A high-throughput screening system is provided for optimizing the conditioning of patient-specific mesenchymal stem cells using a combinatorial set of biochemical factors, pharmacological inhibitors, and biomechanical forces. Also provided are generalized conditions for performing such conditioning. Cells made by these methods are also provided, in addition to cells having a mixed N endothelial cell/pericyte phenotype. These cells produce angiogenic growth factors and induce vascularization following implantation.

A method for evaluating the activity level of mesenchymal stem cells, and a method for culturing mesenchymal stem cells using the evaluation method in the field of culturing mesenchymal stem cells for regenerative medicine, and further, a method for producing a therapeutic agent for liver dysfunction and a therapeutic agent for liver dysfunction. This method for evaluating mesenchymal stem cell activity has an assay step for assaying the amount of adenylate kinase 4 (AK4) in the mesenchymal stem cells; and a determination step for determining the activity level of the mesenchymal stem cells from the assayed amount of adenylate kinase 4.

It is intended to provide a cancer stem cell and a method for preparing the same. The present invention provides a method for preparing a pluripotent cancer stem cell, comprising transferring Oct3/4, Sox2, Klf4, and c-Myc genes to an immortalized epithelial cell. The present invention also provides a pluripotent cancer stem cell as prepared by the above method.

The present invention discloses a method for establishing a colorectal cancer p73 reporter gene cell line, specifically including: first designing a site-specific sgRNA sequence of a p73 gene and cloning same into a plasmid PX459; integrating a homologous recombination sequence of the p73 gene and a green fluorescent protein DNA fragment (EGFP), and transforming the plasmid and the integrated fragment together into a colorectal cancer cell line HCT116 by electroporation; performing signal cell screening through a flow cytometer to obtain EGFP-expressing cells, and amplifying a monoclonal cell line; and identifying a positive p73 reporter gene cell line through PCR identification and Western blot, among screened EGFP-expressing cell lines. The colorectal cancer cell line p73 gene and the EGFP are co-expressed, and the expression level of the EGFP is highly consistent with that of the p73 gene. Therefore, the expression level of the p73 gene can be accurately determined by detecting changes in the expression level of the EGFP. The method for establishing the cell line in the present invention is simple, easy to implement, high in efficiency and precise in gene site positioning.

The present disclosure provides multipotent endoderm spheroid progenitor cells, liver organoids, intestinal organoids, and pancreatic spheroids, and methods for producing same from stem and progenitor cells.

Method for producing a primary culture of cancer cells, comprising: (a) fragmenting cancer tissues derived from a living body and removing impurities from the fragmented cancer tissues, (b) subjecting the tissue masses obtained in (a) to suspension culture, and (c) subjecting the culture obtained in (b) to adherent culture.

The invention provides a natural killer cell, NK-92, modified to express an Fc receptor on the surface of the cell, such as CD16 (FcRIII-A), or other Fc or Fr receptors. The modified NK-92 cell can be further modified to concurrently express an associated accessory signaling protein, such as FcRI-, TCR-, or to concurrently express interleukin-2 (IL-2) or other cytokines. Additional methods are disclosed for various assays, assessments, and therapeutic treatments with the modified NK-92 cells.

A 2D organoid for infection and growth culture of human diarrhea virus, obtained by: (1) obtaining a 3D organoid by three-dimensionally culturing a human intestinal epithelial stem cell, a human intestinal epithelial cell, or a tissue containing at least any of those cells on an extracellular matrix; and (2) dispersing the 3D organoid to prepare a single cell, and monolayer-culturing the single cell on an extracellular matrix to obtain the 2D organoid having a monolayer structure in which epithelial cells contain differentiated trophoblastic cells and goblet cells and configured as human intestinal lumen having a monolayer structure.

Disclosed herein are bioreactor systems and methods of utilizing said systems.

The present invention relates to microfluidic fluidic systems and methods for the in vitro modeling diseases of the lung and small airway. In one embodiment, the invention relates to a system for testing responses of a microfluidic Small Airway-on-Chip infected with one or more infectious agents (e.g. respiratory viruses) as a model of respiratory disease exacerbation (e.g. asthma exacerbation). In one embodiment, this disease model on a microfluidic chip allows for a) the testing of anti-inflammatory and/or anti-viral compounds introduced into the system, as well as b) the monitoring of the participation, recruitment and/or movement of immune cells, including the transmigration of cells. In particular, this system provides, in one embodiment, an in-vitro platform for modeling severe asthma as Severe Asthma-on-Chip. In some embodiments, this invention provides a model of viral-induced asthma in humans for use in identifying potentially effective treatments.