A novel method to efficiently differentiate human osteoclasts from blood-derived monocytes.

BACKGROUND: Osteoclasts are the tissue-specific macrophage population of the bone and unique in their bone-resorbing activity. Hence, they are fundamental for bone physiology in health and disease. However, efficient protocols for the isolation and study of primary human osteoclasts are scarce. In this study, we aimed to establish a protocol, which enables the efficient differentiation of functional human osteoclasts from monocytes. RESULTS: Human monocytes were isolated through a double-density gradient from donor blood. Compared to standard differentiation schemes in polystyrene cell culture dishes, the yield of multinuclear osteoclasts was significantly increased upon initial differentiation of monocytes to macrophages in fluorinated ethylene propylene (FEP) Teflon bags. This initial differentiation phase was then followed by the development of terminal osteoclasts by addition of Receptor Activator of NF-κB Ligand (RANKL). High concentrations of RANKL and Macrophage colony-stimulating factor (M-CSF) as well as an intermediate cell density further supported efficient cell differentiation. The generated cells were highly positive for CD45, CD14 as well as the osteoclast markers CD51/ITGAV and Cathepsin K/CTSK, thus identifying them as osteoclasts. The bone resorption of the osteoclasts was significantly increased when the cells were differentiated from macrophages derived from Teflon bags compared to macrophages derived from conventional cell culture plates. CONCLUSION: Our study has established a novel protocol for the isolation of primary human osteoclasts that improves osteoclastogenesis in comparison to the conventionally used cultivation approach.

PD-L1 on large extracellular vesicles is a predictive biomarker for therapy response in tissue PD-L1-low and -negative patients with non-small cell lung cancer.

Immunotherapy has revolutionized the treatment of patients with non-small cell lung cancer (NSCLC). High expression of tissue PD-L1 (tPD-L1) is currently the only approved biomarker for predicting treatment response. However, even tPD-L1 low (1-49%) and absent (<1%) patients might benefit from immunotherapy but, to date, there is no reliable biomarker, that can predict response in this particular patient subgroup. This study aimed to test whether tumour-associated extracellular vesicles (EVs) could fill this gap. Using NSCLC cell lines, we identified a panel of tumour-related antigens that were enriched on large EVs (lEVs) compared to smaller EVs. The levels of lEVs carrying these antigens were significantly elevated in plasma of NSCLC patients (n = 108) and discriminated them from controls (n = 77). Among the tested antigens, we focused on programmed cell death ligand 1 (PD-L1), which is a well-known direct target for immunotherapy. In plasma lEVs, PD-L1 was mainly found on a population of CD45- /CD62P+ lEVs and thus seemed to be associated with platelet-derived vesicles. Patients with high baseline levels of PD-L1+ lEVs in blood showed a significantly better response to immunotherapy and prolonged survival. This was particularly true in the subgroup of NSCLC patients with low or absent tPD-L1 expression, thus identifying PD-L1-positive lEVs in plasma as a novel predictive and prognostic marker for immunotherapy.

Corrigendum to "Geographic network effects to engage people in the energy transition: The case of PV in Switzerland" [Heliyon 9(7) (July 2023) e17800].

[This corrects the article DOI: 10.1016/j.heliyon.2023.e17800.].

Extracellular vesicle-associated tyrosine kinase-like orphan receptors ROR1 and ROR2 promote breast cancer progression.

BACKGROUND: Extracellular vesicles (EVs) harbor a plethora of different biomolecules, which they can transport across cells. In cancer, tumor-derived EVs thereby support the creation of a favorable tumor microenvironment. So far, EV uptake and cargo delivery into target cells have been regarded as the main mechanisms for the pro-tumoral function of EVs. To test this hypothesis, we investigated the fate of the oncogenic transmembrane Wnt tyrosine kinase-like orphan receptor 1 and 2 (ROR1, ROR2) delivered via distinct EV subpopulations to breast cancer cells and aimed to unravel their impact on tumor progression. METHODS: EVs were isolated by differential ultracentrifugation from cell culture supernatant as well as plasma samples from healthy individuals (n = 27) and breast cancer patients (n = 41). EVs were thoroughly characterized by electron microscopy, nanoparticle tracking analysis, immunoblot, and flow cytometry. ROR transfer to target cells was observed using microscopy-based assays and biodistribution experiments were conducted in syngeneic mice. EV impact on cancer cell migration and invasion was tested in functional assays. RESULTS: We observed that the supernatant of ROR-overexpressing cells was sufficient for transferring the receptors to ROR-negative cells. Analyzing the secretome of the ROR-overexpressing cells, we detected a high enrichment of ROR1/2 on large and small EVs, but not on large oncosomes. Interestingly, the majority of ROR-positive EVs remained attached to the target cell surface after 24 h of stimulation and was quickly removed by treatment with trypsin. Nonetheless, ROR-positive EVs increased migration and invasion of breast cancer cells, even after chemically inhibiting EV uptake, in dependence of RhoA downstream signaling. In vivo, ROR-depleted EVs tended to distribute less into organs prone for the formation of breast cancer metastases. ROR-positive EVs were also significantly elevated in the plasma of breast cancer patients and allowed to separate them from healthy controls. CONCLUSIONS: The oncogenic Wnt receptors ROR1/2 are transferred via EVs to the surface of ROR-negative cancer cells, in which they induce an aggressive phenotype supporting tumor progression. Video Abstract.

