Haploid human embryonic stem cells and cell lines, haploid multipotent human cells, and haploid differentiated human cells are provided. In addition, methods of making and using the haploid human cells are provided.

Use invention provides a method for producing polypeptide protein products and nucleic acid products having reduced levels of antigenicity in an animal being treated with a biologic product. Somatic cells are isolated from an animal, transformed into pluripotent stem cells, transfected with a nucleic acid(s) of interest, and re-differentiated towards somatic cells known to be high level producers of the desired nucleic acid product. The invention can be used to derive a general cell line to treat populations, racial specific cell lines to treat ethnic groups, or patient specific cell lines to treat individuals. Additionally, the invention provides a method to allow induced pluripotent stem cells to be re-differentiated towards their somatic cell of origin so that the cells can be used to therapeutically treat an animal without resulting teratoma formation.

The present invention relates to a method for vitrification and thawing of oocytes of animals for somatic cell cloning. More specifically, the present disclosure relates to a method for vitrification and thawing of canine oocytes, and to thus produced frozen-thawed oocytes. In a conventional approach of the vitrification-frozen oocyte production for the dog, an estrous cycle may not coincide with an experimental schedule. However, the method for vitrification and thawing of the canine oocyte according to the present disclosure and the resulting frozen-thawed oocyte allows an experimental schedule to coincide with the estrous cycle, resulting in high nuclear transfer and fertilization effects.

Methods are provided for in vitro maturation (IVM) of bovine oocytes that include steps of (a) pre-culturing bovine germinal vesicle (GV) oocytes in the presence of C-type natriuretic peptide (CNP) and (b) subsequently culturing the oocytes of (a) for an extended duration in medium containing follicle stimulating hormone (FSH), luteinizing hormone (LH), 17-estradiol (E2), epidermal growth factor (EGF), and fetal bovine serum (FBS).

Provided are a method for preparing a mammalian ovum or embryo in which zona pellucida has been thinned or eliminated, and a method for fertilization using the mammalian ovum prepared by the aforementioned method. The resulting mammalian ovum or embryo is capable of realizing an improved fertilization rate and development rate when used for in vitro fertilization, transplantation of a fertilized ovum, or for preparation of an embryo in the early stages of development used in the production of a genetically modified animal.

The present invention generally relates to non-linear imaging systems and methods for use in various assisted reproductive technologies and other applications. In one aspect, the present invention is generally directed to two-photon and other non-linear microscope techniques that can be used to determine the status or maturity state of eggs or egg follicles, for example by determining the presence of polar bodies within the egg follicles. In some embodiments, endogenous metabolites, such as NADH or FAD, can be determined, and in some cases without externally altering those metabolite concentrations or without adding other compounds. Other embodiments of the invention are also directed to computer programs or systems that can be used to determine the status or maturity state of eggs or egg follicles, kits in such systems, or the like.

The present invention relates to compositions and methods for the handling of processed sperm including samples that are freshly collected, those transported as fresh samples, samples that are frozen and thawed, those sorted into one or more subpopulations, and those that are otherwise processed or handled that impose trauma on the cell. Trauma can reduce the motility, fertility, viability and overall integrity of the sperm and reduce the ability to fertilize, produce an embryo and a healthy offspring. The present invention relates to novel compounds that can be added to the sperm cell sample to reduce the traumatic effects of physical stress during mild as well as extensive sperm cell processing, methods of using the compounds in standard sperm processing procedures, the end products made from these methods including sperm and embryos, as well as methods of using those end products in assisted reproductive biology techniques in animals.

Compositions and methods comprising bioenergetic agents for restoring the quality of aged oocytes, enhancing oogonial stem cells or improving derivatives thereof (e.g., cytoplasm or isolated mitochondria) for use in fertility-enhancing procedures, are described.

An object of the present invention is to provide an agent for enhancing fertilization function of a mammal sperm, which comprises a low molecular compound which can be produced relatively easily and inexpensively as an active ingredient, and a method for enhancing fertilization function of a mammal sperm and a method for preparing a mammal fertilized egg, which use a low molecular compound which can be produced relatively easily and inexpensively. An agent comprising one or more compounds selected from the group consisting of compounds of the following formula (I.sub.0), formula (II), and formula (III), and physiologically acceptable salts thereof when R.sup.3 is OH is used as an agent for enhancing fertilization function of a mammal sperm. ##STR00001##

The present invention provides novel methods for improving the efficiency of artificial activation of unfertilized mammalian oocytes by reducing the intracellular concentration of Zn.sup.2+ in the oocyte. The methods of the invention may additionally comprise a preceding step of increasing the intracellular concentration of Ca.sup.2+ in the oocyte prior to reduction of the intracellular Zn.sup.2+ concentration. The invention further provides unfertilized oocytes activated by the disclosed methods and viable mammalian animals produced from unfertilized oocytes activated by the disclosed methods.