim to utilise these processes to stimulate dental pulp repair in a minimally invasive manner and reduce the need for more invasive procedures such as root canal treatment. Dental pulp is a source of dental pulp cells (DPCs), which has a subpopulation of dental pulp stem cells (DPSCs), which are attractive for use in regenerative medicine due to their high proliferation rate, ability to differentiate into multiple cell types, and their preserved vitality following cryopreservation. The development of next-generation clinical therapeutics that maximise the potential of dental pulp relies on strong empirical evidence arising from in vitro experimentation. Here, we describe a modified method for the efficient isolation of primary human DPCs from sound third molar teeth for culture using an explant outgrowth method on basement membrane-coated flasks, as well as using high-resolution macro-photography to illustrate the methods. Critically, steps are taken to minimise potential physical and mechanical trauma to the cells and maximise yield. Human DPCs cultured using this method can be further expanded in cell culture flasks to facilitate their use in various in vitro experimental procedures.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140086551","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Acceptability, Feasibility, and Effectiveness of a Worksite Intervention to Lower Cardiometabolic Risk in South Africa: Protocol 南非降低心脏代谢风险的工作场所干预措施的可接受性、可行性和有效性：方案

IF 2.4

Methods and Protocols Pub Date : 2024-03-01 DOI: 10.3390/mps7020021

E. Singh, A. Naicker, Shivneta Singh

{"title":"Acceptability, Feasibility, and Effectiveness of a Worksite Intervention to Lower Cardiometabolic Risk in South Africa: Protocol","authors":"E. Singh, A. Naicker, Shivneta Singh","doi":"10.3390/mps7020021","DOIUrl":"https://doi.org/10.3390/mps7020021","url":null,"abstract":"As an important way to translate cardiovascular disease prevention efforts, worksite intervention programs can be used to effectively facilitate healthy food choices, health education, and social support among employees, in a targeted approach to improve health outcomes and physical activity levels of employees. In this study, the effectiveness of a canteen and a behavioral intervention on cardiometabolic risk among prediabetic and prehypertensive employees at two multinational worksites in South Africa will be measured. This two-arm randomized controlled trial (RCT) will be structured to provide a six-week intervention at two multinational companies spread across eight worksites and will include a canteen and behavioral arm (CB) and a canteen only (CO) arm. Participants who were either prediabetic or prehypertensive completed the baseline assessments, which included anthropometry, a demographic and lifestyle survey, the global physical activity questionnaire (GPAQ) and the 24 h food recall. Participants were randomized into the CO and the canteen and CB intervention groups. The CO group received six weeks of canteen intervention [changes to enable a healthy food environment], while the CB group received six weeks of canteen intervention along with a behavioral intervention. The behavioral intervention included an intense six-week lifestyle program aligned to the Diabetes Prevention Program (DPP). This study will assess the added benefit of environmental-level changes aimed at lowering cardiometabolic risk in a low–middle-income country (LMIC) and has the potential for scale-up to other worksites in South Africa and globally.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140090797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A Simple and Rapid Protocol for the Isolation of Murine Bone Marrow Suitable for the Differentiation of Dendritic Cells. 分离适合树突状细胞分化的小鼠骨髓的简单快速方案

IF 2.4

Methods and Protocols Pub Date : 2024-02-27 DOI: 10.3390/mps7020020

Runqiu Song, Mariam Bafit, Kirsteen M Tullett, Peck Szee Tan, Mireille H Lahoud, Meredith O'Keeffe, Anthony W Purcell, Asolina Braun

