Good practice recommendations for the use of time-lapse technology†

STUDY QUESTION: What recommendations can be provided on the approach to and use of time-lapse technology (TLT) in an IVF laboratory?SUMMARY ANSWER: The present ESHRE document provides 11 recommendations on how to introduce TLT in the IVF laboratory. WHAT IS KNOWN ALREADY: Studies have been published on the use of TLT in clinical embryology. However, a systematic assessmentof how to approach and introduce this technology is currently missing.STUDY DESIGN, SIZE, DURATION: A working group of members of the Steering Committee of the ESHRE Special Interest Group in Embryology and selected ESHRE members was formed in order to write recommendations on the practical aspects of TLT for the IVF laboratory.PARTICIPANTS/MATERIALS, SETTING, METHODS: The working group included 11 members of different nationalities with internationally recognized experience in clinical embryology and basic science embryology, in addition to TLT. This document is developed according to the manual for development of ESHRE recommendations for good practice. Where possible, the statements are supported by studies retrieved from a PUBMED literature search on ‘time-lapse’ and ART.MAIN RESULTS AND THE ROLE OF CHANCE: A clear clinical benefit of the use of TLT, i.e. an increase in IVF success rates, remains to be proven. Meanwhile, TLT systems are being introduced in IVF laboratories. The working group listed 11 recommendations on what to do before introducing TLT in the lab. These statements include an assessment of the pros and cons of acquiring a TLT system, selection of relevant morphokinetic parameters, selection of an appropriate TLT system with technical and customer support, development of an internal checklist and education of staff. All these aspects are explained further here, based on the current literature and expert opinion.LIMITATIONS, REASONS FOR CAUTION: Owing to the limited evidence available, recommendations are mostly based on clinical and technical expertise. The paper provides technical advice, but leaves any decision on whether or not to use TLT to the individual centres.WIDER IMPLICATIONS OF THE FINDINGS: This document is expected to have a significant impact on future developments of clinical embryology, considering the increasing role and impact of TLT

Unilateral Renal Ischemia-Reperfusion as a Robust Model for Acute to Chronic Kidney Injury in Mice.

Acute kidney injury (AKI) is an underestimated, yet important risk factor for development of chronic kidney disease (CKD). Even after initial total recovery of renal function, some patients develop progressive and persistent deterioration of renal function and these patients are more likely to progress to end-stage renal disease (ESRD). Animal models are indispensable for unravelling the mechanisms underlying this progression towards CKD and ESRD and for the development of new therapeutic strategies in its prevention or treatment. Ischemia (i.e. hypoperfusion after surgery, bleeding, dehydration, shock, or sepsis) is a major aetiology in human AKI, yet unilateral ischemia-reperfusion is a rarely used animal model for research on CKD and fibrosis. Here, we demonstrate in C57Bl/6J mice, by both histology and gene expression, that unilateral ischemia-reperfusion without contralateral nephrectomy is a very robust model to study the progression from acute renal injury to long-term tubulo-interstitial fibrosis, i.e. the histopathological hallmark of CKD. Furthermore, we report that the extent of renal fibrosis, in terms of Col I, TGFβ, CCN2 and CCN3 expression and collagen I immunostaining, increases with increasing body temperature during ischemia and ischemia-time. Thus, varying these two main determinants of ischemic injury allows tuning the extent of the long-term fibrotic outcome in this model. Finally, in order to cover the whole practical finesse of ischemia-reperfusion and allow model and data transfer, we provide a referenced overview on crucial technical issues (incl. anaesthesia, analgesia, and pre- and post-operative care) with the specific aim of putting starters in the right direction of implementing ischemia in their research and stimulate them, as well as the community, to have a critical view on ischemic literature data

Photos of Masson’s stained slides of ischemic kidney tissue.

<p>The images shown are representative of the group. Masson’s stain showed prominent renal damage and severe loss of structure, with necrotic cells (arrowhead), casts or intraluminal debris (arrow), inflammatory infiltration and fibrosis (*). Blue stain represents extracellular matrix deposition (i.e. fibrosis). Magnification: 200x. <b>A:</b> Effect of body temperature on long-term fibrotic outcome 12 weeks after UIRI. <b>B:</b> Effect of ischemia time on long-term fibrotic outcome 6 and 12 weeks after UIRI.</p

Collagen I immunostaining in the ischemic kidneys.

<p>*: p<0.05, °: p<0.05 vs. Sham. <b>A:</b> Effect of body temperature on long-term collagen I deposition in the ischemic kidney, 12 weeks after UIRI (magnification: 100x). <b>B:</b> Effect of ischemia time on long-term collagen I deposition in the ischemic kidney, 6 and 12 weeks after UIRI (magnification: 100x). <b>C:</b> UIRI was performed for 30 minutes at 37°C (n = 5), 36°C (n = 4), 35°C (n = 10) or 34°C (n = 5) and animals were euthanized 12 weeks after UIRI. Collagen I deposition seems to be dependent on body temperature during ischemia: more collagen I deposition after UIRI at higher body temperatures. <b>D:</b> UIRI was performed for 30, 21 or 18 minutes at 36°C and animals were euthanized 6 weeks (resp. n = 5, n = 12, n = 6) and 12 weeks (resp. n = 4, n = 5, n = 10) after UIRI. Collagen I deposition seems to be ischemia time-dependent: more collagen I deposition after longer ischemia times. The bars are the means ± s.d. The data were analysed using a two-tailed Mann-Whitney U test.</p

Relative quantification of long-term IRI-induced expression of fibrosis-related genes.

