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Background: Conditioned medium is the medium obtained from certain cultured cells and contained secretome from the cells. The secretome, which can be in the form of growth factors, cytokines, exosomes, or other proteins secreted by the cells, can induce the differentiation of cells that still have pluripotent or multipotent properties., Objectives: This study examined the effects of conditioned medium derived from E17 rat brain cells on cells with pluripotent properties., Methods: The conditioned medium used in this study originated from E17 rat brain cells. The CM was used to induce the differentiation of primary colonies of mice blastocysts. Primary colonies were stained with alkaline phosphatase to analyze the pluripotency. The morphological changes in the colonies were examined, and the colonies were stained with GFAP and Neu-N markers on days two and seven after adding the conditioned medium., Results: The conditioned medium could differentiate the primary colony, beginning with the formation of embryoid-body-like structure; round GFAP positive cells were identified. Finally, neuron-like cells testing positive for Neu-N were observed on the seventh day after adding the conditioned medium., Conclusions: Conditioned medium from different species, in this case, E17 rat brain cells, induced and promoted the differentiation of the primary colony from mice blastocysts into neuron-like cells. The addition of CM mediated neurite growth in the differentiation process., Competing Interests: The authors declare no conflicts of interest., (© 2021 The Korean Society of Veterinary Science.)

In vitro fertilization has been regarded as a forefront solution in treating infertility for over four decades, yet its effectiveness has remained relatively low. This could be attributed to the lack of advancements for the method of observing and selecting the most viable embryos for implantation. The conventional morphological assessment of embryos exhibits inevitable drawbacks which include time- and effort-consuming, and imminent risks of bias associated with subjective assessments performed by individual embryologists. A combination of these disadvantages, undeterred by the introduction of the time-lapse incubator technology, has been considered as a prominent contributor to the less preferable success rate of IVF cycles. Nonetheless, a recent surge of AI-based solutions for tasks automation in IVF has been observed. An AI-powered assistant could improve the efficiency of performing certain tasks in addition to offering accurate algorithms that behave as baselines to minimize the subjectivity of the decision-making process. Through a comprehensive review, we have discovered multiple approaches of implementing deep learning technology, each with varying degrees of success, for constructing the automated systems in IVF which could evaluate and even annotate the developmental stages of an embryo., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

An investigation was conducted to determine the influence of two sperm selection modalities, IMSI and ICSI, on the morphokinetics, dynamic development and ploidy status of embryos derived from males with sub‐optimal sperm profiles during IVF program. A total of 209 PGTA‐tested top‐quality blastocysts (IMSI = 129, ICSI = 80) from 84 couples (IMSI = 51, ICSI = 33) were assessed retrospectively. This study found that both IMSI and ICSI yielded comparable embryo morphokinetics, except for the T7, TEB and CC3 parameters (p < 0.05). A significant lower incidence of multinucleation was observed in the IMSI group when compared to the ICSI group (48.8% vs. 71.3%, p = 0.002), while other parameters of embryo development such as direct cleavage, distorted cytoplasmic movement, reverse cleavage and vacuole(s) appearance did not differ (p > 0.05). No differences were noticed in the proportion of generating chromosomally euploid embryos (44.2% vs. 51.3%, p = 0.394, respectively, for IMSI and ICSI). The implementation of IMSI or ICSI in couples with sub‐optimal sperm profiles resulted in embryos with comparatively similar morphokinetics. Furthermore, the incidence of multinucleation at the two‐ to four‐cell stage was lower following the practice of IMSI, although the method did not improve the proportion of gaining euploid embryos. [ABSTRACT FROM AUTHOR]

