Provided herein are methods of producing β cells and precursors thereof utilizing a Wnt signaling inhibitor or PKC activator, or both. Also provided herein are in vitro cultures comprising said cells, methods of treating a subject with a disease characterized by high blood sugar levels over a prolonged period of time by administering said cells, and devices for encapsulating said cells.

This disclosure describes methods for organoid generation including, for example, for generation of a multi-tissue organoid. The multi-tissue organoid may include cartilage, bone, epithelium, and/or fibrous connective tissue. This disclosure further describes methods for isolating cells from the organoids and methods of using the organoids and cells of the organoids.

Provided herein are methods of producing β cells and precursors thereof utilizing a Wnt signaling inhibitor or PKC activator, or both. Also provided herein are in vitro cultures comprising said cells, methods of treating a subject with a disease characterized by high blood sugar levels over a prolonged period of time by administering said cells, and devices for encapsulating said cells.

The present invention provides methods to promote differentiation of pluripotent stem cells to pancreatic endoderm cells expressing PDX1, NKX6.1, and HB9. In particular, the methods encompass culturing Stage 4 to Stage 6 cells with a thyroid hormone (e.g. T3), an ALK5 inhibitor, or both.

The present invention describes in vitro methods for producing a cellular composition with in vivo bone forming potential.

An agent capable of promoting proliferation and differentiation of granulosa cells is disclosed which comprises a growth and differentiation factor-9 (GDF9) protein comprising a modified GDF9 polypeptide monomer which includes at least one amino acid substitution that enhances binding to and/or activation of activin-like kinase 4 and/or 5 receptor (ALK4/5). The agent is preferably provided in a mature dimeric form (eg comprising two monomers of the same modified GDF9 polypeptide monomer) and/or in a pro/mature complex form. The agent may be suitable for, inter alia, promoting oocyte maturation in vitro for use in assisted reproductive technologies.

The present invention provides methods to promote differentiation of pluripotent stem cells to pancreatic endoderm cells expressing PDX1, NKX6.1, and HB9. In particular, the methods encompass culturing Stage 4 to Stage 6 cells with a thyroid hormone (e.g. T3), an ALK5 inhibitor, or both.

A method of banking stem cells by harvesting stem cells from an individual and culturing the harvested stem cells to generate a P0 culture. The method includes storing a desired amount of P0 stem cells via cryopreservation and subculturing a portion of the P0 stem cells to generate a P1 culture. A desired amount of P1 stems cells can then be stored via cryopreservation and further generations can be stored as desired.

The present invention relates to the targeted engineering of specific cell populations. Motoneurons (MN) subtypes display differential vulnerabilities in diseases and in spinal injuries. Engineered MNs of specific rostro-caudal identity represent an important resource for cell therapy approaches. However, these strategies remain impeded by slow and inefficient targeted differentiations due to the imprecise control over cell fate specification in vitro. The inventors now used an embryoid body-based differentiation of hPSC and showed that the HOX clock expression can be controlled to generate subtypes of spinal MNs. Thus, the present invention relates to an in vitro or an ex vivo method for producing spinal neuronal subtypes comprising exposing axial progenitors to retinoic acid (RA), an agonist of Hedgehog signalling pathway, and optionally a FGFR agonist and/or an activator of the TGF pathway, wherein more and more caudal motor neurons identities are obtained by delayed exposure to RA and/or by exposure to RA in combination with the FGFR agonist and/or the activator of the TGF pathway.

Aspects of the present invention include methods and compositions related to the production and use of clonal lineages of embryonic progenitor cell lines derived from differentiating cultures of primordial stem cells, in particular, said methods and compositions relate to methods of differentiating cells in the presence of members of the BMP family of growth factors and the applications of said cell lines in the treatment of degenerative orthopedic diseases such as osteoarthritis.