Geographic network effects to engage people in the energy transition: The case of PV in Switzerland.

With the energy transition and CO2 emissions reduction as a priority on the international policy agenda, governments worldwide are trying to engage the population in investing in renewable energies. In this paper, we study the role of information access and peer effects in the photovoltaics sector in the case of the Swiss canton of Vaud. Based on a representative survey of the population of two districts, Nyon and Jura-Nord Vaudois, we show that being a homeowner and knowing someone who has installed PV cells in your social group significantly increase the probability of putting up PV cells. A direct neighbourhood effect was found, meaning that if a neighbour has installed a PV cell, the probability of the inhabitants in the two case study regions installing one themselves increases significantly. Our results show that spatial proximity is an important factor in the transmission of information between peers. Besides pure geographical distance, additional aspects such as administrative boundaries, shared language or degree of urbanisation play a role in the way the information network is presented. These insights indicate that professional experts and neighbours are important points of reference in the decision to invest in PV, and that regional networks are key for the active spreading of information on renewable technologies. Thus, we recommend using these connections to actively promote PV.

Metabolic Changes in Blood-derived Extracellular Vesicles of Malnourished Breast Cancer Patients.

BACKGROUND/AIM: Cancer cachexia describes a multifactorial wasting syndrome marked by a metabolic imbalance leading to the loss of muscle and fat tissue. Extracellular vesicles (EV) provide unique insights into their parental cells' metabolism. The value of these vesicles as diagnostic tools in cancer cachexia has not been investigated so far. PATIENTS AND METHODS: A previously analyzed metabolomics dataset on large EV from breast cancer patients was used for analyzing the metabolomic changes in patients with malnutrition. Follow-up time was 6 months. The data were analyzed using fold change analysis, volcano plotting, receiver operator characteristic (ROC) analysis, pathway analysis, and survival analysis. RESULTS: In patients with weight loss, statistical analysis revealed an increase in lysophosphatidylcholines (lysoPC a C16:0, lysoPC a C18:0, lysoPC a C18:1, lysoPC a C18:2, lysoPC a C20:4), sphingomyelins (SM (OH) C22:2 and SM C18:1), and phosphatidylcholines (PC aa C24:0, PC ae C34:3). When combined, these metabolites are a good predictor for cachexia in ROC curve analysis (AUC of 0.970; 95%CI=0.920-1.000; p<0.0001). Pathway analysis revealed an involvement of metabolites in "choline metabolism in cancer" and "glycerophospholipid metabolism". CONCLUSION: Large EV reflect metabolic changes in cancer patients suffering from cancer cachexia. Metabolic changes at the time of drawing blood were associated with the weight status (stable vs. weight loss) six months later and thereby could have a predictive impact.

Obstacles and opportunities for reducing dwelling size to shrink the environmental footprint of housing: tenants' residential preferences and housing choice.

