Valuable female cattle are continuously subject to follicular puncture (ovum pick-up - OPU). This technique is commonly used for in-vitro embryo production, but may result in ovarian lesion. Mesenchymal stem cells (MSC) ameliorate the function of injured tissues, but their use to treat ovarian lesions in cattle has not been established. We investigated whether a local injection of MSC would reduce the negative effects of repeated OPU under acute and chronic scenarios in bovines. First, we performed four OPU sessions and injected 2.5 × 10⁶ MSCs immediately after the 4th OPU procedure (n = 5). The treated organs (right ovary) were compared to their saline-treated counterparts (left), and presented superior production of oocytes and embryos in the three following OPU sessions (P < 0.05). Then, cows with progressive fertility loss went through three OPU sessions. Animals received MSC, saline, or MSC + FSH in both ovaries after the first OPU. In the two following OPU sessions, the MSC and MSC + FSH - treated groups failed to present any significant alteration in the number of oocytes and embryos compared to saline-treated animals. Thus, MSC have beneficial effects on the fertility of OPU-lesioned cows, but not in cows with cystic ovarian disease and chronic ovarian lesions.

BACKGROUND Adenocarcinomas can be of several types, and MCF-7 is an adenocarcinoma of human breast cell line useful as preclinical model to screen therapeutic agents such as ultra-diluted Viscum album, an European plant whose extract is commonly used in cancer therapy. AIMS MCF-7 and mesenchymal stem cells were used to evaluate the in vitro cytotoxicity of homoeopathic Viscum album 1x10-3 (VAD3). METHODS cells were cultured for 24 hours in controlled environment (37.5oC and 5% CO2) in 96-well plates. After this time, VAD3 was added to the culture medium in concentrations varying from 10 to 100 ?L/mL for MTT assay (evaluation of viability of cells). A control group was maintained with culture medium only. After 48 hours, the procedures of analysis of cells viability were performed. RESULTS MTT assay showed that the concentrations of 42 ?L/mL and 62 ?L/mL were able to reduce cell viability to 50% in MCF-7 and mesenchymal stem cells, respectively, which means that half of the cells cultured were dead after 48 hours in contact with VAD3. CONCLUSION Viscum album presented higher cytotoxic action on human breast cancer cell line culture than on mesenchymal stem cells. This medicine is extensively used against cancer, and the use of the homoeopathic form of it brings new possibilities as no or fewer adverse effects would be present.

This study aimed to evaluate the safety of allogeneic adipose-derived mesenchymal stromal cell (aASC) treatment in dogs with chronic kidney disease (CKD) at the time of infusions and during the 120-day follow-up after the last infusion. Five dogs with CKD received three intravenous infusions of approximately 1×106?10% of aASCs per kilogram of body weight at 21-day intervals. Clinical and laboratory evaluations were performed at the time of each treatment and at 30 and 120 days after the last infusion. Adverse effects of the treatment were assessed using clinical observations, laboratory analyses, and owners’ answers about their dog’s behavior after infusions and during follow-up. The investigated animals did not present any adverse effects immediately after infusion or during the follow-up after the last infusion according to clinical and laboratory observations, as well as the dog owner’s descriptions. One treated animal showed a reduction in creatinine, from 3.5mg/dL to 2.4mg/dL from day 0 to day 153, gained 100g of body weight, and improved disposition. The study results demonstrate that aASC therapy is safe for dogs with CKD; however, further studies are needed to investigate these promising results.

The advances in regenerative medicine are very important for the development of medicine and the discovery of stem cells has shown a greater capacity to raise the level of therapeutic quality while their use becomes more accessible, especially in their mesenchymal form. In veterinary medicine, it is not different. The use of those cells, as well as recent advances related to the use of their extracellular vesicles, demonstrates a great opportunity to enhance therapeutic methods and ensure more life quality for patients, which can be in clinical or surgical treatments. Knowing the advances in these modalities and the growing clinical and surgery research and demands for innovations in orthopedic and neurology medicines, this paper aimed to review the literature about the methodologies of use and applications such as the pathways of action and the advances that were postulated for microvesicles and exosomes derived from mesenchymal stem cells in veterinary medicine, especially for musculoskeletal disorders and related injuries.

