The present invention relates to a stable method for introducing at least one inducible cassette into a cell, and permitting controllable transcription from within that inducible cassette. The method may be used for any cell type, from any eukaryotic organism, but has a particular application in the introduction of inducible cassettes into pluripotent stem cells, such as animal or human pluripotent stem cells (hPSCs). The inducible cassette is controllably inserted in such a way to ensure that the genetic material it contains is not silenced or subject to negative influences from the insertion site, and transcription of the genetic material is controlled.

Provided herein are methods of producing a distinct bilayer culture of retinal epithelial cells (RPE) with photoreceptor cells and/or photoreceptor precursor cells (PR/PRP). Further provided herein is a therapy comprising transplantation of the RPE and PR/PRP bilayer as well as methods for testing candidate drugs using the bilayer.

The present invention relates to 3D brain organoids, uses thereof, methods and culture medium for generating such organoids. An aspect of the invention provides brain organoids and methods of generating such organoids with bilaterally symmetric optic vesicles, containing both neuronal and non-neuronal cell types, and exhibiting functional circuitry. These organoids can be generated within short time intervals (e.g., 50 days) and therefore are useful for medical modelling and applications.

This disclosure relates to methods of producing induced pluripotent (iPS), multipotent, and/or lineage-committed stem cells from differentiated cells, maintaining iPS, multipotent, and/or lineage-committed cells in culture, and re-differentiating the iPS and multipotent stem cells into any desired lineage-committed cell type.

The invention provides a method for producing a cell aggregate containing a ciliary marginal zone-like structure by culturing a cell aggregate containing a retinal tissue in which Chx10 positive cells are present in a proportion of 20% or more of the tissue in a serum-free medium or serum-containing medium, each containing a substance acting on the Wnt signal pathway for only a period before the appearance of a RPE65 gene expressing cell, followed by culturing the “cell aggregate in which a RPE65 gene expressing cell does not appear” thus obtained in a serum-free medium or serum-containing medium, each not containing a substance acting on the Wnt signal pathway and so on.

An object of the present invention is to provide a human astrocyte cell population that is differentiated from astrocyte progenitor cells derived from human iPS cells, a manufacturing method for the human astrocyte cell population; and an evaluation method for a test substance using the human astrocyte cell population. According to the present invention, there is provided a human astrocyte cell population that is differentiated from astrocyte progenitor cells derived from human iPS cells, the human astrocyte cell population including at least 90% of human astrocytes, in which in the human astrocytes, a) CDKN2A is positive, b) at least one gene marker selected from the group consisting of IGFBP5, NNMT, HLA-DRB1, and HLA-DRB5 is positive, and c) an expression level of C3, which is standardized with GAPDH of a reference gene, is 0.05 copies/copies or less.

Provided herein are methods and compositions for a suicide gene approach comprising an expression vector comprising a cell cycle-dependent promoter driving the expression of a suicide gene. Also provided herein are methods to render proliferative cells sensitive to a prodrug after transplantation but avoids expression of the suicide gene in post-mitotic cells, such as neurons.

Provided herein are methods for obtaining populations of reprogrammed MO-homeostatic tolerogenic microglial cells. The methods include providing an initial population of monocytes, e.g., peripheral blood monocytes (PBMC), from a subject, and reprogramming the cells by maintaining the PBMC in culture ex vivo in the presence of a sufficient amount of transforming growth factor-beta (TGFβ) and interferon-gamma (IFNγ) for a time and under conditions sufficient for the cells to become M0-homeostatic tolerogenic microglia. Also provided are methods of use of these cells, e.g., for the treatment of neurodegenerative diseases associated with inflammation, e.g., Alzheimer's Disease (AD); Multiple Sclerosis (MS), e.g., progressive MS; and Amyotrophic Lateral Sclerosis

Disclosed herein are methods and compositions for the cryopreservation of stem cells, such as stem-cell derived retinal pigment epithelial cells, that have been seeded onto and cultured on a substrate, such as a polymeric substrate. Such cryopreserved stem cells are useful for cell therapies, such as treatment of ocular damage or disease.

Materials and methods for cell-mimetics having mechanical properties of biological neural axons are provided. A cell-mimetic device includes an array of fibers comprised of hexanediol diacrylate (HDDA) or an HDDA derivative, and at least one derivative of polyethylene glycol (PEG) selected from the group consisting of: PEG-acrylate, PEG-diacrylate, and any multi-arm PEG-acrylate.