The present disclosure provides in vitro methods for determining the potency of a cell-based therapy or treatment. In alternative embodiments, provided are compositions, including products of manufacture and kits, and methods, comprising (or comprising use of) quantitative in vitro assays for determining the potency of cell-based therapies or treatments, including those used in the treatment of retinal degeneration.

Provided herein are methods of producing, compositions comprising and uses of oligodendrogenic neural progenitor cells (o-NPCs), made using a combination of PDGFR agonist and thyroxin or a thyroxin analogue. The method includes; obtaining ventralized neural progenitor cells (NPCs), the ventralized NPCs expressing Sox2, Nkx6-1, decreased level of Pax6 compared to unpatterned NPCs, and elevated expression of HoxA4 compared to unpatterned NPCs; culturing the ventralized NPCs for about 12 to about 16 days (days 26-40 of FIG. 7; days 12 to 27 of FIG. 10) in neural expansion media (NEM) supplemented with i) PDGFR agonist for the about 12 to about 16 days and ii) thyroxine or a thyroxine analogue for the latter about 7 to about 9 days, to produce o-NPC expressing Sox2 and Nkx2.2, decreased level of Pax6 and Nkx6.1 compared to ventralized NPCs and elevated level of HoxA4 and Olig2 compared to ventralized NPCs.

A cell culture article includes a substrate having a polymer coating that is conducive to colony passaging of cells cultured on the coating. Example polymer coatings are formed from polygalacturonic acid (PGA), alginate, or combinations thereof. Cells cultured on the polymer coating can be separated from the substrate as a colony or layer of cells by exposing the polymer coating to (i) a chelating agent, (ii) a proteinase-free enzyme, or (iii) a chelating agent and a proteinase-free enzyme.

Provided are a method for producing a spermatogenic stem cell-like cell from a primordial germ cell-like cell derived from an isolated pluripotent stem cell in vitro, the method including (1) a step of coculturing a primordial germ cell-like cell with a gonad somatic cell in suspension to give reconstituted testis, and (2) a step of culturing the obtained reconstituted testis at gas/liquid interface to induce a DDX4-positive and PLZF-positive cell in the reconstituted testis; and
a method for producing a GSC-like cell, including dissociating a spermatogenic stem cell-like cell obtained by the method from the reconstituted testis, and culturing the cell under conditions that can induce a germline stem cell from the spermatogenic stem cell.

The present invention relates to a method of producing expandable cultured brain cells. The brain cells are neurotrophic factor (NTF) positive. The expandable cultured brain cells are obtained by culturing a biopsy obtained from the cortical and/or subcortical brain region of a living subject. The biopsies can be obtained during neurosurgical procedures such as deep brain stimulation. The expandable cultured brain cells of the present invention are useful for the treatment of neurological diseases and other medical conditions.

A construct that supports cell attachment and alignment including a substrate that is incompatible with photolithography conditions, containing a physical pattern in at least part of one surface, the physical pattern optionally bearing a coating of a metal alkoxide, oxide or mixed oxide-alkoxide thereon and a Self-Assembled Monolayer of Phosphonate (SAMP) covalently attached thereto, which phosphonate contains functionality adapted for cell binding. The construct optionally also contains cells attached thereto. Also disclosed are methods of preparing such a construct.

The invention relates to a method of producing a retinal pigment epithelial cell from a human pluripotent stem cell, and a method of treating or preventing a retinal disease by using the produced cell. The retinal pigment epithelial cell is prepared by (a) inducing differentiation of a human pluripotent stem cell into a pigment cell by adhesion cultivation of a human pluripotent stem cell in a medium containing a Nodal signal inhibitor and a Wnt signal inhibitor in the absence of a feeder cell to give a culture containing the pigment cell, (b) subjecting the obtained culture to further adhesion culture to give a culture containing a pigment cell colony, and (c) isolating the pigment cell from the obtained culture and culturing the cell to give a retinal pigment epithelial cell.

The present invention relates to novel compositions and methods to produce 3D organ equivalents of the brain (i.e. mini-brains). The invention also relates to methods of using human induced pluripotent stem cells, a combination of growth and other soluble factors and gyratory shaking. Cells from healthy or diseased donors or animals can be used to allow testing different genetic backgrounds. The model can be further enhanced by using genetically modified cells, adding micro-glia or their precursors or indicator cells (e.g. with reporter genes or tracers) as well as adding endothelial cells to form a blood-brain-barrier.

A cell culture article includes a substrate having a polymer coating that is conducive to colony passaging of cells cultured on the coating. Example polymer coatings are formed from polygalacturonic acid (PGA), alginate, or combinations thereof. Cells cultured on the polymer coating can be separated from the substrate as a colony or layer of cells by exposing the polymer coating to (i) a chelating agent, (ii) a proteinase-free enzyme, or (iii) a chelating agent and a proteinase-free enzyme.

The present invention provides a cell culture substrate including a polymer having a lower critical solution temperature, the substrate including one or more inorganic materials selected from a water-swellable clay mineral and silica and further including an adhesive matrix in the substrate, in which the adhesive matrix is an extracellular matrix and/or an adhesive synthetic matrix. Furthermore, the invention is to provide a cell culture substrate in which the extracellular matrix is at least one selected from laminin, fibronectin, vitronectin, cadherin, and fragments thereof, and/or the adhesive synthetic matrix is poly[2-(methacryloyloxy)ethyl dimethyl-(3-sulfopropyl) ammonium hydroxide] or an oligopeptide-supporting polymer.