The environmental footprint of housing is greatly influenced by the size of a dwelling. Housing size is the result of households' dwelling selections; accordingly, it is critical to consider residential preferences and choices to inform efforts towards housing sustainability. This study aimed to understand tenants' preferences for and choices of housing size as one amongst several dwelling characteristics and identify obstacles and opportunities for reducing size in the light of promoting sustainable housing. We employed logistic regression models to analyse a survey with 878 Swiss tenants, and our results identify preference for large dwellings as a major obstacle for reducing dwelling size among affluent tenants. Conversely, tenants with lower income might be forced to move to a smaller dwelling due to financial constraints or attribute higher importance to the financial benefit of lower rents. However, financial disincentives along with substantial non-monetary costs of moving, such as the disruption of local bonds and the difficulty of finding a satisfactory dwelling, can outweigh the benefits of moving to a smaller dwelling. To overcome such obstacles, we suggest offering incentives and other facilitating measures for downsizing moves as well as ensuring an adequate supply of smaller dwellings capable of providing high living quality. We highlight the potential of studying housing functions to conceptualize dwellings fulfilling these requirements.

Superresolution microscopy localizes endogenous Dvl2 to Wnt signaling-responsive biomolecular condensates.

During organismal development, homeostasis, and disease, Dishevelled (Dvl) proteins act as key signaling factors in beta-catenin-dependent and beta-catenin-independent Wnt pathways. While their importance for signal transmission has been genetically demonstrated in many organisms, our mechanistic understanding is still limited. Previous studies using overexpressed proteins showed Dvl localization to large, punctate-like cytoplasmic structures that are dependent on its DIX domain. To study Dvl's role in Wnt signaling, we genome engineered an endogenously expressed Dvl2 protein tagged with an mEos3.2 fluorescent protein for superresolution imaging. First, we demonstrate the functionality and specificity of the fusion protein in beta-catenin-dependent and beta-catenin-independent signaling using multiple independent assays. We performed live-cell imaging of Dvl2 to analyze the dynamic formation of the supramolecular cytoplasmic Dvl2_mEos3.2 condensates. While overexpression of Dvl2_mEos3.2 mimics the previously reported formation of abundant large "puncta," supramolecular condensate formation at physiological protein levels is only observed in a subset of cells with approximately one per cell. We show that, in these condensates, Dvl2 colocalizes with Wnt pathway components at gamma-tubulin and CEP164-positive centrosomal structures and that the localization of Dvl2 to these condensates is Wnt dependent. Single-molecule localization microscopy using photoactivated localization microscopy (PALM) of mEos3.2 in combination with DNA-PAINT demonstrates the organization and repetitive patterns of these condensates in a cell cycle-dependent manner. Our results indicate that the localization of Dvl2 in supramolecular condensates is coordinated dynamically and dependent on cell state and Wnt signaling levels. Our study highlights the formation of endogenous and physiologically regulated biomolecular condensates in the Wnt pathways at single-molecule resolution.

High spatial resolution dataset of La Mobilière insurance customers.

We present the La Mobilière insurance customers dataset: a 12-year-long longitudinal collection of data on policies of customers of the Swiss insurance company La Mobilière. To preserve the privacy of La Mobilière customers, we propose the data aggregated at two geographical levels, based on the place of residence of the customer: postal areas and municipalities. For each geographical area, the data provides summary statistics on: (i) the demographic characteristics of the customer base, (ii) characteristics of vehicles insurance policies and (iii) characteristics of housing and building insurance policies. To assess the validity of the data, we investigate the temporal consistency of the data and the representativeness of La Mobilière customer base along several dimensions (total population, percentage of foreigners, etc.). We also show how the insurance data can reliably model the spatial patterns of socio-economic indicators at a high geographical resolution. We believe that the reuse of this data provides an opportunity for researchers to broaden the socio-economic characterization of Swiss areas beyond the use of official data sources.

Towards circular phosphorus: The need of inter- and transdisciplinary research to close the broken cycle.

Phosphorus (P) is an essential element to all living beings but also a finite resource. P-related problems center around broken P cycles from local to global scales. This paper presents outcomes from the 9th International Phosphorus Workshop (IPW9) held 2019 on how to move towards a sustainable P management. It is based on two sequential discussion rounds with all participants. Important progress was reported regarding the awareness of P as finite mineable resource, technologies to recycle P, and legislation towards a circular P economy. Yet, critical deficits were identified such as how to handle legacy P, how climate change may affect ecosystem P cycling, or working business models to up-scale existing recycling models. Workshop participants argued for more transdisciplinary networks to narrow a perceived science-practice/policy gap. While this gap may be smaller in reality as illustrated with a Swiss example, we formulate recommendations how to bridge this gap more effectively.

WNT11/ROR2 signaling is associated with tumor invasion and poor survival in breast cancer.

