Valuable female cattle are continuously subject to follicular puncture (ovum pickup -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 × 10 6 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 OPUlesioned cows, but not in cows with cystic ovarian disease and chronic ovarian lesions.

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A doença renal crônica (DRC), enfermidade caracterizada pela inflamação túbulo-intersticial, atrofia tubular e fi-brose intersticial com perda progressiva da função renal, tem se tornado uma afecção relativamente comum em cães e gatos. Devido ao fato de o transplante renal ainda não ser uma opção viável na medicina veterinária, surgi-ram estudos para avaliar a possibilidade de tratamento de DRC com células-tronco mesenquimais (CTMs), em ra-zão dos efeitos anti-inflamatórios, anti-oxidantes, anti-fibróticos e imunomoduladores que essas células possuem. Onze pacientes portadores de DRC (6 cães e 5 gatos) foram avaliados e submetidos à terapia com CTMs halógenas. Todos os 5 gatos e 4 dos 6 cães tiveram diminuição nas taxas de creatinina sérica após o transplante de células, indicando que houve diminuição significativa para a creatinina em cães (p=0.0238). Houve, também, considerável diminuição na frequência de vômitos (p<0.05) e aumento no apetite (p<0.05) dos animais que receberam a terapia com células, de acordo com a percepção dos tutores. Esses resultados sugerem que a terapia com CTMs pode ser avaliada como uma alternativa para os animais portadores de DRC. Novos estudos devem ser realizados, com acompanhamento por maior espaço de tempo afim de estabelecer a melhor concentração de células e o número ideal de aplicações para estabilização dos pacientes portadores de DRC.

In vitro embryo production by handmade cloning (HMC) usually requires individual embryo culture, because zona-free embryos cannot be grouped in standard in vitro culture (IVC) protocols. The aim of this study was to evaluate the developmental potential of bovine embryos produced by HMC (Ribeiro et al. 2009 Cloning Stem Cells 11, 377–386) after in vitro culture (IVC) in 3 microwell (WOW) systems. After in vitro maturation, oocytes were denuded and incubated in demecolcine (Ibáñez et al. 2003 Biol. Reprod. 68, 1249–1258), followed by zona pellucida removal, oocyte bisection, embryo reconstruction, electrofusion, and chemical activation. Cloned embryos were allocated to 1 of 3 IVC groups: cWOW: conventional microwells (250 μm, round; Vajta et al. 2000 Mol. Reprod. Dev. 55, 256–264); mWOW: modified microwells (130 μm, conical; Feltrin et al. 2006 Reprod. Fert. Dev. 18, 126); and WOW-PDMS: microwells in polydimethylsiloxane chips (170 μm, cylindrical with microchannels); IVF embryos were used as controls (Bertolini et al. 2004 Reproduction 128, 341–354). Cleavage (Day 2), blastocyst (Day 7), and pregnancy (Day 30) rates were analysed by the chi-square test, for P < 0.05. Results are shown in Table 1. Cleavage rates were similar between groups, but development to the blastocyst stage was higher in IVF controls than cloned embryo groups. Among cloned embryo groups, blastocyst rate was higher in the mWOW group than the conventional and the PMDS-based microchannels. Nevertheless, in vivo development to Day 30 of pregnancy was not different between cloned groups. Our results for in vitro embryo development indicated that the mWOW provided more suitable conditions for embryo development to the blastocyst stage when compared with cWOW or even WOW-PDMS. Among some possible reasons include the physical advantage of a smaller microwell that may better mimic the constraining effect of the zona pellucida on the developing embryo. That may also provide greater blastomere stability, favouring the aggregation state during the first rounds of cleavages, also aiding compaction and subsequent cavitation. The narrower microwell system appeared to have promoted better in vitro development than the conventional and the DMPS-based microwell systems, with no impact on subsequent in vivo development. However, the IVC in the WOW-PDMS system supported reasonable rates of development, in accordance with the current literature. Table 1.In vitro development of bovine IVF and cloned embryos produced after the in vitro culture in distinct IVC systems

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