The invention relates to improved culture methods for expanding epithelial stem cells and obtaining organoids, to culture media involved in said methods, and to uses of said organoids.

The present invention provides an approach for the determination of the activation states of a plurality of proteins in single cells. This approach permits the rapid detection of heterogeneity in a complex cell population based on activation states, expression markers and other criteria, and the identification of cellular subsets that exhibit correlated changes in activation within the cell population. Moreover, this approach allows the correlation of cellular activities or properties. In addition, the use of modulators of cellular activation allows for characterization of pathways and cell populations. Several exemplary diseases that can be analyzed using the invention include AML, MDS, and MPN.

Diagnostic tests and methods to assess the characteristics of a patient's native stem cell populations. A reference cell source may be selected for testing, to determine the relative health and potency of the cell population derived from that cell source based on standardized testing protocols, and to use the results to evaluate the systemic stem cell health status of the patient across the many different cell populations residing in different tissues in the body.

Methods of isolating cardiac cells, including cardiac cells capable of regenerating cardiac tissue are provided. Compositions comprising cardiac cells, including cardiac cells capable of regenerating cardiac tissue are also provided. Methods of using cardiac cells, cardiac progenitor cells, including cardiac cells capable of regenerating cardiac tissue, are provided. Methods of identifying the prognosis of patients treated for heart disease and/or methods of predicting the regeneration of cardiac cells in a subject are also provided.

The invention discloses a screening method for the identification of new compounds for use in the treatment of cancer.

A test animal for identifying blood vessel transendothelial migration-affecting factors or agents is described. The test animal comprises an implanted tissue mimicking model with a composite vascular structure comprising pericytes on test animal vasculature. The test animal can be used in a method of identifying or evaluating a blood vessel transendothelial migration-affecting agent that includes administering the agent to a test animal and determining the effect of the agent on the level of blood vessel transendothelial migration into or out of the tissue model. The test animal can also be used in a method of identifying or characterizing a metastasis-facilitating factor that includes administering or inducing circulating tumor cells in a test animal in which one or more factors in the pericyte have been silenced, and determining if the level of tumor cell migration into the tissue model has increased or decreased.

A three dimensional cell culture and bioreactor system is provided. The system comprises one or more cell culture chamber. Each cell culture chamber comprises an inlet port and an outlet port in fluid communication with the cell culture chamber. The cell culture chambers may be segregated or in fluid communication with one another. The systems may be used to conduct drug efficacy test, isolate certain cell types from a complex tissue sample of multiple cell types, allow for the ex vivo culturing of patient tissue samples to help guide the course of treatment, and conduct co-culture experiments.

The present invention relates to a composition for repressing the sternness of stem cells, which comprises a material for regulating OCT4 modification. The material for regulating OCT4 modification according to the present invention may regulate the phosphorylation or methylation of OCT4 and inhibit Wnt signaling, thereby effectively reducing the sternness of various stem cells. Therefore, since it can be effectively used in inhibition of proliferation, recurrence and metastasis of cancer, and inhibition of resistance to an anticancer agent, and can reduce sternness even in normal stem cells, it is expected that the time for differentiation of embryonic stem cells into specific cells is shortened, and efficiency is increased.

Herein, we describe a direct in vitro method that identifies agents that are toxic against natural human tissue stem cells. We provide a novel schedule for culturing any cell population containing homologous tissue stem cells that allows the number and cell kinetics of tissue stem cells, transient cells, and terminally differentiated cells within the population to be monitored. Using the passage schedule together with determination of a growth curve for the population, one can determine whether or not an agent is toxic to tissue stem cells, or to transient cells and/or terminal cells. The same method can also be used to identify agents that act positively on tissue stem cells and the other specific cell types.

SUSD2 protein is used as a marker in identification, selection or separation of pancreatic internal secretion precursor cells and/or newborn pancreatic internal secretion cells; and a use of an mRNA, for encoding the SUSD2 protein, of a precursor protein as the marker in identification of the pancreatic internal secretion precursor cells and/or the newborn pancreatic internal secretion cells. Analysis of gene expression of pancreatic endoderm cells sourced by induced directional differentiation of human pluripotent stem cells finds the enrichment expression of a SUSD2 gene in the pancreatic internal secretion precursor cells and the newborn pancreatic internal secretion cells. A protein encoded by the SUSD2 gene is a receptor protein on cell membranes. Using the protein as the marker, the identification, the selection or the separation of the pancreatic internal secretion precursor cells and the newborn pancreatic internal secretion cells can be conducted.