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      ENDCELL-Seud: a Delphi protocol to harmonise methods in endometrial cell culturing

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          Abstract

          In vitro

          culturing of endometrial cells obtained from the uterine mucosa or ectopic sites is used to study molecular and cellular signalling relevant to physiologic and pathologic reproductive conditions. However, the lack of consensus on standard operating procedures for deriving, characterising and maintaining primary cells in two- or three-dimensional cultures from eutopic or ectopic endometrium may be hindering progress in this area of research. Guidance for unbiased in vitro research methodologies in the field of reproductive science remains essential to increase confidence in the reliability of in vitro models. We present herein the protocol for a Delphi process to develop a consensus on in vitro methodologies using endometrial cells (ENDCELL-Seud Project). A steering committee composed of leading scientists will select critical methodologies, topics and items that need to be harmonised and that will be included in a survey. An enlarged panel of experts (ENDCELL-Seud Working Group) will be invited to participate in the survey and provide their ratings to the items to be harmonised. According to Delphi, an iterative investigation method will be adopted. Recommended measures will be finalised by the steering committee. The study received full ethical approval from the Ethical Committee of the Maastricht University (ref. FHML-REC/2021/103). The study findings will be available in both peer-reviewed articles and will also be disseminated to appropriate audiences at relevant conferences.

          Lay summary

          Patient-derived cells cultured in the lab are simple and cost-effective methods used to study biological and dysfunctional or disease processes. These tools are frequently used in the field of reproductive medicine. However, the lack of clear recommendations and standardised methodology to guide the laboratory work of researchers can produce results that are not always reproducible and sometimes are incorrect. To remedy this situation, we define here a method to ascertain if researchers who routinely culture cells in the lab agree or disagree on the optimal laboratory techniques. This method will be used to make recommendations for future researchers working in the field of reproductive biology to reproducibly culture endometrial cells in the laboratory.

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          Most cited references29

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          The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments.

          Currently, a lack of consensus exists on how best to perform and interpret quantitative real-time PCR (qPCR) experiments. The problem is exacerbated by a lack of sufficient experimental detail in many publications, which impedes a reader's ability to evaluate critically the quality of the results presented or to repeat the experiments. The Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines target the reliability of results to help ensure the integrity of the scientific literature, promote consistency between laboratories, and increase experimental transparency. MIQE is a set of guidelines that describe the minimum information necessary for evaluating qPCR experiments. Included is a checklist to accompany the initial submission of a manuscript to the publisher. By providing all relevant experimental conditions and assay characteristics, reviewers can assess the validity of the protocols used. Full disclosure of all reagents, sequences, and analysis methods is necessary to enable other investigators to reproduce results. MIQE details should be published either in abbreviated form or as an online supplement. Following these guidelines will encourage better experimental practice, allowing more reliable and unequivocal interpretation of qPCR results.
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            The COMET Handbook: version 1.0

            The selection of appropriate outcomes is crucial when designing clinical trials in order to compare the effects of different interventions directly. For the findings to influence policy and practice, the outcomes need to be relevant and important to key stakeholders including patients and the public, health care professionals and others making decisions about health care. It is now widely acknowledged that insufficient attention has been paid to the choice of outcomes measured in clinical trials. Researchers are increasingly addressing this issue through the development and use of a core outcome set, an agreed standardised collection of outcomes which should be measured and reported, as a minimum, in all trials for a specific clinical area. Accumulating work in this area has identified the need for guidance on the development, implementation, evaluation and updating of core outcome sets. This Handbook, developed by the COMET Initiative, brings together current thinking and methodological research regarding those issues. We recommend a four-step process to develop a core outcome set. The aim is to update the contents of the Handbook as further research is identified. Electronic supplementary material The online version of this article (doi:10.1186/s13063-017-1978-4) contains supplementary material, which is available to authorized users.
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              Stability of response characteristics of a Delphi panel: application of bootstrap data expansion

              Background Delphi surveys with panels of experts in a particular area of interest have been widely utilized in the fields of clinical medicine, nursing practice, medical education and healthcare services. Despite this wide applicability of the Delphi methodology, there is no clear identification of what constitutes a sufficient number of Delphi survey participants to ensure stability of results. Methods The study analyzed the response characteristics from the first round of a Delphi survey conducted with 23 experts in healthcare quality and patient safety. The panel members had similar training and subject matter understanding of the Malcolm Baldrige Criteria for Performance Excellence in Healthcare. The raw data from the first round sampling, which usually contains the largest diversity of responses, were augmented via bootstrap sampling to obtain computer-generated results for two larger samples obtained by sampling with replacement. Response characteristics (mean, trimmed mean, standard deviation and 95% confidence intervals) for 54 survey items were compared for the responses of the 23 actual study participants and two computer-generated samples of 1000 and 2000 resampling iterations. Results The results from this study indicate that the response characteristics of a small expert panel in a well-defined knowledge area are stable in light of augmented sampling. Conclusion Panels of similarly trained experts (who possess a general understanding in the field of interest) provide effective and reliable utilization of a small sample from a limited number of experts in a field of study to develop reliable criteria that inform judgment and support effective decision-making.
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                Author and article information

                Journal
                Reprod Fertil
                Reprod Fertil
                raf
                Reproduction & Fertility
                Bioscientifica Ltd (Bristol )
                2633-8386
                28 July 2022
                01 July 2022
                : 3
                : 3
                : G1-G8
                Affiliations
                [1 ]Department Obstetrics and Gynecology , GROW – School for Oncology and Reproduction, Maastricht University, Maastricht, The Netherlands
                [2 ]Shanghai Ob/Gyn Hospital , Fudan University, Shanghai, China
                [3 ]Division of Biomedical Sciences , Warwick Medical School University of Warwick, Coventry, UK
                [4 ]Department of Obstetrics , Gynecology & Reproductive Biology, Michigan State University, Grand Rapids, Michigan, USA
                [5 ]The Ritchie Centre , Hudson Institute of Medical Research and Department of Obstetrics and Gynecology, Monash University, Melbourne, Australia
                [6 ]Department of Instructive Biomaterials Engineering , MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht University, Maastricht, The Netherlands
                [7 ]Department of Gynecology and Obstetrics , Münster University Hospital, Münster, Germany
                [8 ]MIT , Cambridge, Massachusetts, USA
                [9 ]Yale University , New Haven, Connecticut, USA
                [10 ]University at Buffalo , Buffalo, New York, USA
                [11 ]University of Leuven , Leuven, Belgium
                [12 ]Université Paris Cité , Assistance Publique Hôpitaux de Paris, Centre Hospitalier Universitaire Cochin, Department of Gynecology Obstetrics II and Reproductive Medicine, Paris, France
                [13 ]Peking University People’s Hospital , Beijing, China
                [14 ]Department of Obstetrics and Gynecology , The Clinical and Translational Research Center, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan
                [15 ]Fondazione IRCCS Ca’ Granda , Ospedale Maggiore Policlinico, Milano, Italy
                Author notes
                Correspondence should be addressed to A Romano: a.romano@ 123456maastrichtuniversity.nl
                Author information
                http://orcid.org/0000-0002-5900-6883
                http://orcid.org/0000-0002-8511-7624
                http://orcid.org/0000-0002-9713-9134
                Article
                RAF-22-0041
                10.1530/RAF-22-0041
                9422235
                35972317
                63578b98-b7ae-4b64-a6bb-68a09860583c
                © The authors

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 29 June 2022
                : 28 July 2022
                Categories
                Guideline

                primary endometrial cells, in vitro culturing,protocol harmonisation,delphi method

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