A method of enhancing osteogenic differentiation using vitamin D treatment is disclosed. The method utilizes the combined effect of vitamin D treatment and flow-induced shear stress in a modified perfusion bioreactor to treat the bone defect. The method comprising the steps of: isolating adipose tissue from a subject by liposuction; separating adipose-derived stem cells from the adipose tissue; pre-treating the separated adipose-derived stem cells for a predefined time 20 to 40 minutes with vitamin D3; seeding the pre-treated stem cells onto one or more scaffolds; washing of unattached stem cells from the scaffolds after a predefined time of 20 to 30 minutes; culturing the stem cell seeded scaffold by utilizing a modified perfusion bioreactor to form a tissue-engineered construct, where flow induced shear stress is applied, and implanting the tissue-engineered construct into the subject without a need to obtain autologous bone graft.

A self-replicating RNA for inducing somatic differentiation of unmodified adult stem cells is described. Methods of differentiating unmodified adult stem cells into functional beta-like cells are also described, as well as compositions, tissues and devices containing such cells. The method requires inducing sequential expression of PDX1 before NGN3, and NGN3 before MAFA in these stem cells to form reprogrammed beta-cells. Self-replicating RNAs are provided and introduced into the adult stem cells to induce the sequential expression. Methods of treating diabetes are also provided, comprising obtaining stem cells, preferably from a patient with diabetes, inducing sequential expression of PDX1>NGN3>MAFA, in said stem cells to form reprogrammed beta-cells, and introducing said reprogrammed beta-cells into a pancreas of said patient.

An object of the present invention is to provide a method for producing insulin-producing cells having sufficient glucose responsiveness from mesenchymal stem cells, an insulin-producing cell having sufficient glucose responsiveness, a cell structure containing the insulin-producing cell, and a pharmaceutical composition. According to the present invention, there is provided a method for producing an insulin-producing cell from a mesenchymal stem cell, including (a) a step of producing a cell structure by incubating a plurality of biocompatible macromolecular blocks and a plurality of mesenchymal stem cells, and (b) a step of culturing one or more of the mesenchymal stem cells before the incubation in the step (a), the mesenchymal stem cell in the incubation in the step (a), or the cell structure produced in the step (a) in a medium containing the GLP-1 receptor agonist, and (c) a step of culturing the cell structure obtained in the step (a) or the step (b) in a medium containing the water-soluble vitamin and the hepatocyte growth factor.

Cocktails of chemical inducers of neuron-like properties (CINP) is provided, which includes cAMP agonists, neurogenic small molecules, glycogen synthase kinase inhibitors, TGF receptor inhibitors, and BET family bromodomain inhibitors and optionally, a selective inhibitor of ROCK or p38 MAPK. These cocktails are used in a method of inducing neuron-like properties in partially or completely differentiated non-neuronal cells. The method includes contacting cells of a first type (non-neuronal) with the CINPs for a sufficient period of time to result in reprogramming the cell into cells of a second type having neuron-like characteristics (CiNs). Isolated chemically induced neurons (CiNs) can be used in a number of applications, including but not limited to cell therapy.

A method of generating a population of cells useful for treating a brain disorder in a subject is disclosed. The method comprises contacting mesenchymal stem cells (MSCs) with at least one exogenous miRNA having a nucleic acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 15-19 and 27-35, thereby generating a population of cells and/or generating neurotrophic factors that may provide important signals to damaged tissues or locally residing stem cells. MSCs differentiated by miRs may also secrete miRs and deliver them to adjacent cells and therefore provide important signals to neighboring endogenous normal or malignant cells.

The present invention relates to a method of treating a subject having drug-resistant cancer, comprising administering a composition comprising exosomes derived from differentiating stem cells as an active ingredient. The exosomes isolated differentiating stem cells according to the present invention have an excellent expression rate of bioactive factors affecting differentiation and have an effect of facilitating reprogramming of cancer stem cells and differentiating them into cancer cells with weakened drug resistance.

The present invention provides a method for producing a unilocular adipocyte including inducing differentiation into unilocular adipocytes of mesenchymal cells having differentiation potency into adipocytes by culturing the mesenchymal cells in suspension in a liquid medium composition capable of culturing cells or tissues in suspension, wherein the liquid medium composition contains a polymer compound having an anionic functional group that binds via a divalent metal cation to form a structure capable of suspending cells or tissues, and the method wherein the polymer compound is polysaccharide, preferably polysaccharide containing a glucuronic acid moiety, more preferably deacylated gellan gum, diutan gum or xanthan gum or a salt thereof.

Methods of differentiating unmodified adult stem cells into functional beta-like cells are provided, as well as compositions, tissues and devices containing such cells. The method requires inducing sequential expression of PDX1, NGN3, and MAFA in these stem cells to form reprogrammed beta cells. Methods of treating diabetes are also provided, comprising obtaining stem cells, preferably from a patient with diabetes, inducing sequential expression of PDX1, NGN3, and MAFA, in said stem cells to form reprogrammed beta cells, and introducing said reprogrammed beta cells into a pancreas of said patient. Alternatively, it may be possible to inject such cells systemically, if the cells are targeted for the pancreas. In yet another embodiment, the reprogrammed beta cells are placed into an artificial pancreas that is surgically placed or injected into the patient.

A method of generating a population of cells useful for treating a brain disorder in a subject is disclosed. The method comprises contacting mesenchymal stem cells (MSCs) with at least one exogenous miRNA having a nucleic acid sequence at least 90% identical to a sequence selected from the group consisting of SEQ ID NOs: 15-19 and 27-35, thereby generating a population of cells and/or generating neurotrophic factors that may provide important signals to damaged tissues or locally residing stem cells. MSCs differentiated by miRs may also secrete miRs and deliver them to adjacent cells and therefore provide important signals to neighboring endogenous normal or malignant cells.

A method of obtaining a pancreatic multipotent or unipotent cell including providing a cell of a first type which is not a pancreatic multipotent or unipotent cell; contacting the cell of a first type with an agent capable of remodeling the chromatin and/or DNA of the cell; transiently increasing expression of at least one pancreatic multipotent or unipotent gene regulator in the cell of a first type, to a level at which the at least one pancreatic multipotent or unipotent gene regulator is capable of driving transformation of the cell of a first type into the pancreatic multipotent or unipotent cell; and placing or maintaining the cell in a pancreatic cell culture medium and maintaining intracellular levels of the at least one pancreatic multipotent or unipotent gene regulator for a sufficient period of time to allow a pancreatic multipotent or unipotent cell to be obtained.