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.

The disclosure relates to a method of reprogramming one or more somatic cells, e.g., partially differentiated or fully/terminally differentiated somatic cells, to a less differentiated state, e.g., a pluripotent or multipotent state. In further embodiments the invention also relates to reprogrammed somatic cells produced by methods of the invention, to chimeric animals comprising reprogrammed somatic cells of the invention, to uses of said cells, and to methods for identifying agents useful for reprogramming somatic cells.

Provided is a method of generating iPS cells. Specifically, provided is a method of generating iPS cells having a rearranged -TCR gene. Also provided is a cell population including the generated iPS cells. The method includes stimulating collected blood cells with IL-2 and a bisphosphonate, and then introducing cell reprogramming factors through use of a Sendai virus (SeV) vector. According to the method of the present invention, iPS cells having a rearranged -TCR gene can be effectively generated. In particular, the method may be free of a step of treating the blood cells with an antibody before the step of stimulating blood cells with any one kind or a plurality of kinds of interleukins selected from IL-2, IL-15, and IL-23, and a bisphosphonate. In addition, iPS cells generated by the method of the present invention can be differentiated into desired cells by differentiation induction treatment.

The present invention provides a production method of iPS cell, including a step of introducing the following (1) and (2): (1) an episomal vector containing a nuclear reprogramming factor; and (2) an episomal vector containing EBNA-1, which is different from (1), into a somatic cell, as well as a method for improving iPS cell establishment efficiency. The present invention also provides an agent for improving iPS cell establishment efficiency, which contains an episomal vector containing a nucleic acid encoding EBNA-1, and a kit for producing an iPS cell further containing an episomal vector containing a nucleic acid encoding a nuclear reprogramming factor.

The nuclear reprogramming of somatic cells with mRNA encoding reprogramming factors is shown to be greatly accelerated by activation of innate immune responses in the somatic cell. Methods of activating innate immunity include activation of PKR, of toll-like receptors, e.g. TLR3, etc. In some embodiments the mRNA provides the activator of innate immunity.

A pharmaceutical composition includes a cell-free conditioned medium (CM) or extract or concentrate thereof obtained from a mammalian cell culture medium comprising a cultured substantially homogenous cancerous mammalian cell population. At least a portion of the cancerous mammalian cell population is contacted by at least one small molecule cell growth signaling pathway activator before being cultured in the cell culture medium.

Compositions of transcription factor cocktails including engineered SOX17 factor (eSOX17), together with one or more of OCT4, KLF4, and C-MYC, and vectors carrying one or more of these transcription factors suitable for delivery to donor somatic cells for generating induced expanded potential stem cells (iEPSCs) or induced neural stem cells (iNSCs) are provided. The disclosed compositions deliver reprograming of human somatic cells into a totipotency-like, or multipotency-like state with improved efficiency. Compositions of eSOX17-derived iEPSCs and iNSCs generated according to the described methods are also described. The methods engineer somatic cells to express one or more markers of pluripotency or neural stem cells. Methods of using iEPSCs or iNSCs in studying early human development and in cell therapy are also provided.

Methods are provided for the ex vivo reprogramming of microbiopsies of tissue such as from the skin, wherein the cells from the tissue are reverted to a state capable of extensive expansion in volume and when transplanted in vivo, of promoting scarless tissue regeneration at the site of administration.

A culture medium is provided which is capable of establishing expanded potential stem cell (EPSC) lines which resemble nave or ground state ES cells, but are also able to differentiate into placenta trophoblasts and the embryo proper. Methods are provided using the medium for the in vitro conversion and maintenance of cells, including pluripotent cells into EPSCs.

The present invention relates to artificial transcription factors (ATFs) that alter gene expression, including inducing pluripotency in cells or promoting the conversion of cells to specific cell fates. In particular, provided herein is a zinc-finger based ATF library that can be screened in cells by looking for expression of a specific gene (e.g., reporter expression), monitoring for cell surface markers or morphology, or via functional assays.