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Provided is a method for producing sinusoidal endothelial cell-like cells (SEC-LCs) comprising steps of providing stem cells, which may be iPS cells, or angioblasts, and culturing the stem cells or angioblasts under conditions comprising; a) culturing the stem cells or angioblasts in the presence of bFGF; or b) culturing the stem cells or angioblasts in the presence of VEGF-A to produce endothelial cells followed by culturing the endothelial cells in the presence of a TGF-beta signaling inhibitor, cAMP signaling agonist, VEGF-C; or c) culturing the stem cells or angioblasts under hypoxic conditions, thereby producing SEC-LCs.

Provided is a method for identifying an agent for treating or ameliorating symptoms of cardiovascular diseases (CVDs), comprising (a) contacting candidate compounds with iPSCs bearing CVD risk 9p21.3 haplotype, which may be generated from PBMC of subjects, (b) examining the compounds-contacted iPSCs' differentiation into vascular smooth muscle cells ("VSMC differentiation"), and (c) examining VSMC differentiation of negative control cells, which are the same iPSCs not contacted with the candidate compounds; wherein an improved VSMC differentiation of iPSCs contacted with a specific candidate compound, relative to that of the negative control cells, identifies the specific candidate compound as an agent for treating or ameliorating symptoms of CVDs.

Provided is a method for in vitro production of photoreceptor precursor cells, comprising steps of; (a) 3D sphere culturing pluripotent stem cells, which may be iPS cells, to generate first spheres comprising eye early and late committed retinal neural progenitors (CRNPs); (b) monitoring sphere size until the first spheres reach an average size of about 300-500 pm in diameter; (c) disassociating the first spheres into first substantially single cells; (d) 3D sphere culturing the first substantially single cells to generate second spheres comprising photoreceptor precursor cells (PRPCs); (e) monitoring sphere size until the second spheres reach an average size of about 300-500 pm in diameter; (f) disassociating the second spheres into a second plurality of substantially single cells; (g) 3D sphere culturing the second plurality of substantially single cells to generate third spheres comprising postmitotic PRPCs; and (h) optionally, further differentiating the postmitotic PRPCs into photoreceptor-like cells.

Provided is a method for generating beta cells from pluripotent stem cells (PSCs) comprising steps of; (a) culturing PSCs in a medium comprising a GSKP inhibitor and a BMP inhibitor for 4-10 days to obtain definitive endoderm (DE) cells, (b) culturing DE cells in a medium comprising a GSKP inhibitor, a BMP inhibitor, and a Hedgehog inhibitor for 2-6 days to obtain pancreatic progenitor 1 (PP1) cells, (c) culturing PP1 cells in a medium comprising a BMP signaling inhibitor for 2-6 days to obtain pancreatic progenitor 2 (PP2) cells, (d) culturing PP2 cells in a medium comprising T3, and a Hedgehog inhibitor, a retinoic acid-signaling activator, a Notch inhibitor, and a TGF-beta inhibitor for 5-10 days to obtain endocrine progenitor (EP) cells, and (e) culturing EP cells in growth-factor-free, defined medium for 10-20 days to obtain beta cells.

Provided is a method for producing a CD3-positive or CD3 and CD197-double positive cell comprising a step of culturing a CD3-positive cell which may be derived from an iPS cell with a CD3/TCR complex agonist, fibronectin or its derivative, and a CD30 agonist.

Provided is a method for producing myocardial cells comprising steps of: (1) a step of subjecting iPS cells to spheroid culture using a culture substratum having multiple compartments so that multiple embryoid bodies can be spheroid-cultured in an individually separated state, thereby forming multiple embryoid bodies, and (2) a step of subjecting the multiple embryoid bodies produced in step (1) to 3D suspension culture to induce differentiation of the multiple embryoid bodies into myocardial cells.

Provided is an in vivo delivery system of DNA modifying enzymes comprising stem cell (e.g., iPS cell)-derived extracellular vesicles which are defined by the expression of surface markers and a portion of which contains mRNA of hybrid nucleases, recombinases, integrase, resolvases/invertases or transposases.

Provided is a pharmaceutical composition for use in treating or diminishing debilitating conditions in a subject, comprising at least 105 to 2x107 human stem cells (e.g., iPS cells, mesenchymal stem cells, and so on) per kilogram bodyweight of the subject, wherein the human stem cells are enriched with frozen-thawed healthy functional exogenous mitochondria and wherein the debilitating conditions are selected from the group consisting of aging, age-related diseases and the sequalae of anti-cancer treatments.

Provided is an isolated hypoimmune cardiac, endothelial, neuronal, islet, or retinal pigment cell differentiated from a hypoimmune induced pluripotent stem cell (HIP cell). HIP cells have, for example, a reduced or eliminated endogenous b-2 microglobulin (B2M) gene activity, a reduced or eliminated endogenous class II transactivator (CIITA) gene activity, and increased CD47 expression.

