Methods for differentiating pluripotent stem cells to neuroectoderm in dynamic suspension culture using small molecule or protein inhibitors of TGFβ/Activin/Nodal signaling and BMP signaling are provided. Also provided are methot and protocols for differentiating pluripotent stem cells such as human embryonic stem cells first to neuroectoderm, then further to glial progenitor cells, and further to oligodendrocyte progenitor cells (OPCs), and compositions obtained thereby. The methods of the present disclosure reproducibly produce neuroectoderm progenitor cells by day 7 of the differentiation process, glial progenitor cells by day 21 of the differentiation process and OPCs by day 42 of the differentiation process.

Methods of producing hepatocyte lineage cells are provided. The method can include transfecting a cell with one or more expression vectors. For example, a cell can be transfected with a first expression vector containing a first gene that encodes CCAAT/enhancer binding protein alpha (CEBPA), a second expression vector containing a second gene that encodes CCAAT/enhancer binding protein beta (CEBPB), a third expression vector containing a third gene that encodes forkhead box A1 (FOXA1), and a fourth expression vector containing a fourth gene that encodes forkhead box A3 (FOXA3). The method can include culturing the transfected cell obtained in a growth environment. The transfected cell can be cultured in Williams' E medium, ReproFF (feeder-free media maintaining pluripotency) medium, or both. Transfected and/or hepatocyte lineage cells obtained by a method of the present invention are also provided.

The invention is directed to in vitro methods of inducing differentiation of anterior foregut endoderm and the enriched populations of anterior foregut endoderm produced by such methods. Such enriched populations are useful for studies of the molecular events that occur during differentiation and for generating cells for cell replacement therapy.

The present invention provides a method for producing a human cell aggregate containing a midbrain-hindbrain boundary neural progenitor tissue, including subjecting an aggregate of human pluripotent stem cells to suspension culturing in a serum-free medium containing insulin, and treating, in the suspension culturing, the aggregate of human pluripotent stem cells or a human cell aggregate derived therefrom with a ROCK inhibitor, a TGFβ signal inhibitor, and a first fibroblast growth factor. Furthermore, the human cell aggregate containing the midbrain-hindbrain boundary neural progenitor tissue is subjected to suspension culturing in a serum-free medium to induce formation of a neuroepithelial structure by neural progenitor in the neural progenitor tissue, whereby the human cell aggregate containing the cerebellar plate tissue can be obtained.

Provided herein are methods of manufacturing β cells in vitro. Also provided herein are methods of treating a disease in a subject comprising administering the β cells manufactured in vitro to the subject. Also provided herein are methods of differentiating stem cells into β cells.

Disclosed is a method for effectively preparing mesenchymal-like stem cells, the method including: preparing human pluripotent stem cells cultured to passage 70 or lower after establishment of cell lines; inducing differentiation of the human pluripotent stem cells to produce embryoid bodies and selecting cystic embryoid bodies therefrom; loading the cystic embryoid bodies on a cell-permeable three-dimensional (3D) culture unit to isolate mesenchymal-like stem cells therefrom; isolating only monolayer-shaped cell clusters from the mesenchymal-like stem cells passing through the cell-permeable 3D culture unit; and uniformizing sizes of the monolayer-shaped cell clusters, in which the mesenchymal-like stem cells have anti-inflammatory efficacy and immunosuppression. Further, disclosed are mesenchymal-like stem cells prepared by the preparation method, a transporter or a therapeutic composition including the mesenchymal-like stem cells, and a composition for prevention or treatment of diseases that includes an active ingredient or exosomes secreted from the mesenchymal-like stem cells.

Provided are a composition and a kit for differentiation of stem cells into neural progenitor cells (NPCs), each including a protein kinase C (PKC) inhibitor and a bone morphogenic protein (BMP) inhibitor, and a method using the same. Accordingly, stem cells may be efficiently differentiated into neural progenitor cells using a simple, low-cost composition.

Cells derived from postpartum tissue and methods for their isolation and induction to differentiate to cells of a chondrogenic or osteogenic phenotype are provided by the invention. The invention further provides cultures and compositions of the postpartum-derived cells and products related thereto. The postpartum-derived cells of the invention and products related thereto have a plethora of uses, including but not limited to research, diagnostic, and therapeutic applications, for example, in the treatment of bone and cartilage conditions.

Provided is an isolated population of human pluripotent stem cells comprising at least 50% human pluripotent stem cells characterized by an OCT4+/TRA1-60−/TRA1-81−/SSEA1+/SSEA4− expression signature, and novel methods of generating and maintaining same in a pluripotent, undifferentiated state a suspension culture devoid of cell clumps. Also provided are novel culture media, cell cultures and methods for culturing pluripotent stem cells in a suspension culture or a two-dimensional culture system while maintaining the cells in a proliferative, pluripotent and undifferentiated state. The novel culture media comprise interleukin 11 (IL11) and Ciliary Neurotrophic Factor (CNTF); bFGF at a concentration of at least 50 ng/ml and an IL6RIL6 chimera; or an animal contaminant-free serum replacement and an IL6RIL6 chimera. Also provided are methods for generating lineage-specific cells from the pluripotent stem cells.

The disclosure features methods and compositions for differentiating stem cells into hematopoietic stem and progenitor cells (HSPC) and/or Natural Killer (NK) cells. The methods and compositions described herein are used to differentiate stem or progenitor cells having at least one gene-edit that is maintained in the differentiated cell. Also provided are differentiated cells produced using the methods and compositions described herein for therapeutic applications.