Systems and methods to detect stem cell stress and uses thereof
AssigneeWayne State University
InventorsDaniel A. Rappolee, Elizabeth Puscheck
Life of this patent
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Abstract
Systems and methods (S/M) to detect stress in stem cells are described. The S/M, including modified stem cells, assays and high throughput screens, can be used to identify compounds or other potential stressors that can negatively affect development potential.
Claims20
A method of detecting stress-induced differentiation in embryonic stem cells (ESCs) comprising:
- obtaining ESCs genetically modified to express a first reporter gene operably linked to a promoter of platelet-derived growth factor A (Pdgfra) gene;
- measuring expression level of the first reporter gene in the genetically modified ESCs;
- comparing the expression level of the first reporter gene to a reference first reporter gene expression level;
- and
- detecting that the genetically modified ESCs have undergone stress-induced differentiation when the measured first reporter gene expression level is statistically significantly increased as compared to the reference first reporter gene expression level,
- wherein the reference first reporter gene expression level is derived from corresponding genetically modified ESCs that have not undergone stress-induced differentiation, and
- wherein the genetically modified ESCs are in the presence of at least one of leukemia inhibitory factor (LIF) or fibroblast growth factor 4 (FGF4).
A method of identifying an agent that causes embryonic stem cells (ESCs) to differentiate in the presence of at least one of leukemia inhibitory factor (LIF) or fibroblast growth factor 4 (FGF4), the method comprising:
- contacting ESCs that are in the presence of LIF or FGF4 with an agent;
- measuring expression level of platelet-derived growth factor A (Pdgfra) from the contacted ESCs;
- comparing the measured Pdgfra expression level to a reference Pdgfra level;
- and
- identifying the agent as causing ESC to differentiate in the presence of LIF or FGF4 when the measured Pdgfra expression level is statistically significantly increased as compared to the reference Pdgfra level,
- wherein the reference Pdgfra level is derived from ESCs that have not undergone differentiation.
A method of detecting stress-induced differentiation in embryonic stem cells (ESCs) comprising:
- measuring platelet-derived growth factor A (Pdgfra) promoter activity level from ESCs that are in the presence of at least one of leukemia inhibitory factor (LIF) or fibroblast growth factor 4 (FGF4);
- comparing the measured Pdgfra promoter activity level to a reference Pdgfra promoter activity level;
- and
- detecting that the ESCs have undergone stress-induced differentiation when the measured Pdgfra promoter activity level is statistically significantly increased as compared to the reference Pdgfra promoter activity level,
- wherein the reference Pdgfra promoter activity level is derived from ESCs that have not undergone stress-induced differentiation.
Dependencies are inferred from the claim text.