Provided is a method for producing a pluripotent stem cell product comprising steps of; (1) culturing pluripotent stem cells in an undifferentiated cell maintenance culture medium, (2) suspending the pluripotent stem cells cultured in (1) in a first cell suspension medium containing a ROCK inhibitor, (3) replacing the cell suspension medium in the suspension obtained in (2) by a cryopreservation medium to produce a cell dispersion composed of the cryopreservation medium and the pluripotent stem cells dispersed in the cryopreservation medium, and (4) filling the cell dispersion obtained in (3) in an air-tight container in an air-tight state.

Provided is a method for inducing a glomerular podocytes in vitro comprising steps of (a) culturing a nephron precursor cell, which may be derived from mammalian iPS cells, in a medium containing a Wnt agonist and a ROCK inhibitor, (b) culturing the cell obtained in (a) in a medium containing FGF, and (c) culturing the cell obtained in (b) in a medium including FGF, wherein at least one medium in the step (b) or (c) contains a TGFβ signal pathway inhibitor.

Provided is a non-invasive evaluation method of genomic abnormality in pluripotent stem cells or cells differentiated from the same, using as an index the level of at least one selected from the group consisting of deoxycytidine, kynurenine, putrescine, alanine, cysteine, cystathionine, or threonic acid in the cell culture supernatant.

Provided is a method for detecting undifferentiated cells, comprising a step of detecting the expression level and/or the promotor activity of at least one gene selected from the group consisting of ESRG, VSNL1, THY1, SFRP2, SPP1, USP44, and CNMD.

Provided is a method for generating a 3D organoid comprising a population of hindbrain cells comprising a heterogeneous population of interneurons, the method comprising steps of treating a population of mammalian pluripotent stem cells (PSCs) in vitro with a WNT signaling pathway activator and culturing the population of PSCs in a neural induction medium comprising a retinoic acid signaling pathway activator, a sonic hedgehog signaling pathway activator and a Notch signaling pathway inhibitor.

Provided is a method for accelerating cell lineage conversion (in a preferred embodiment, reprogramming a somatic cell into an iPS cell) comprising steps of transfecting a cell with a composition comprising at least one synthetic mRNA encoding an engineered chimeric transcription factor (in a preferred embodiment, based on Oct4 and/or Sox2 and/or Nanog) fused with a heterologous peptide sequence derived from the C-terminal transactivation domain (TAD) of Gal4.

Provided is a three-dimensional tissue product derived from human iPSCs comprising functionally matured retinal pigment epithelial (RPE) cells, a portion of three-dimensional neural retina (3DNR), an additional biocompatible component, and a biocompatible scaffold, wherein the 3DNR, the RPE cells, and the additional biocompatible component are physically and functionally integrated to form a complex containing a layer of neural retina and an underlying layer of RPE cells, wherein the 3DNR comprises (i) undifferentiated pseudostratified neural retinal epithelium, (ii) laminated neural retina tissue including all retinal layers and their corresponding retinal precursor cell types, or (iii) advanced differentiated retinal tissue including an outer nuclear layer (ONL) and a bipolar cell layer (BCL), wherein the ONL could be rod-enriched, cone- enriched, or any combination thereof, wherein the additional biocompatible component comprises a natural or synthetic compound in a liquid or gel form that provides an appropriate biomechanical environment for cell survival and function, allows manipulation of the product, or both, and wherein the RPE cells are grown on top of said biocompatible scaffold prior to integration with the 3DNR, and wherein the 3DNR is positioned on top of the RPE cells.

Provided is a method for generating lung progenitor cells comprising steps of (a) producing anterior foregut endoderm cells from mammalian pluripotent stem cells (PSCs), (b) culturing the anterior foregut endoderm cells in a suspension culture to generate at least one lung bud organoid (LBO), (c) embedding the LBO within a 3D matrix, (d) culturing the embedded LBO to form branched LBO (BLBO), and (e) dissociating the LBO or BLBO and culturing the dissociated LBO or BLBO on feeder cells in a culture medium.

Provided is a method for generating an enriched population of monohormonal cells from pluripotent stem cells comprising steps of exposing pluripotent stem cells sequentially to (a) CHIR99021 and Activin A, (b) Activin A, (c) FGF7, (d) retinoic acid, LDN 193189 and SANT 1, and (e) retinoic acid, LDN193189, SANT1, EGF and FGF2, wherein at least 50% of the cells in the generated population are monohormonal and express insulin, but not glucagon, somatostatin, or ghrelin.

Provided is a method for producing conjunctival epithelial cells, conjunctival goblet cells and conjunctival epithelial stem/precursor cells in which a pluripotent stem cell colony is cultured and differentiated in a culture medium contatining EGF signal activator.

