Male wistar han rats

Male Wistar Han rats and male Wistar Kyoto rats, at the age of 7 weeks and weighing approximately 180–210 g, were acquired from Charles River (Sulzfeld, Germany). Rats were housed in five animals per standard laboratory cage with free access to food and water and maintained on a 12-h light/dark cycle (lights on at 8 am) under conditions of constant temperature (22 ± 2 °C) and humidity (45 ± 5%). This study was approved by the II Ethical Committee at the Maj Institute of Pharmacology at the Polish Academy of Sciences, Kraków, Poland.

Male Wistar Han Rats (12-week-old), purchased from Charles River and weighing between 250 and 400 g, were housed in a specific pathogen-free animal facility on a 12 h light/12 h dark regimen and fed a commercial diet (pellets) and acidified drinking water ad libitum.
A total of 40 animals were randomized into 3 groups: Group 1—UCB-MNC-sEV 1 × 10¹⁰ particles/mL (1 × 10⁹ total particles) (14 animals); group 2—UCB-MNC-sEV 1 × 10¹¹ particles/mL (1 × 10¹⁰ total particles) (14 animals); group 3—vehicle (12 animals). Each group was randomly sub-divided into 2 groups depending on the duration of treatment: 6 or 12 weeks. UCB-MNC-sEV (100 μL at 1 × 10¹⁰ or 1 × 10¹¹ particles/mL) or control (vehicle) were intravenously injected into the tail vein, twice a week, over 6 or 12 weeks. The regimen of application was chosen based on the typical treatment schedule employed for the care of chronic wounds. During the experiment, the animals’ weight was monitored, and signals of fighting, dehydration, excessive barbering, or malocclusion were closely observed. After 6 or 12 weeks, the animals were euthanized and the organs and blood were collected, as described above.

Male Wistar Han rats (Charles River, Germany), weighing approximately 200–220 g, were housed as previously described by Wigner et al. (2020) [38 (link)]. All tests in the study were approved by the Bioethical Committee at the Institute of Pharmacology of the Polish Academy of Sciences in Krakow (Poland) and were conducted in compliance with the rules and principles of the 86/609/EEC Directive.

Male Wistar Han rats (Charles River, Lindau/Bodensee, Germany), weighing approximately 200–220 g, were individually housed under standard conditions, i.e., room temperature (22 °C) with twelve-hour cycles of day and night, with unlimited access to food and water. Each studied group consisted of six animals. All experimental procedures were carried out in accordance with the rules of the 86/609/EEC Directive and were approved by the Local Bioethics Commission of the Institute of Pharmacology, Polish Academy of Sciences in Krakow.

Male Wistar–Han rats 2 months of age (Charles River Laboratories, L’Arbresle, France) were used for all in vivo experiments. Those animals were housed in groups and kept under standard laboratory conditions, i.e., 22 ± 1 °C, 12 h light/dark cycle, 55% relative humidity, and ad libitum access to water and food. Rats from three independent cohorts were randomly divided into four groups (n = 14–16 per group were used for behavioral analysis, of which 6–8 were used for gene expression quantification, immunofluorescence, and morphologic studies. In detail, we used samples from three independent cohorts for a short-term analysis (immediately after stress exposure—tp1) and samples from two independent cohorts for a long-term analysis (4 weeks after stress exposure—tp2), for gene expression quantification, immunofluorescence, and morphologic purposes. All the procedures were conducted under the European Directive 2010/63/EU, and experiments were approved by the University of Minho Subcommittee of Ethics for the Life and Health Sciences (SECVS068/2017).

Male Wistar-Han rats (Charles River, Sulzfeld, Germany) weighting from 280 to 350 g were used in all performed experiments. Animals were initially acclimatized and housed (5 per cage) in environmentally controlled rooms under 12-h light/dark cycle (the light was switched on at 6 a.m.) at a temperature of 23 ± 1 °C and humidity of 55 ± 10%. Rats had free access to typical laboratory food and tap water (VRF 1, Special Diets Services, Witham, UK), enriched environment was not applied. The studies strictly conformed to European regulations for animal experimentation (EU Directive 2010/63/EU on the protection of animals used for scientific purposes). The experimental protocols were approved by the Local Ethics Commission for Experimentation on Animals (permit nos. 186 and 189/2017). This article does not contain any studies with human participants by any of the authors.

Male Wistar Han rats (Charles River, Saint-Germain-Nuelles, France) were housed under standard conditions of temperature (21–24 °C), humidity (40–60%) and a 12 h light/dark cycle with the light period starting at 07:00 a.m. Food and water were available ad libitum. Endotoxemic shock was induced in 28-days-old Wistar Han rats and compared to control rats (injection of NaCl 0.9%—CTRL) as previously described [14 (link)]. One hour after LPS injection, the rats were randomly assigned to no therapy (LPS) or fluidotherapy (NaCl 0.9%, 10 mL/kg—LPS + R) ± NButGT (NButGT, 10 mg/kg—NButGT) to increase O-GlcNAcylation levels (Figure 6).