A method for producing hepatocytes from hepatoblasts is provided. The method includes the step of culturing the hepatoblasts in a medium containing a compound selected from the group consisting of pregnenolone and an adrenergic agonist. The hepatoblasts can be obtained by culturing endodermal cells in a medium containing DMSO, and the endodermal cells can be obtained by culturing pluripotent stem cells in a medium containing Activin A and a GSK-3 inhibitor. Accordingly, a method for producing hepatocytes from pluripotent stem cells is also provided by employing the method of the present invention.

The present invention relates to an in vitro method of generating cells capable of differentiating to a multicellular organoid unit that morphologically and/or functionally recapitulates invasive and/or ductal cell growth. The present invention further relates to a method of screening for an anti-migratory drug using a multicellular organoid unit obtained in the in vitro method. Additionally, the present invention relates to a culture medium and the respective use of said culture medium in any of said methods according to the present invention.

Provided are methods, apparatus, pharmaceutical compositions, and kits for treatment of a metabolic condition, including obesity and type 2 diabetes, by administration to a subject of a therapeutically effective amount of a cell or tissue preparation such as brown adipose microtissues or brown adipose tissue directly converted from white adipose tissue. Modified approaches to creating brown adipose tissue involve differentiation of explanted white adipose tissue and direct browning of white adipose tissue in a bioreactor rather than isolation and expansion of adipose stems cells or endothelial cells and formation and differentiation of 3D cell aggregates.

A method of cell culture comprising providing cells in a cell culture medium to start a cell culture process, and, maintaining at least one metabolite selected from 3-(4-hydroxyphenyl)lactate, 4-hydroxyphenylpyruvate, phenyllactate, indolelactate, indolecarboxylic acid, homocysteine, 2-hydroxybutyric acid, isovalerate and formate below a concentration C1 in the cell culture medium, wherein C1 is 3 mM and/or (ii) maintaining at least one amino acid selected from phenylalanine, tyrosine, tryptophan, methionine, leucine, serine, threonine and glycine below a concentration C2 in the cell culture medium, wherein C2 is 2 mM.

A method of cell culture comprising providing cells in a cell culture medium to start a cell culture process, and, maintaining at least one metabolite selected from 3-(4-hydroxyphenyl)lactate, 4-hydroxyphenylpyruvate, phenyllactate, indolelactate, indolecarboxylic acid, homocysteine, 2-hydroxybutyric acid, isovalerate and formate below a concentration C1 in the cell culture medium, wherein C1 is 3 mM and/or (ii) maintaining at least one amino acid selected from phenylalanine, tyrosine, tryptophan, methionine, leucine, serine, threonine and glycine below a concentration C2 in the cell culture medium, wherein C2 is 2 mM.