The present invention addresses the problem of establishing a method for proliferating human noroviruses (HuNoV) in vitro, while avoiding ethical issues accompanying the conventional method including a need to treat the cells with bile. Specifically, the present invention relates to a method for proliferating HuNoV comprising a step of infecting an iPS cell-derived intestinal epithelial cell with HuNoV, and a screening method of a HuNoV-proliferation inhibitor using said method.

Provided is a method for producing stem cell-derived lacrimal gland tissue and is characterized by isolating cells that are positive to both SSEA4 and CD104 from a self-formed ectodermal autonomous multi-zone (SEAM) cell population derived from pluripotent stem cells, and three-dimensionally culturing the obtained cells in a culture medium containing EGF and a ROCK inhibitor, to obtain a cell population that expresses lacrimal gland-associated proteins.

Provided is a method for inducing differentiation of pluripotent stem cells into cerebrovascular endothelial cells capable of forming tight junctions that are stable and can be maintained for a long period of time, comprising steps of; (1)cultivating pluripotent stem cells in the absence of FGF2 and reducing undifferentiation, and (2)differentiating the cells obtained in step (1) into brain microvascular endothelial cells, said step including cultivation in the presence of FGF2, retinoic acid and a TGF-β inhibitor.

Provided is a method for producing pancreatic β cells, comprising steps of; (a) producing posterior foregut (PFG) cells by culturing primitive gut tube (PGT) cells derived from pluripotent stem cells, in the presence of a protein kinase C (PKC) activator, (b)producing pancreatic precursor (PP) cells from PFG by culturing in the presence of retinoic acid or an analog thereof, (c) producing pancreatic endocrine precursor (EP) cells from PP by culturing in the presence of a Notch signal inhibitor and a ROCK signal inhibitor (or in a medium containing nicotinamide), and (d) producing pancreatic β cells from EP by culturing in the presence of an insulin receptor signal activator, transferrin, and selenious acid.

Provided is a medium for culturing and/or maintaining stem cells and stem cell-derived differentiated cells, which contains any one or a plurality selected from the group consisting of liarozole, piogliazone, silibinin, and chrysin as differentiation-controlling compound(s). The differentiation-controlling compound inhibits survival of undifferentiated stem cells without inducing apoptosis of differentiated cells, thus making it possible to obtain a population of differentiated cells with no incorporation of undifferentiated stem cells.

Provided is a method for producing a population of neural retinal progenitors (NRPs) comprising steps of; (a) obtaining a starting population of human iPSCs, (b) culturing the cells in a first retinal differentiation medium (RD1) comprising a BMP inhibitor to further differentiate the cells to anterior neuroectoderm cells, (c) inducing retinal differentiation of the anterior neuroectoderm cells by culturing the cells in a second retinal differentiation medium (RD2) essentially free of BMP inhibitors to form retinal progenitor cells (RPCs), and (d) culturing the RPCs in a retinal maturation (RM) medium to produce NRPs. Also provided is a method for producing a population of photoreceptor precursor cells (PRPs) from NRPs obtained by the above method by further culturing in photoreceptor precursor induction medium (FDSC) comprising a g-secretase inhibitor and FGF for a period of time sufficient to produce a population of PRPs.

Provided is an in vitro or ex vivo method of producing a population of human pluripotent stem cell derived macrophages with an altered phenotype, comprising a step of modulating the activity of KLF1 in the population. The inventors have shown that KLF1 activation programs iPS cell-derived macrophages into cells that have an erythroblastic island-like phenotype and is able to support the maturation and enucleation of differentiating erythroid cells.

The present invention relates to a composition for inducing dedifferentiation from somatic cells to iPS cells and a method of inducing dedifferentiation using the same, wherein the composition or the method increases the dedifferentiation efficiency by stimulating CXC chemokine receptor 2 (CXCR2), which is a receptor on human somatic cells.

Provided is a promoter that induces differentiation of pluripotent stem cells into insulin-producing cells, which comprises a compound having an LXR agonist-action. In a preferred embodiment, the said compound is an LXR endogenous ligand such as 22(R)-hydroxycholesterol, 24(S)-hydroxycholesterol, 24(S), 25-epoxycholesterol, or 27-hydroxycholesterol.

Disclosed is a method for enhancing stem cell differentiation into a beta cell by using one or more epigenetic modification compound(s) which is selected from a group consisting of a DNA methylation inhibitor, a histone acetyltransferase inhibitor, a histone deacetylase inhibitor (in a preferred embodiment, KD5170, MC1568, or TMP195), a histone methyltransferase inhibitor (in a preferred embodiment, DZNep, GSK126, or EPZ6438), or a bromodomain inhibitor.

Provided is a method for culturing pluripotent stem cells while maintaining undifferentiated state, comprising a step of culturing the cells on a substrate coated with sulfated hyaluronic acid having a degree of sulfation of 7% or greater in terms of S content.