A method of obtaining 3D cell structures including differentiated human hepatic cells. The method includes: a first step of culturing stem cell-derived or immortalized human hepatic progenitors in a non-adherent culture vessel, preferably a low or ultra-low attachment culture vessel; a second step of transferring the stem cell-derived or immortalized human hepatic progenitors to a culture medium including methacrylated gelatin (GelMa), thereby embedding the stem cell-derived or immortalized human hepatic progenitors in a GelMa matrix; and a third step of covering the GelMa matrix with culture medium and culturing the stem cell-derived or immortalized human hepatic progenitors embedded in the GelMa matrix, thereby obtaining 3D cell structures including differentiated human hepatic cells. Also, methods for engineering an artificial liver model or an artificial liver organ, and for assessing in vitro the metabolism, toxicity and/or therapeutic effects of a compound.

The present invention relates to a multicellular construct that includes cells and a scaffold. The scaffold is constituted by a layered composite that comprises a gelatin nonwoven containing gelatin as a main component, and a gelatin film containing gelatin as a main component and layered on one surface of the gelatin nonwoven, and the cells are present in at least one of a region on the surface of the gelatin nonwoven and a region inside the nonwoven. The multicellular construct can be produced by arranging the scaffold in a swollen state inside the culture vessel whose inner surface is in the dry state such that the gelatin film of the scaffold is in contact with the inner bottom surface of the culture vessel, dripping a cell suspension onto the gelatin nonwoven of the scaffold, and then culturing the cells. This makes it possible to provide a multicellular construct with high seeding efficiency in which desorption of cells from a scaffold is suppressed, a method for manufacturing the same, a kit for producing the same, and a method for evaluating a compound using the same.

Embodiments of this disclosure relate to bioinks and bioink compositions. These bioinks may be 3D printed into a hydrogel. The printed hydrogel may support primary cell and induced pluripotent stem cell attachment, proliferation, and spreading. Compounds in the bioink may be modified to incorporate chemical functionality, such as by chemical synthesis means. Incorporating chemical functionality may allow the incorporation of modified material as a component in the bioink. The modifications may allow chemical conjugation of a desired component. The desired component may maintain its cell interactive feature to aid in cell attachment and proliferation. Such incorporation may allow modulation of the bioprinted object's mechanical properties without interfering with cell adhesion.

Methods for improved handling of isolated canine umbilical cord mesenchymal stromal cells (UC-MSCs), including methods for expansion of canine UC-MSCs, cryopreservation and improved post-thaw viability using adherent plates, as well as standardized methods and kits for characterizing isolated canine UC-MSCs in a cell population. Methods for improved detachment or dissociation of adherent cells and new dissociation reagents comprising nattokinase are also disclosed.

The present specification provides: a method of isolation of a pure culture of vascular endothelial cells, the method capable of isolating homogeneous endothelial cells adhered to a matrix for a specific time in a cell line of an endothelial cell lineage differentiated from human pluripotent stem cells; a medium for maintaining characteristics of vascular endothelial cells, comprising high-purity vascular endothelial cells isolated through the method, 4 ng/ml to 6 ng/ml of FGF2, 5 ng/ml to 10 ng/ml of EGF, 10 ng/ml to 30 ng/ml of VEGF-A, 20 ng/ml to 50 ng/ml of ascorbic acid, and DMEM/F-12 as active ingredients; and a culture method comprising same.

Biocompatible macroporous microcarriers, including microcarrier beads, microspheres, capsules, microsponges, hydrogels and other matrix forms, appropriate for use in a shaking flask or bioreactor to culture cells are described herein that can be used to create an edible structure for consumption or research investigation. Biocompatible, macroporous microcarriers can be dissolved or remain in the final product. Biocompatible macroporous microcarriers are formed by saccharides that are cross-linked via chemical induction with agitated cryo-gelation. Cross-linked macroporous, saccharide-microcarriers are coupled to adherence factors that enable cell binding. Finally, the cells are attached to the microcarrier for proliferation.

Disclosed herein is a method for covalent immobilization of molecular compounds on a substrate surface, comprising the steps: Providing a substrate surface; Treating the substrate surface with a plasma at atmospheric pressure, thereby generating an activated surface site; Exposing at least the activated surface site, or some fraction of the activated surface site, to molecular compounds, thereby establishing a covalent bond between the molecular compounds and the substrate surface.

A cell culture substrate having a high cell-adhesion portion and a low cell-adhesion portion, wherein; an adhesiveness to a cell of the high cell-adhesion portion and an adhesiveness to a cell of the low cell-adhesion portion are different from each other, the adhesiveness to the cell of the high cell-adhesion portion to cells is higher than the adhesiveness of the low cell-adhesion portion to the cell; and the high cell-adhesion portion has a cell adhesion layer containing two or more kinds of cell adhesion substances on the surface.

This disclosure relates to endothelial and smooth muscle like vascular tissue produced from urine cells. In certain embodiments, the disclosure relates to methods of producing endothelial and smooth muscle like vascular tissue by exposing urine derived cells with ETV2 in a first growth media under conditions such that the cells are modified to form a pool of cells expressing increased levels of endothelium surface markers and thereafter exposing the pool of cells to a second growth media under conditions such that the cells are modified to form tissue containing cells expressing increased levels of smooth muscle surface markers in addition to the endothelium surface markers. In certain embodiments, the disclosure relates to using cells and tissues reported herein for the treatment of vascular, cardiac, and wound healing indications.

A method for producing a fibrin sheet containing at least one selected from the group consisting of cells and drugs in a fibrin gel, the method comprising: a step 1 of applying a fibrinogen solution containing at least one selected from the group consisting of cells and drugs and fibrinogen dropwise onto a surface of a substrate made of a gelatin hydrogel; a step 2 of adding thrombin to the fibrinogen solution on the surface of the substrate; a step 3 of placing a support film on and in contact with a top surface of the fibrinogen solution to which the thrombin has been added; a step 4 of forming a fibrin sheet containing the at least one selected from the group consisting of cells and drugs in a fibrin gel between the substrate and the support film by a reaction between the fibrinogen and the thrombin; and a step 5 of melting the substrate at a temperature not lower than a melting temperature of the gelatin hydrogel to separate, from the substrate, the fibrin sheet supported by the support film.