{"title":"A Simple and Rapid Protocol for the Isolation of Murine Bone Marrow Suitable for the Differentiation of Dendritic Cells.","authors":"Runqiu Song, Mariam Bafit, Kirsteen M Tullett, Peck Szee Tan, Mireille H Lahoud, Meredith O'Keeffe, Anthony W Purcell, Asolina Braun","doi":"10.3390/mps7020020","DOIUrl":"10.3390/mps7020020","url":null,"abstract":"The generation of bone-marrow-derived dendritic cells is a widely used approach in immunological research to study antigen processing and presentation, as well as T-cell activation responses. However, the initial step of isolating the bone marrow can be time-consuming, especially when larger numbers of precursor cells are required. Here, we assessed whether an accelerated bone marrow isolation method using centrifugation is suitable for the differentiation of FMS-like tyrosine kinase 3 ligand-driven dendritic cells. Compared to the conventional flushing method, the centrifugation-based isolation method resulted in a similar bone marrow cell yield on Day 0, increased cell numbers by Day 8, similar proportions of dendritic cell subsets, and consequently a higher number of type 1 conventional dendritic cells (cDC1) from the culture. Although the primary purpose of this method of optimization was to improve experimental efficiency and increase the output of cDC1s, the protocol is also compatible with the differentiation of other dendritic cell subsets such as cDC2 and plasmacytoid dendritic cells, with an improved output cell count and a consistent phenotype.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10961764/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140207248","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Scale-Up of the Fermentation Process for the Production and Purification of Serratiopeptidase Using Silkworm Pupae as a Substrate. 利用蚕蛹作为底物生产和纯化塞拉提肽酶的发酵工艺的规模化。

IF 2.4

Methods and Protocols Pub Date : 2024-02-25 DOI: 10.3390/mps7020019

Jhon Jairo Melchor-Moncada, Alejandra García-Barco, Augusto Zuluaga-Vélez, Luz Angela Veloza, Juan Carlos Sepúlveda-Arias

{"title":"Scale-Up of the Fermentation Process for the Production and Purification of Serratiopeptidase Using Silkworm Pupae as a Substrate.","authors":"Jhon Jairo Melchor-Moncada, Alejandra García-Barco, Augusto Zuluaga-Vélez, Luz Angela Veloza, Juan Carlos Sepúlveda-Arias","doi":"10.3390/mps7020019","DOIUrl":"10.3390/mps7020019","url":null,"abstract":"Serratiopeptidase, a bacterial metalloprotease known for its pain-relieving and anti-inflammatory properties, can be produced through fermentation with S. marcescens. This study aimed to identify key factors related to nutrient composition and physicochemical conditions for production in Erlenmeyer flasks and to scale up the mixture to a bioreactor to obtain the maximum proteolytic activity. A Plackett-Burman design was used to determine whether the presence of silkworm pupae (at 1.5%) was a significant parameter for serratiopeptidase production. Along with the variables pH, temperature, and time, they were optimized using a Taguchi experimental design, resulting in values of 7, 25 °C, and 36 h, respectively. Scaling up with a kLa of 25.45 ± 3.12 h-1 showed the highest serratiopeptidase production at 24 h. A factorial design was used for ultrafiltration, resulting in an LMH (liters per square meter per hour) of 960 L/m2h, a TMP (transmembrane pressure) of 15 psi, and a concentration factor of five, with a specific activity of 24,325.81 ± 1515.69 U/mg. Afterward, the retentate was purified using strong anion exchange chromatography and ultrafiltration, yielding a 19.94 ± 3.07% recovery and a purification factor of 1.59 ± 0.31. In conclusion, waste from the sericulture industry can be used for serratiopeptidase production.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10961818/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140207252","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Housing European Ground Squirrels (Spermophilus citellus) for an Ex Situ Conservation Program. 欧洲地松鼠（Spermophilus citellus）原生境保护计划的饲养场。

IF 2.4

Methods and Protocols Pub Date : 2024-02-20 DOI: 10.3390/mps7020018

Boróka Bárdos, Vilmos Altbacker, Henrietta Kinga Török, István Nagy

{"title":"Housing European Ground Squirrels (Spermophilus citellus) for an Ex Situ Conservation Program.","authors":"Boróka Bárdos, Vilmos Altbacker, Henrietta Kinga Török, István Nagy","doi":"10.3390/mps7020018","DOIUrl":"10.3390/mps7020018","url":null,"abstract":"European ground squirrel (Spermophilus citellus) populations have declined precipitously over the last 70 years. Its protection cannot be ensured solely by protecting its habitat; it is also necessary to protect the animals ex situ. In our study, within a European ground squirrel species protection program, we examined two elements of indoor housing technology. Knowledge of the animals' needs is essential for captive housing and breeding success, so in our tests, the animals could freely choose both nest-building materials and feed. In the nest material preference test, the animals could choose from three materials with different structures: paper, Lignocel and hay. In the feed preference test, the animals could also choose from three types of feed: commercial rabbit feed, complete rabbit feed and a natural feed mixture. The first two feeds were in granulated format, and the third was a grain feed mix. Among the nesting materials, they preferred hay, which allowed them to build better-quality nests. Among the feeds, they preferred the grain feed mix, the composition closest to their natural feed, and it was the only one that contained animal protein. Our results contribute to the successful maintenance and breeding the European ground squirrel in captivity.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10885073/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139931972","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Crafting a Rigorous, Clinically Relevant Large Animal Model of Chronic Myocardial Ischemia: What Have We Learned in 20 Years? 建立严格的、与临床相关的慢性心肌缺血大型动物模型：20 年来我们学到了什么？

