Optimised CO2-containing medium for in vitro culture and transportation of mouse preimplantation embryos without CO2 incubator.

Conventional in vitro culture and manipulation of mouse embryos require a CO2 incubator, which not only increases the cost of performing experiments but also hampers the transport of embryos to the other laboratories. In this study, we established and tested a new CO2 incubator-free embryo culture system and transported embryos using this system. Using an Anaero pouch, which is a CO2 gas-generating agent, to increase the CO2 partial pressure of CZB medium to 4%-5%, 2-cell embryos were cultured to the blastocyst stage in a sealed tube without a CO2 incubator at 37°C. Further, the developmental rate to blastocyst and full-term development after embryo transfer were comparable with those of usual culture method using a CO2 incubator (blastocyst rate: 97% versus 95%, respectively; offspring rate: 30% versus 35%, respectively). Furthermore, using a thermal bottle, embryos were reliably cultured using this system for up to 2 days at room temperature, and live offspring were obtained from embryos transported in this simple and very low-cost manner without reducing the offspring rate (thermal bottle: 26.2% versus CO2 incubator: 34.3%). This study demonstrates that CO2 incubators are not essential for embryo culture and transportation and that this system provides a useful, low-cost alternative for mouse embryo culture and manipulation.

PrintrLab incubator: A portable and low-cost CO2 incubator based on an open-source 3D printer architecture.

Growth in open-source hardware designs combined with the decreasing cost of high-quality 3D printers have supported a resurgence of in-house custom lab equipment development. Herein, we describe a low-cost (< $400), open-source CO2 incubator. The system is comprised of a Raspberry Pi computer connected to a 3D printer controller board that has controls for a CO2 sensor, solenoid valve, heater, and thermistors. CO2 is supplied through the sublimation of dry ice stored inside a thermos to create a sustained 5% CO2 supply. The unit is controlled via G-Code commands sent by the Raspberry Pi to the controller board. In addition, we built a custom software application for remote control and used the open-source Grafana dashboard for remote monitoring. Our data show that we can maintain consistent CO2 and temperature levels for over three days without manual interruption. The results from our culture plates and real-time PCR indicate that our incubator performed equally well when compared to a much more expensive commercial CO2 incubator. We have also demonstrated that the antibiotic susceptibility assay can be performed in this low-cost CO2 incubator. Our work also indicates that the system can be connected to incubator chambers of various chamber volumes.

Comparação de métodos in vitro para a quantificação da digestibilidade da matéria seca e da fibra em detergente neutro de forragens e concentrados / Comparison of in vitro methods to quantify the dry matter and neutral detergent fiber digestibility of forages and concentrates

Objetivou-se comparar a digestibilidade in vitro da matéria seca (DIVMS) e da fibra em detergente neutro (DIVFDN) de forragens e concentrados obtidos por intermédio das incubadoras DaisyII e TE-150, combinadas com filter bags tipo F57 (Ankon®) ou tecido não-tecido (TNT, 100g/m²), em relação aos valores obtidos utilizando-se o método de Tilley e Terry (1963). Foram utilizadas 25 amostras de alimentos concentrados e 25 de forragens. Não houve diferenças entre filter bags (P>0,39) para a DIVMS de forragens. Maiores valores de DIVMS (P<0,01) foram verificados para a incubadora TE-150 em relação à DaisyII. As estimativas de DIVMS obtidas com incubadoras e filter bags foram superiores (P<0,01) àquelas obtidas com o método Tilley e Terry (1963). Observaram-se maiores valores de DIVFDN de forragens (P<0,01) com filter bags F57 em relação ao TNT e com a incubadora TE-150 em relação à DaisyII. Todos os valores de DIVFDN obtidos com incubadoras e filter bags foram superiores (P<0,01) aos obtidos com o método Tilley e Terry (1963). Todos os métodos apresentaram-se positiva e fortemente correlacionados (P<0,01), tanto para DIVMS como para DIVFDN. As repetibilidades e variâncias entre amostras para DIVMS e DIVFDN foram similares entre os métodos. Portanto, conclui-se que, em se tratando de avaliações comparativas entre alimentos, todos os métodos avaliados possuem capacidade similar de discriminação.(AU)

The objective was to compare in vitro digestibility of dry matter (IVDMD) and neutral detergent fiber (IVNDFD) of forages and concentrates obtained using the incubators DaisyII and TE-150, and filter bags F57 (Ankon®) or non-woven textile (NWT - 100g/m²) in comparison with the values obtained using the Tilley and Terry (1963) method. Twenty-five samples of concentrates and 25 samples of forages were used. There were no differences between filter bags (P>0.39) for IVDMD of forages. Higher IVDMD (P<0.01) were observed for TE-150 compared to DaisyII. Estimates of IVDMD obtained with incubators and filter bags were higher (P<0.01) than those obtained with the Tilley and Terry (1963) method. Higher IVNDFD (P<0.01) were obtained with filter bags F57 regarding to NWT and TE-150 when compared to DaisyII. All values of the IVNDFD obtained with incubators and filter bags were higher (P<0.01) than those obtained with the Tilley and Terry (1963) method. Both IVDMD and IVNDFD variables were greater using TE-150 incubator than using DaisyII (P<0.04). Estimates obtained using NWT filter bags were higher (P<0.01) than F57 filter bags. All methods were positively and strongly correlated (P<0.01) to each other for IVDMD and IVNDFD. The repeatability and variances among samples for IVDMD and IVNDFD were similar among methods. In conclusion, all methods were similarly able to discriminate samples.(AU)

Obstetric and perinatal outcomes of pregnancies conceived with embryos cultured in a time-lapse monitoring system.