Despite the significance of adipose-derived stem cells in complementary and regenerative medicine, studies regarding clinical safety for allogeneic intravenous administration in healthy dogs remain insufficiently conducted. This study aimed to assess cellular viability through laboratory and microbiological evidence, along with genetic markers, in addition to evaluating clinic-laboratory safety for allogeneic intravenous administration of adipose-derived stem cells in healthy dogs. To achieve this, two cell batches and eight dogs were included in the study. The results revealed positive genetic markers for CD29, CD44, CD105, SOX2, and OCT3.4. Both batches exhibited positive cell differentiation into adipocytes, chondrocytes, and osteoblasts. Microbiological evidence showed negative results, and cell viability after thawing indicated 92% and 88.5% viable cells after 30 minutes, and 86% and 83.5% after 24 hours, respectively, for both batches. Hematological cell counts and serum biochemical enzyme levels, before and after intravenous treatment, did not exhibit statistical differences between the time points (p>0.05). Median values remained within the reference range for the species during and after 30 days of treatment. Based on the cellular viability results, observed patterns, and the absence of hematological side effects, it can be concluded that intravenous therapy with allogeneic adipose-derived stem cells is clinically safe.

Cumulative tissue damage and chronic inflammation associated with frequent ovum pickup (OPU) may lead to a progressive reduction in the number and quality of the oocytes recovered, particularly in donors with a high antral follicle count. The aim of this study was to evaluate the effect of an intraovarian treatment with mesenchymal stem cells (MSC) on oocyte yield, quality, and development potential during in vitro embryo production (IVEP) in cattle donors undergoing repeated OPU. Mesenchymal stem cells were previously isolated from adipose tissue, cultured in Dulbecco's modified Eagle medium until reaching 80% confluence, isolated with trypsin, and frozen in liquid N2 until use. Characterisation of MSC was carried out according to the guidelines of the International Society for Cellular Therapy. Nelore (Bos indicus) cows (n = 5) were used in this study, with the ovaries as replicates. The cows underwent eight OPU sessions at 15-day intervals, and the oocytes recovered were graded and used for IVEP with the semen of a single sire and batch under similar in vitro culture conditions. To ensure a high inflammatory response, immediately after the fourth OPU session all ovaries received 30 additional punctures, performed with a 16-gauge Jelco needle. Six hours later, the left ovary of each cow was injected with 500 µL of Dulbecco's modified phosphate buffered saline (control ovary) and the right ovary received 500 µL of Dulbecco's modified phosphate buffered saline with 2.5 × 106 allogenic MSC (treated ovary). Oocyte yield and embryo production before and after treatment were recorded for each ovary and donor. Grade I blastocysts produced from control and treated ovaries were used for gene expression evaluation. Data was analysed using the repeated-measures procedure of SAS (SAS Institute Inc.) to account for the effects of treatment, time, and interactions. There was no difference (P > 0.05) in any endpoint before treatment (sessions 1-4) between the right and left ovaries. Thus, differences between ovaries observed in OPU sessions 5-8 were assumed to be due to the treatment. After the injection of MSC, more total and viable oocytes were collected from the right ovaries compared with the left ovaries (15.3 ± 2.2 vs. 8.7 ± 1.2 (P < 0.02) and 13.6 ± 2.1 vs. 7.1 ± 1.0 (P < 0.01), respectively), resulting in more embryos produced in vitro (7.6 ± 1.2 vs. 3.6 ± 0.6, respectively; P < 0.01) as well as more initial and expanded blastocysts (1.4 ± 0.3 vs. 0.4 ± 0.1 and 4.4 ± 0.9 vs. 2.1 ± 0.4, respectively; P < 0.04). The proportion of viable oocytes recovered from the right ovary after treatment was greater than that from the left ovary (89.1% vs. 81.5%; P < 0.05). However, blastocyst rates did not differ between ovaries before or after treatment (50.4% vs. 55.5%: P > 0.05). In the blastocysts produced from treated ovaries, SLC2A3 was overexpressed (P < 0.04), whereas there was no difference for the expression of KRT8, PLAC8, SLC2A1, CASP3, PRDX3, or SOD2 (P > 0.05), suggesting potential differences in glucose uptake and metabolism. In conclusion, intraovarian treatment with MSC improved oocyte yield and quality and may be an alternative to increase IVEP from donors under intensive OPU schedules. This research was supported by CNPq, CAPES, and Fazenda Grupo Esplanada.