Disclosed herein include methods and compositions for culture medias for in vitro culture of synthetic embryos from mammalian pluripotent stem cells and extra-embryonic stem cells. The methods and compositions described herein can generate synthetic embryos at different developmental stage reaching early organogenesis and beyond. Disclosed herein also include an embryo culturing system and methods of using same.

Xenograft egg models comprising a fertilized non-mammalian egg comprising an ablated immune system, a first plurality of mammalian cells and a second plurality of mammalian cells, wherein the second plurality comprises immune cells are provided. Methods of producing the xenograft egg model as well as using the xenograft egg model for screening are also provided.

A computer-implemented system and method for predicting female sex bovine offspring to result from a bovine embryo by processing video image data of the embryo. The method includes receiving image data derived from video of a target embryo taken at substantially real-time frame speed during an embryo observation period of time. The video contains recorded morphokinetic movement of the target embryo occurring during the embryo observation period of time. The movement is represented in the received image data and the received image data is processed using a model generated utilizing machine learning and correlated embryo outcome data.

The invention provides a method for culturing mammalian cells. The method provides greater control over cell o growth to achieve high product titer cell cultures.

Provided is a temporary treatment medium for inhibiting embryo developmental arrest due to manipulation of an embryo or the like. A temporary treatment medium according to the present invention reduces damage to an in-vitro culture due to manipulation thereof, said in-vitro culture containing one of pluripotent stem cells, reproductive cells, a fertilized egg, and an embryo or any combination thereof. For this purpose, the temporary treatment medium contains a cytoskeleton regulator and/or an apoptosis inhibitor. The cytoskeleton regulator and/or the apoptosis inhibitor is preferably an Rho kinase inhibitor. Specifically, a Rock inhibitor can be used as the Rho kinase inhibitor. The Rock inhibitor is, for example, Y-27632. The temporary treatment medium is used for treating an in-vitro culture for a specific period before and/or after manipulation involving damage thereto.

Provided are methods of deriving a mammalian livestock pluripotent stem cells line, by (a) ex-vivo culturing a mammalian livestock embryo of at least 7 days post-fertilization for a culturing period of at least 4 days and no more than until 21 days post-fertilization so at to obtain an embryo comprising epiblast cell and/or late stage pluripotent stem cell; (b) isolating from the embryo the epiblast cell and/or the late stage pluripotent stem cell, and (c) culturing the epiblast cell and/or the late-stage pluripotent stem cell under conditions suitable for expansion of undifferentiated mammalian livestock pluripotent stem cells to thereby obtain a population of mammalian livestock pluripotent stem cells. Also provided are isolated mammalian livestock pluripotent stem cells, and cells differentiated therefrom.

The method for automated cell positioning can include: sampling a video of a scene having a gamete, tracking the gamete, and positioning the gamete within a target region. The method can optionally include: determining attribute values for the gamete, selecting the gamete, reorienting the gamete, and/or manipulating the gamete, and/or any suitable steps.

The present invention provides methods and compostions to improve the efficiency of somatic cell nuclear transfer (SCNT). There is increasing evidence that the epigenetic state of donor nuclei has a significant impact on potential of nuclear transfer embryos to develop into blastocysts, from which pluripotent stem cells are derived. Strategic application of histone agents, capable of altering epigenetic state such as methylation, allows zygotic activation and robust blastocyst generation.

Provided is a method for preparing a diabetic dog model by means of gene editing technology, a diabetic dog model prepared therefrom, as well as cells and issues thereof. The method comprises the following steps: (1) obtaining a dog fertilized egg cell, which comprises a point mutation in GCK gene, for a diabetic dog model by means of gene editing; and (2) transplanting the dog fertilized egg cell into one fallopian tube of a female dog, in which both fallopian tubes have been flushed, to prepare a diabetic dog model comprising a point mutation in GCK gene.

Embodiments of the present invention provide plant-derived extracts as a replacement for traditional cryoprotectants used to freeze tissue and cells providing a method to decrease post-thaw damage as created by the cryoprotectant. For example, extracts from the genus Hippophae or other plant sources or compositions may be used as a cryoprotectant or may even be used to replace at least some of a traditional cryoprotectant.