OBJECTIVE: To compare obstetric and perinatal outcomes of singleton pregnancies resulting from embryos incubated in a time-lapse system (TLS) with those of embryos grown in standard IVF incubators (SI). DESIGN: Retrospective description of a cohort of patients who conceived during a randomized, controlled trial. SETTING: Private university-affiliated IVF center. PATIENT(S): Of 856 randomized patients, 378 gave birth to a live-born infant: 216 of the deliveries originated from embryos incubated in TLS, and 162 deliveries were from embryos cultured in SI. INTERVENTION(S): Embryo incubation and selection in TLS. MAIN OUTCOME MEASURE(S): Delivery and neonatal outcomes. RESULT(S): No significant differences were observed in the baseline characteristics of the study population. The delivery rate was 49.3% (TLS) vs. 40.0% (SI), and multiple deliveries were higher in the TLS group: 31.0% (67 of 216) vs. 24.7% (40 of 162) in the SI group. When singleton pregnancies were analyzed no differences were found between the two groups in the rate of obstetric problems with respect to weeks at delivery: 38.8 (95% confidence interval [CI] 38.4-39.1) (TLS) vs. 39.5 (95% CI 38.0-39.9) (SI); preterm births (<37 weeks): 10.7% (TLS) vs. 12.3% (SI); and very preterm births (<34 weeks): 2.9% (TLS) vs. 3.3% (SI). No statistical differences were found in neonatal outcomes such as birth weight: 3,163 g (95% CI 3,035-3,292 g) (TLS) vs. 3,074 (95% CI 2,913-3,236) (SI); low birth weight (<2,500 g): 12.8% (TLS) vs. 12.3% (SI); very low birth weight (<1,500 g): 2.0% (TLS) vs. 2.4% (SI); or height: 50.3 cm (95% CI 49.6-50.9 cm) (TLS) vs. 49.7 (95% CI 48.9-50.4 cm) (SI). No major malformations or perinatal mortality were found in either of the two groups. CONCLUSION(S): No detrimental effects were observed in obstetric and perinatal outcomes when a time-lapse incubator was used rather than a more widely used conventional incubator. As far as we know this is the first report from a randomized study of the neonatal outcomes of time-lapse monitoring. Our results suggest that this technology is an effective and safe alternative for embryo incubation, though trials of larger numbers of patients are required to further confirm our conclusions. CLINICAL TRIAL REGISTRATION NUMBER: NCT01549262.

Humid versus dry incubator: a prospective, randomized, controlled trial.

OBJECTIVE: To evaluate the efficacy of a dry versus humidified incubator on human embryo development ex vivo. DESIGN: Prospective, double-blind, randomized, controlled trial. SETTING: Private fertility centers. PATIENT(S): A total of 297 women undergoing in vitro fertilization randomized into two groups. INTERVENTION(S): From days 0 to day 5 or 6 of culture, intervention group embryos exposed to dry culture and control group embryos exposed to humidified culture. MAIN OUTCOME MEASURE(S): Subsequent ongoing pregnancy rate. RESULT(S): After transfer of embryos, there were statistically significantly lower rates of clinical and ongoing pregnancy in the dry culture arm than in the humidified culture arm (odds ratio [OR] 0.57; 95% confidence interval [CI], 0.36-0.91; versus OR 0.54; 95% CI, 0.34-0.85). On day 3 of culture, embryo quality and compaction were lower in the dry culture group (OR 0.38; 95% CI, 0.32-0.45) than in the group exposed to humidified culture (OR 0.23; 95% CI, 0.19-0.27). On day 5 of culture, embryos in dry culture had a lower rate of blastocyst formation (OR 0.39; 95% CI, 0.33-0.46), quality (OR 0.34; 95% CI, 0.29-0.40), and cryopreservation (OR 0.41; 95% CI, 0.35-0.48). CONCLUSION(S): In this study, human embryos cultivated ex vivo in a dry incubator had statistically significantly decreased implantation and clinical and ongoing pregnancy rates. Our findings indicate the need for larger multicenter, randomized, controlled trials. CLINICAL TRIAL REGISTRATION NUMBER: NCT01695096.

Níveis de ruído das incubadoras das unidades neonatais de um hospital universitário de Ribeirão Preto-SP

Este estudo insere-se em um projeto mais amplo dirigido à prevenção de ruídos nas unidades neonatais do hospital universitário, público, situado no município de Ribeirão Preto – SP. Está voltado para o ruído em incubadoras, apresentando como objetivos: a quantificação dos níveis de ruído contínuo e de impacto desses equipamentos nas unidades neonatais de cuidados intensivo e intermediário desse hospital e a identificação da sistemática de manutenção das mesmas, enfatizando os aspectos pertinentes ao controle de ruídos. O estudo populacional é do tipo experimento não-controlado. O parâmetro utilizado para o nív elm áxim o de ruído contínuo, permitido dentro da incubadora, foi de 60 dB/A NPS, de ac ordo c om a Norma Brasileira IEC 601-2-1997....