The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed.

A method of producing skin-derived precursor cells, comprising culturing human-derived pluripotent stem cells in a differentiation-inducing medium containing an agonist of Wnt signaling to differentiate the pluripotent stem cells into skin-derived precursor cells:, a differentiation-inducing medium for differentiating human-derived pluripotent stem cells into skin-derived precursor cells, comprising an agonist of Wm signaling as a differentiation-inducing promoter; and a differentiation-inducing promoter for differentiating human-derived pluripotent stem cells into skin-derived precursor cells, comprising an agonist of Wnt signaling as an active ingredient.

The invention relates to improved methods for culturing an epithelial stem cell or an organoid comprising epithelial stem cells. The invention also relates to culture media suitable for use with said methods, organoids obtainable or obtained by said methods and uses of said culture methods, media and organoids in drug discovery and validation, toxicity assays, diagnostics and therapy.

A bioactive suspension derived from freshly disaggregated tissue is provided, as well as related methods of formulation and use. The bioactive suspension may comprise a cell-free supernate derived from epidermal and dermal tissue that has been enzymatically and mechanically disaggregated, then separated, and which may contain tissue regeneration factors known to speed healing. The bioactive suspension may further comprise genetically-modified treatment cells, wild type cells, or both, and may be combined with one or more scaffolding elements to form a bioactive suspension combination product suitable for treatment of a cutaneous defect. Synthetic bioactive suspensions and bioactive suspension combination products are also provided.

A biological system for generating and preserving a repository of personalized, humanized transplantable cells, tissues, and organs for transplantation, wherein the biological system is biologically active and metabolically active, the biological system having genetically reprogrammed cells, tissues, and organs in a non-human animal for transplantation into a human recipient, wherein the non-human animal does not present one or more surface glycan epitopes and specific sequences from the wild-type swine's SLA is replaced with a synthetic nucleotides based on a human captured reference sequence from a human recipient's HLA.

A method for producing an epithelial cell sheet, comprising culturing cells derived from oral mucosal epithelial cells on a substrate in a serum-free medium, wherein the serum-free medium comprises (i) EGF protein or KGF protein, (ii) B-27 supplement, and (iii) a ROCK inhibitor.

Methods for detecting somatic mutations in single cells are described. Specifically, the disclosure provides methods of identifying somatic mutations in individual cells, comprising: providing a compartment containing only one somatic cell; generating genomic DNA and mRNA sequencing reads from the somatic cell; identifying potential mutations from the genomic DNA and mRNA sequencing reads relative to a control sequence; discarding artifact differences in the sequencing reads, thereby identifying somatic mutations in the individual cells compared to a control sequence.

Provided herein are constructs of micro-aggregate multicellular, minimally polarized grafts containing Leucine-rich repeat-containing G-protein coupled Receptor (LGR) expressing cells for wound therapy applications, tissue engineering, cell therapy applications, regenerative medicine applications, medical/therapeutic applications, tissue healing applications, immune therapy applications, and tissue transplant therapy applications which preferably are associated with a delivery vector/substrate/support/scaffold for direct application.

The growth factor profile, connective tissue matrix constituents, and immunoprivileged status of urodele extracellular matrix (ECM) and accompanying cutaneous tissue, plus the presence of antimicrobial peptides there, render urodele-derived tissue an ideal source for biological scaffolds for xenotransplantation. In particular, a biological scaffold biomaterial can be obtained by a process that entails (A) obtaining a tissue sample from a urodele, where the tissue comprises ECM, inclusive of the basement membrane, and (B) subjecting the tissue sample to a decellularization process that maintains the structural and functional integrity of the extracellular matrix, by virtue of retaining its fibrous and on-fibrous proteins, glycoaminoglycans (GAGs) and proteoglycans, while removing sufficient cellular components of the sample to reduce or eliminate antigenicity and immunogenicity for xenograft purposes. The resultant urodele-derived biomaterial can be used to enhance restoration of skin homeostasis, to reduce the severity, durations and associated damage caused by post-surgical inflammation, and to promote progression of natural healing and regeneration processes. In addition, the biomaterial promotes the formation of remodeled tissue that is comparable in quality, function, and compliance to undamaged human tissue.

Disclosed herein are synthetic leathers, artificial epidermal layers, artificial dermal layers, layered structures, products produced therefrom and methods of producing the same.