IF 2.3

Methods and Protocols Pub Date : 2024-02-19 DOI: 10.3390/mps7010017

Christopher R Stone, Dwight D Harris, Mark Broadwin, Meghamsh Kanuparthy, Sharif A Sabe, Cynthia Xu, Jun Feng, M Ruhul Abid, Frank W Sellke

{"title":"Crafting a Rigorous, Clinically Relevant Large Animal Model of Chronic Myocardial Ischemia: What Have We Learned in 20 Years?","authors":"Christopher R Stone, Dwight D Harris, Mark Broadwin, Meghamsh Kanuparthy, Sharif A Sabe, Cynthia Xu, Jun Feng, M Ruhul Abid, Frank W Sellke","doi":"10.3390/mps7010017","DOIUrl":"10.3390/mps7010017","url":null,"abstract":"The past several decades have borne witness to several breakthroughs and paradigm shifts within the field of cardiovascular medicine, but one component that has remained constant throughout this time is the need for accurate animal models for the refinement and elaboration of the hypotheses and therapies crucial to our capacity to combat human disease. Numerous sophisticated and high-throughput molecular strategies have emerged, including rational drug design and the multi-omics approaches that allow extensive characterization of the host response to disease states and their prospective resolutions, but these technologies all require grounding within a faithful representation of their clinical context. Over this period, our lab has exhaustively tested, progressively refined, and extensively contributed to cardiovascular discovery on the basis of one such faithful representation. It is the purpose of this paper to review our porcine model of chronic myocardial ischemia using ameroid constriction and the subsequent myriad of physiological and molecular-biological insights it has allowed our lab to attain and describe. We hope that, by depicting our methods and the insight they have yielded clearly and completely-drawing for this purpose on comprehensive videographic illustration-other research teams will be empowered to carry our work forward, drawing on our experience to refine their own investigations into the pathogenesis and eradication of cardiovascular disease.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10891802/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139931970","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Assessment of the Integrity and Function of Human Term Placental Explants in Short-Term Culture 评估短期培养的人类末期胎盘外胚层的完整性和功能

IF 2.4

Methods and Protocols Pub Date : 2024-02-15 DOI: 10.3390/mps7010016

Carolina López-Guzmán, Ana María García, Paula Marín, Ana María Vásquez

{"title":"Assessment of the Integrity and Function of Human Term Placental Explants in Short-Term Culture","authors":"Carolina López-Guzmán, Ana María García, Paula Marín, Ana María Vásquez","doi":"10.3390/mps7010016","DOIUrl":"https://doi.org/10.3390/mps7010016","url":null,"abstract":"Human placental explants (HPEs) culture has generated significant interest as a valuable in vitro model for studying tissue functions in response to adverse conditions, such as fluctuations in oxygen levels, nutrient availability, exposure to pathogenic microorganisms, and toxic compounds. HPEs offers the advantage of replicating the intricate microenvironment and cell-to-cell communication involved in this critical and transient organ. Although HPEs culture conditions have been extensively discussed, a protocol for assessing the viability and function of HPEs during short-term culture has not been previously outlined. In this study, we have developed a short-term HPEs culture protocol, specifically up to 72 h, and have employed quantitative, semi-quantitative, and qualitative analyses to evaluate tissue viability and function over time. Under our standardized conditions, placental villi explants began to regain their structural properties (the integrity of the trophoblast and villous stroma) and the functionality of the HPEs (production of angiogenic, endocrine, and immunological factors) starting from 48 h of culture. This restoration ensures a suitable environment for several applications. The data presented here can be highly valuable for laboratories aiming to implement an HPEs model, whether in the process of standardization or seeking to enhance and optimize working conditions and timing with placental tissue.","PeriodicalId":18715,"journal":{"name":"Methods and Protocols","volume":null,"pages":null},"PeriodicalIF":2.4,"publicationDate":"2024-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139834991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0