The electrical pacemakers currently being used for the therapeutic approaches for treatment of “sick sinus syndrome” are not hormonally regulatable and entail risks through infections or premature battery discharge. These problems could be overcome by means of “biological cardiac pacemakers” obtained from pluripotent stem cells (PSCs). It has been shown that the controlled differentiation of stem cells with TBX, inductors of sinoatrial node cells, and an additional Myh6 promoter-specific antibiotic selection can give cardiomyocyte aggregates consisting to an extent of more than 80% of physiologically functional pacemaker cells. These induced sinoatrial bodies (“iSABs”) for the first time exhibited very high beat frequencies (300-400 bpm), similar to those in a murine heart, and were able to stably rhythmically stimulate heart muscle cells ex vivo. In the iSAB transcriptome decoded by means of RNA-seq, it was possible to assign almost all the genes to the ontologies of heart function/heart development and the structures of contractile cells. Overall, this is the first example of a high-purity functional sinoatrial tissue derived from stem cells, which means that a crucial step for future cell therapy and the testing of medicaments in vitro is being implemented.

Use of Cistanche deserticola polysaccharide (CDP) in promoting the proliferation and differentiation of female germline stem cells (FGSCs) is provided. Specifically, the addition of CDP in an in vitro cultivation system can promote the proliferation and differentiation of FGSCs, and especially can enhance the in vitro directed differentiation of FGSCs into oocytes, which provides a new research reference for studying the generation of oocytes in vivo and in vitro and also brings a new hope for research on physiological infertility.

Provided herein are methods and compositions relating, in part, to the generation and isolation of human lung progenitor cells from pluripotent stem cells.

The presently disclosed subject matter provides for in vitro methods of inducing differentiation of human stem cells into neural crest, cranial placode or non-neuro ectoderm precursors, and cells generated by such methods. The presently disclosed subject matter also provides for uses of such cells for treating neurodegenerative and pituitary disorders.

A method of providing a culture of oogonia stem cells comprising oogonia stem cells is provided. The method may further include culturing the oogonia stem cells with granulosa and theca cells to differentiate the oogonia stem cells into oocytes. In some embodiments, the culturing comprises including the oogonia stem cells in an in vitro follicle construct or a microcapsule comprising said granulosa and theca cells. Further described herein is a method of forming a bioengineered follicle construct capable of releasing a mature oocyte. An in vitro fertilization method using the mature oocyte is also provided.

The invention relates to a method for polar body injection, which comprises removing a polar body from a first egg cell and injecting the polar body into a second egg cell that is in a fertilizable state.

A problem of this invention it to provide a method for differentiate a primordial germ cell into a functional GV stage oocyte by in vitro culture. This invention relates to a method for differentiating a primordial germ cell into a functional GV stage oocyte by in vitro culture, comprising: (a) a step of producing a secondary follicle by culturing the primordial germ cell and supporting cells adjacent to the primordial germ cells under conditions that eliminate the effects of estrogen or a factor having a similar function to estrogen; (b) a step of partially dissociating cells between a granulosa cell layer and a thecal cell layer, wherein an oocyte, the granulosa cell layer, and the thecal cell layer constitute the produced secondary follicle; and (c) a step of differentiating the oocyte into a functional GV stage oocyte by culturing the oocyte, the granulosa cell layer, and the thecal cell layer that constitute the secondary follicle in a medium containing a high-molecular-weight compound.

Use of Cistanche deserticola polysaccharide (CDP) in promoting the proliferation and differentiation of female germline stem cells (FGSCs) is provided. Specifically, the addition of CDP in an in vitro cultivation system can promote the proliferation and differentiation of FGSCs, and especially can enhance the in vitro directed differentiation of FGSCs into oocytes, which provides a new research reference for studying the generation of oocytes in vivo and in vitro and also brings a new hope for research on physiological infertility.

The presently disclosed subject matter provides for in vitro methods of inducing differentiation of human stem cells into neural crest, cranial placode or non-neuro ectoderm precursors, and cells generated by such methods. The presently disclosed subject matter also provides for uses of such cells for treating neurodegenerative and pituitary disorders.

Provided a method for preparing a blastocyst. The method includes culturing a fertilized egg in a culture medium containing bubbles containing a reducing gas. The reducing gas may contain at least one selected from the group consisting of hydrogen, carbon monoxide, nitrogen monoxide, and hydrogen sulfide. The bubbles may have an average particle size of from 10 nm to 1,000 nm.