BACKGROUND: Breast cancer has been associated with activation of the WNT signaling pathway, although no driver mutations in WNT genes have been found yet. Instead, a high expression of the alternative WNT receptor ROR2 was observed, in particular in breast cancer brain metastases. However, its respective ligand and downstream signaling in this context remained unknown. METHODS: We modulated the expression of ROR2 in human breast cancer cells and characterized their gene and protein expression by RNA-Seq, qRT-PCR, immunoblots and reverse phase protein array (RPPA) combined with network analyses to understand the molecular basis of ROR2 signaling in breast cancer. Using co-immunoprecipitations, we verified the interaction of ROR2 with the identified ligand, WNT11. The functional consequences of WNT11/ROR2 signaling for tumor cell aggressiveness were assessed by microscopy, impedance sensing as well as viability and invasion assays. To evaluate the translational significance of our findings, we performed gene set enrichment, expression and survival analyses on human breast cancer brain metastases. RESULTS: We found ROR2 to be highly expressed in aggressive breast tumors and associated with worse metastasis-free survival. ROR2 overexpression induced a BRCAness-like phenotype in a cell-context specific manner and rendered cells resistant to PARP inhibition. High levels of ROR2 were furthermore associated with defects in cell morphology and cell-cell-contacts leading to increased tumor invasiveness. On a molecular level, ROR2 overexpression upregulated several non-canonical WNT ligands, in particular WNT11. Co-immunoprecipitation confirmed that WNT11 indeed interacts with the cysteine-rich domain of ROR2 and triggers its invasion-promoting signaling via RHO/ROCK. Knockdown of WNT11 reversed the pro-invasive phenotype and the cellular changes in ROR2-overexpressing cells. CONCLUSIONS: Taken together, our study revealed a novel auto-stimulatory loop in which ROR2 triggers the expression of its own ligand, WNT11, resulting in enhanced tumor invasion associated with breast cancer metastasis.

Metabolomic Profiling of Blood-Derived Microvesicles in Breast Cancer Patients.

Malignant cells differ from benign ones in their metabolome and it is largely unknown whether this difference is reflected in the metabolic profile of their microvesicles (MV), which are secreted into the blood of cancer patients. Here, they are present together with MV from the various blood and endothelial cells. Harvesting MV from 78 breast cancer patients (BC) and 30 controls, we characterized the whole blood MV metabolome using targeted and untargeted mass spectrometry. Especially (lyso)-phosphatidylcholines and sphingomyelins were detected in a relevant abundance. Eight metabolites showed a significant discriminatory power between BC and controls. High concentrations of lysoPCaC26:0 and PCaaC38:5 were associated with shorter overall survival. Comparing BC subtype-specific metabolome profiles, 24 metabolites were differentially expressed between luminal A and luminal B. Pathway analysis revealed alterations in the glycerophospholipid metabolism for the whole cancer cohort and in the ether lipid metabolism for the molecular subtype luminal B. Although this mixture of blood-derived MV contains only a minor number of tumor MV, a combination of metabolites was identified that distinguished between BC and controls as well as between molecular subtypes, and was predictive for overall survival. This suggests that these metabolites represent promising biomarkers and, moreover, that they may be functionally relevant for tumor progression.

Tenants' residential mobility in Switzerland: the role of housing functions.

The interaction between residential preferences and dwellings is a complex system whose function thus far remains insufficiently explored. In this paper, we investigate housing functions as orchestrators of households' residential mobility in the context of Swiss rental housing. We propose a theoretical multi-step model and use survey data from 878 Swiss tenants to inspect the model's linkages. From the statistical analysis, we firstly observe that tenants' residential satisfaction is more likely to increase when the gap between ideal housing functions and those actually fulfilled by the current dwelling decreases. Secondly, results show that the effectiveness of an event (e.g. a job opportunity) in triggering the move is significantly related to both residential satisfaction and the functions the dwelling fulfils prior to the trigger. Thirdly, findings show that these trigger events can be grouped into three types: radical change, problem-solving and opportunity. With a medium effect size, a radical change was found to bring about the strongest change in housing functions between past and current dwellings. Lastly, in line with the hypothesis that residential preferences vary over the life course, socio-demographic characteristics and tenancy types are found to be significant explanatory variables for households' ideal housing functions. By disentangling the complexity of the housing system, the proposed multi-step model can be used to integrate households' preferences with supply-side constraints in agent-based model simulations, thereby contributing to fostering the provision of quality housing, i.e. dwellings able to meet the needs of current and future occupants.