Parthenogenesis is an artificial oocytes activation process without paternal contribution. Blastocyst, derived from parthenogenesis, is one of potential source for pluripotent stem cell propagation. Unfortunately, previous studies reported that parthenogenetic embryo did not achieve exhilarating blastocyst rate. One of the component that predicted inhibit parthenogenetic embryo development is phosphate. Therefore, we try to modify culture medium in order to overcome that problem. The aim of this research was to produce and analyze the characteristics of parthenogenetic blastocyst developed in phosphate-free medium. Mouse oocytes obtained from adult female DDY by superovulation. The activator was strontium chloride 10 mM and diploidization with cytochalasin B 5 μg/ml. Medium for activation and culture medium were modified rat 1 cell embryo medium (MR1ECM) which is phosphate free. The results showed that parthenotes that were cultured in phosphate free medium reached higher blastocyst rate compared to the other groups. The increase of phosphate in culture medium lead to impaired parthenogenetic embryos development. Further experiment was made to analyze the differences between fertilized and parthenogenetic embryo in this medium. The experiment showed that diploid parthenogenetic could achieve high blastocyst rate (30.9±1.3%). The quality of diploid parthenogenetic blastocyst, based on cells number, viability, and ICM ratio, was lower than fertilized blastocyst. [ABSTRACT FROM AUTHOR]

Vitrification is one of cryopreservation method for freezing cells without ice formation so that biological materials such as sperm, oocyte, or embryo, can be preserved and later can be used for a specific purpose including wildlife conservation efforts. Unfortunately, high concentration of cryoprotectan in vitrification process can cause osmotic stress and has high toxic levels that may affect embryo quality. The purpose of this research is to analyze the quality of post-vitrification embryos at morula and blastocyst stages based on morphometric variable, and Bax gene expression, futhermore potency of implantation of the post-vitrification embryo were also examined based on H19 and Igf2 gene expression. The results showed embryo viability postvitrification decreased 5.67% and 6.02% in morula and blastocyst. Development ability from morula to blastocyst post-vitrification was also decreased by 10.15%. Morphometry analysis of morula post-vitrification showed decreased values of zona pellucida tickness (ZPT), zona pellucida tickness variation (ZPTV) and blastomeres area, while perivetellin space (PVS) area was increased compared to fresh morula. Blastocyst post-vitrification had increased values of ZPT, ZPTV, and PVS, while in ICM and the blastocoel were also decreased compared to fresh blastocyst. The result of relative levels mRNA of Igf2, H19, and Bax gene show no significant difference gene expression between fresh blastocyst, blastocyst post-vitrification, and morula postvitrification group. [ABSTRACT FROM AUTHOR]

Background: Several approaches have been proposed to optimize the construction of an artificial intelligence-based model for assessing ploidy status. These encompass the investigation of algorithms, refining image segmentation techniques, and discerning essential patterns throughout embryonic development. The purpose of the current study was to evaluate the effectiveness of using U-NET architecture for embryo segmentation and time-lapse embryo image sequence extraction, three and ten hr before biopsy to improve model accuracy for prediction of embryonic ploidy status. Methods: A total of 1.020 time-lapse videos of blastocysts with known ploidy status were used to construct a convolutional neural network (CNN)-based model for ploidy detection. Sequential images of each blastocyst were extracted from the time-lapse videos over a period of three and ten hr prior to the biopsy, generating 31.642 and 99.324 blastocyst images, respectively. U-NET architecture was applied for blastocyst image segmentation before its implementation in CNN-based model development. Results: The accuracy of ploidy prediction model without applying the U-NET segmented sequential embryo images was 0.59 and 0.63 over a period of three and ten hr before biopsy, respectively. Improved model accuracy of 0.61 and 0.66 was achieved, respectively with the implementation of U-NET architecture for embryo segmentation on the current model. Extracting blastocyst images over a 10 hr period yields higher accuracy compared to a three-hr extraction period prior to biopsy. Conclusion: Combined implementation of U-NET architecture for blastocyst image segmentation and the sequential compilation of ten hr of time-lapse blastocyst images could yield a CNN-based model with improved accuracy in predicting ploidy status. [ABSTRACT FROM AUTHOR]