The invention relates to devices, compositions, and treatment of infertility inter alia by generating not only functional ovarian tissue but also functional oocytes from induced pluripotent stem cells. Treatments for hormone replacement are also described. In one aspect, the invention features a container including (i) a substrate; (ii) human iPSCs capable of differentiating into functional ovarian tissue; and (ii) a culture media.

Artificial ovaries comprising porous three-dimensional scaffolds are provided. Also provided are ink compositions and methods for printing the scaffolds. The artificial ovaries have spatial arrangements and cellular compositions that allow them to mimic native ovarian tissue. As such, they can be cultured or transplanted to support female endocrine function and/or the development of oocytes and/or eggs.

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.

The present invention relates to improved assisted reproductive technology using highly purified menotropin (HP-hMG) to stimulate follicle development in controlled ovarian stimulation, particularly in women at risk of a high ovarian response to controlled ovarian stimulation.

Disclosed herein are methods and compositions for treating or preventing ovarian failure using fibroblasts or cells derived from fibroblasts. In some embodiments, ovarian failure is pathological, the result of an intervention, or the result of aging. In some embodiments, regenerative fibroblast cells are administered locally into the ovary or pen-ovary areas or systemically. In some embodiments, regenerative cells act to suppress fibrosis of the ovaries, inhibit inflammation, stimulate maturation of immature ovarian progenitor cells, or directly differentiate into oocytes. In some embodiments, regenerative fibroblasts produce factors that inhibit apoptosis of oocytes and/or oocyte progenitors.

The present invention relates to a composition and method for assisted reproductive technology in mammals. In particular, the present invention provides compositions and methods for in vivo maturation of an immature cumulus oocyte complex (COC), thereby enhancing the embryology outcome.

A urine-derived mesenchymal stem cell mitochondria as well as a transplantation method and use thereof. The urine-derived mesenchymal stem cell mitochondrion is extracted from urine to be used for improving the quality of oocytes. The transplantation method comprises: jointly injecting sperms and the urine-derived mesenchymal stem cell mitochondria into mature oocytes for blastaea culture during the intracytoplasmic sperm microinjection. The present disclosure has the beneficial effects: during the traditional ICSI in combination with the transplantation of the urine-derived mesenchymal stem cell mitochondria, the fertilization rate of human in-vitro fertilization and the quality of embryos are significantly improved; the urine-derived mesenchymal stem cell mitochondria of the present disclosure can be used for in-vitro fertilization of low-prognosis patients with infertility with a good treatment effect; the problem of an autologous mitochondrion source in the prior art is solved without involving the introduction of a third-party genetic material and ethical issues.

A device for performing micro-operations on a vesicular target, comprising an injection pipette (27); a barrel assembly (9); and an outer assembly (1). The injection pipette, barrel assembly, and outer assembly being designed and situated in relation to each other such that an axial vacuum passage is created between the injection pipette and the barrel assembly, and a radial vacuum passage is created between the barrel assembly and outer assembly, and such vacuum passages are isolated from each other and from atmospheric pressure (FIG. 8). The device also comprises a means of advancing and withdrawing the distal end of the barrel assembly in relation to the distal end of the outer assembly so as to create a holding well (31) and a means of advancing the pipette into, and withdrawing the pipette from, a vesicular object.

The present invention relates to a method of producing a non-human, mammalian oocyte carrying a modified target sequence in its genome, the method comprising the steps of introducing into a non-human, mammalian oocyte: (a) a clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated protein 9 (Cas9 protein) or a nucleic acid molecule encoding said Cas9 protein; and (b-i) a target sequence specific CRISPR RNA (crRNA) and a trans-activating crRNA (tracr RNA) or a nucleic acid molecule encoding said RNAs; or (b-ii) a chimaeric RNA sequence comprising a target sequence specific crRNA and tracrRNA or a nucleic acid molecule encoding said RNA; wherein the Cas9 protein introduced in (a) and the RNA sequence(s) introduced in (b-i) or (b-ii) form a protein/RNA complex that specifically binds to the target sequence and introduces a single or double strand break within the target sequence. The present invention further relates to the method of the invention, wherein the target sequence is modified by homologous recombination with a donor nucleic acid sequence further comprising the step: (c) introducing a nucleic acid molecule into the cell, wherein the nucleic acid molecule comprises the donor nucleic acid sequence and regions homologous to the target sequence. The present invention also relates to a method of producing a non-human mammal carrying a modified target sequence in its genome.

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.