Activities, Housing Situation and Other Factors Influencing Psychological Strain Experienced During the First COVID-19 Lockdown in Switzerland.

Background: The Coronavirus disease 2019 (COVID-19) crisis and the corresponding first nationwide lockdown from mid-March to 10 May 2020 engendered considerable psychological strain among people in Switzerland. This study analyzes determinants of changes in subjective levels of psychological strain experienced during the lockdown. Methods: An online survey conducted as part of a larger mixed methods study examined the material and emotional aspects of individual reactions to the lockdown from a socio-ecological perspective. Participants (N = 5932) were asked about their personal and employment status, housing features, changes in various activities (e.g., physical activity, watching TV, social media use) and aspects of mental distress and well-being. Results: A substantial share of participants reported to feel depressed (33%) and anxious (43%) more often during the COVID-19 lockdown than before, whereas significantly (p < 0.001) less persons reported a decrease of these negative feelings (depressed 17%; anxious 14%). Women, single people, students and people who lost their jobs or were temporally unemployed due to the lockdown experienced a particularly strong increase of subjective psychological strain. Important residential factors reducing subjective psychological strain were the general comfort of the housing situation and having a private garden or multiple types of outdoor space. Considering leisure activities, the strongest positive psychological effect resulted from increased physical activities, followed by reading and cooking. However, 45% of the participants reported a decreased frequency of physical activity during the lockdown compared to before, whereas significantly less persons (26%) reported a corresponding increase (p < 0.001). Conclusion: Consistent with other studies, the results indicate a substantial reduction of subjective psychological well-being of the population during the first COVID-19 lockdown in Switzerland. The psychological burdens which the participants experienced differ depending on personal characteristics and situational factors. Negative psychological and economic consequences and gender inequalities should accordingly be carefully considered and actively prevented when designing COVID-19 measures. Supportive economic and social, cognitive and behavioral psychological interventions need to be designed and implemented to maintain the well-being of residents during lockdown.

PCR-assisted impedimetric biosensor for colibactin-encoding pks genomic island detection in E. coli samples.

A fast PCR-assisted impedimetric biosensor was developed for the selective detection of the clbN gene from the polyketide synthase (pks) genomic island in real Escherichia coli samples. This genomic island is responsible for the production of colibactin, a harmful genotoxin that has been associated with colorectal cancer. The experimental protocol consisted of immobilizing the designated forward primer onto an Au electrode surface to create the sensing probe, followed by PCR temperature cycling in blank, positive, and negative DNA controls. Target DNA identification was possible by monitoring changes in the system's charge transfer resistance values (Rct) before and after PCR treatment through electrochemical impedance spectroscopy (EIS) analysis. Custom-made, flexible gold electrodes were fabricated using chemical etching optical lithography. A PCR cycle study determined the optimum conditions to be at 6 cycles providing fast results while maintaining a good sensitivity. EIS data for the DNA recognition process demonstrated the successful distinction between target interaction resulting in an increase in resistance to charge transfer (Rct) percentage change of 176% for the positive DNA control vs. 21% and 20% for the negative and non-DNA-containing controls, respectively. Results showed effective fabrication of a fast, PCR-based electrochemical biosensor for the detection of pks genomic island with a calculated limit of detection of 17 ng/µL.

Factor XIII Activity Might Already Be Impaired before Veno-Venous ECMO in ARDS Patients: A Prospective, Observational Single-Center Cohort Study.

Direct complications in patients receiving extracorporeal (veno-venous) membrane oxygenation (vvECMO) are mainly either due to bleeding or thromboembolism. We aimed to evaluate the course of routine coagulation parameters and the activity of different coagulation factors-with special focus on factor XIII (F XIII)-before, during and after vvECMO in acute respiratory distress syndrome (ARDS) patients. The activity of coagulation factors and rotational thrombelastometry were analyzed in 20 ECMO patients before (T-1) and 6 h (T0), one (T1), three (T3) and seven days (T7) after the implantation, as well as one and three days after the termination of ECMO. F XIII activity was already severely decreased to 37% (30/49) before ECMO. F XIII activity was the only coagulation factor continuously declining during vvECMO, being significantly decreased at T3 (31% (26/45) vs. 24% (18/42), p = 0.0079) and T7 (31% (26/45) vs. 23% (17/37), p = 0.0037) compared to T0. Three days after termination of vvECMO, platelet count and fibrinogen nearly doubled and factors II, V, XI and XIII showed spontaneous significant increases. Severe ARDS patients showed a considerably diminished factor XIII activity before vvECMO initiation and its activity continuously declined later on. Thus, incorporation of F XIII monitoring into the regular hemostaseologic routine during vvECMO therapy seems advisable. Due to the potential development of a hypercoagulatory state after the termination of vvECMO, tight hemostasiologic monitoring should persist in the initial phase after ECMO termination.