<p>Core body temperature during ischemia determines degree of long-term fibrotic outcome. *: p<0.05, °: p<0.05 vs. Sham. UIRI was performed for 30 minutes at 37°C (n = 5), 36°C (n = 4), 35°C (n = 10) or 34°C (n = 5) and animals were euthanized 12 weeks after UIRI. Twelve weeks after UIRI, a significant increase in gene expression of fibrosis-related genes <i>Col I</i>, <i>TGFβ</i>, <i>CCN2</i> and <i>CCN3</i> was observed in renal cortex tissue in all core body temperature conditions tested. The expression of these genes is also temperature-dependent: higher expression with higher temperature during ischemia. The bars are the means ± s.d. The data were analysed using a two-tailed Mann-Whitney U test.</p

Relative quantification of long-term IRI-induced expression of tubular injury and inflammatory markers.

<p>*: p<0.05, °: p<0.05 vs. Sham, #: p<0.05 vs. week 6. UIRI was performed for 30, 21 or 18 minutes at 36°C and animals were euthanized 6 weeks (resp. n = 5, n = 12, n = 6) and 12 weeks (resp. n = 4, n = 5, n = 10) after UIRI. <b>A:</b> Six weeks after 30, 21 and 18 minutes of UIRI, a significant increase in gene expression of tubular injury markers <i>Havcr1</i> (KIM-1) and <i>Lcn2</i> (NGAL) was observed. At 12 weeks after UIRI, upregulation of these markers is ischemia-time dependent, with higher upregulation with longer ischemia times. <b>B:</b> Six weeks after 30, 21 and 18 minutes of UIRI, a significant increase in gene expression of inflammatory cytokines <i>TNFα</i> and <i>IL-6</i> was observed. In addition, short ischemia times, i.e. 18 minutes of UIRI, induced significantly lower gene expression of these markers. At 12 weeks after UIRI, upregulation of these inflammatory cytokines shows an ischemia-time dependent effect, with shorter ischemia times inducing less upregulation of gene expression of these inflammatory markers.</p

ESHRE guideline : medically assisted reproduction in patients with a viral infection/disease

STUDY QUESTION: What is the recommended management for medically assisted reproduction (MAR) in patients with a viral infection or disease, based on the best available evidence in the literature? SUMMARY ANSWER: The ESHRE guideline on MAR in patients with a viral infection/disease makes 78 recommendations on prevention of horizontal and vertical transmission before, during and after MAR, and the impact on its outcomes, and these also include recommendations regarding laboratory safety on the processing and storage of gametes and embryos testing positive for viral infections. WHAT IS KNOWN ALREADY: The development of new and improved anti-viral medications has resulted in improved life expectancy and quality of life for patients with viral infections/diseases. Patients of reproductive age are increasingly exploring their options for family creation. STUDY DESIGN, SIZE, DURATION: The guideline was developed according to the structured methodology for the development of ESHRE guidelines. After the formulation of nine key questions for six viruses (hepatitis B virus, hepatitis C virus, human immunodeficiency virus, human papilloma virus, human T-lymphotropic virus I/11 and Zika virus) by a group of experts, literature searches and assessments were performed. Papers published up to 2 November 2020 and written in English were included in the review. Evidence was analyzed by female, male or couple testing positive for the virus. PARTICIPANTS/MATERIALS, SETTING, METHODS: Based on the collected evidence, recommendations were formulated and discussed until consensus was reached within the guideline group. There were 61 key questions to be answered by the guideline development group (GDG), of which 12 were answered as narrative questions and 49 as PICO (Patient, Intervention, Comparison, Outcome) questions. A stakeholder review was organized after the finalization of the draft. The final version was approved by the GDG and the ESHRE Executive Committee. MAIN RESULTS AND THE ROLE OF CHANCE: This guideline aims to help providers meet a growing demand for guidance on the management of patients with a viral infection/disease presenting in the fertility clinic. The guideline makes 78 recommendations on prevention of viral transmission before and during MAR, and interventions to reduce/avoid vertical transmission to the newborn. Preferred MAR treatments and interventions are described together with the effect of viral infections on outcomes. The GDG formulated 44 evidence-based recommendations-of which 37 were formulated as strong recommendations and 7 as weak-33 good practice points (GPP) and one research only recommendation. Of the evidence-based recommendations, none were supported by high-quality evidence, two by moderate-quality evidence, 15 by low-quality evidence and 27 by very low-quality evidence. To support future research in the field of MAR in patients with a viral infection/disease, a list of research recommendations is provided. LIMITATIONS, REASONS FOR CAUTION: Most interventions included are not well-studied in patients with a viral infection/disease. For a large proportion of interventions, evidence was very limited and of very low quality. More evidence is required for these interventions, especially in the field of human papilloma virus (HPV). Such future studies may require the current recommendations to be revised. WIDER IMPLICATIONS OF THE FINDINGS: The guideline provides clinicians with clear advice on best practice in MAR for patients with a viral infection/disease, based on the best evidence currently available. In addition, a list of research recommendations is provided to stimulate further studies in the field

ESHRE guideline: ovarian stimulation for IVF/ICSI†

What is the recommended management of ovarian stimulation, based on the best available evidence in the literature