Leveraging insurance customer data to characterize socioeconomic indicators of Swiss municipalities.

The availability of reliable socioeconomic data is critical for the design of urban policies and the implementation of location-based services; however, often, their temporal and geographical coverage remain scarce. We explore the potential for insurance customers data to predict socioeconomic indicators of Swiss municipalities. First, we define a features space by aggregating at city-level individual customer data along several behavioral and user profile dimensions. Second, we collect official statistics shared by the Swiss authorities on a wide spectrum of categories: Population, Transportation, Work, Space and Territory, Housing, and Economy. Third, we adopt two spatial regression models exploring both global and local geographical dependencies to investigate their predictability. Results show consistently a correlation between insurance customer characteristics and official socioeconomic indexes. Performance fluctuates depending on the category, with values of R2 > 0.6 for several target variables using a 5-fold cross validation. As a case study, we focus on predicting the percentage of the population using public transportation and we discuss the implications on a regional scope. We believe that this methodology can support official statistical offices and it could open up new opportunities for the characterization of socioeconomic traits at highly-granular spatial and temporal scales.

Histopathological findings in the uteri and ovaries of clinically healthy cats presented for routine spaying.

OBJECTIVES: The aim of this study was to assess the histopathological findings in the uteri and ovaries from clinically healthy queens presented for elective spaying. METHODS: Ovaries and distal uterine horns or complete uteri from 106 female cats were evaluated for pathological alterations. RESULTS: Pathological alterations of the uterus and/or ovaries were evident in 29 cats; of these, corpora lutea were present on the ovaries of 15 cats. Ovarian cysts were found in 15 cats and were classified as cysts of the Wolffian or Muellerian ducts (n = 4), follicular cysts (n = 4), luteal cysts (n = 1), cystic rete ovarii (n = 2), combinations of different cysts (n = 2) and non-classifiable cysts (n = 2). In 21/106 cats, cystic endometrial hyperplasia (CEH) was present. The incidence of CEH increased with the age of the cat. Six cats had purulent endometritis with or without distension of the uterine lumen. Hyperplastic lesions of the endometrium were detected in two cats. In one cat, a uterine horn malformation with duplication of one uterine horn lumen was diagnosed. CONCLUSIONS AND RELEVANCE: Whereas the majority of ovarian cysts and slight-to-moderate CEH are unlikely to interfere with an animal's wellbeing, endometritis must be considered a serious health problem that requires veterinary attention.

Ovary cold storage and shipment affect oocyte yield and cleavage rate of cat immature vitrified oocytes.

In feline species, cooled transport of ovaries can be employed without detrimental effects to retrieve immature oocytes intended for in vitro embryo production purposes. Indeed, this is the most common way to collect gametes from gonads of wild, valuable animals after they die or are castrated far from specialized laboratories. However, fresh retrieved gametes are generally used, and their cryosensitivity is not known. This study employed ovariectomy-derived domestic cat gonads as a model for wild felids, and aimed to compare the yield and developmental competence of Cryotop-vitrified oocytes (VOs) collected and cryopreserved right after ovary excision (In loco-VOs) or after 24 h cooled transport of ovaries (Shipped-VOs). The number of collected oocytes was higher in In loco-VOs than in Shipped-VOs (mean ± SD: 8 ± 3.36 vs 5.6 ± 3.1, p = 0.05). In vitro embryo production resulted in similar maturation (35% for both vitrified groups, p = 1) and fertilization rates (In loco-VOs: 29.1%; Shipped-VOs: 22.2%; p = 0.295), but showed a difference in cleavage (In loco-VOs: 25.6%; Shipped-VOs: 14.5%; p = 0.0495). No differences were found in further embryo development. Taken together, results suggested that delayed oocyte vitrification after cooled transport of organs was feasible and allowed embryo development. However, the number of collected oocytes and the cleavage rate of matured oocytes were higher when oocyte vitrification was performed without delay after ovary excision, and this should be considered in gamete conservation programs for endangered felids.