Tracking of Human Embryonic Stem Cell Differentiation and Migration

Funding Type: 
Tools and Technologies I
Grant Number: 
RT1-01130
Investigator: 
ICOC Funds Committed: 
$0
Public Abstract: 
During normal embryonic development, cells follow a beautifully coordinated migration and differentiation program (to form tissues, then organs and whole animals); many details of which remain a mystery. Likewise, human embryonic stem cells (hESCs) differentiate into many tissue types and form rudimentary structures. In culture, we see differentiation of hESCs into, e.g., mesodermal cells that then differentiate into beating masses of heart cells, blood vessel-like structures and rudimentary neuro-vascula-like bundles; indicating the ability to form many of the tissues present in animals. Detailed analyses of the behaviors of human embryonic stem cells (hESCs) have not been possible because the huge volume of information is difficult to acquire and impossible to analyze with current tools. Needed automated computerized tools are proposed here to acquire, organize and analyze the huge quantity of information that will detail the movements, differentiation/development, cell division and interactions of hESC. In the embryo, key differentiation signaling events occur as a result, of cell-cell interactions as well other triggers. Our proposed development of biosensors that function in living cells will greatly aid automated identification of key developmental changes. Understanding these events is will help scientists control them, generate purified populations of appropriate progenitors, and learn to treat patients for a broad range of diseases. We propose to develop stable fluorescent protein (FP) labeled cell lines, automated microscopy and software tools for tracking hESC migration and cell lineages during differentiation into various cell types and structures. The Specific Aims are to: 1. 1. Develop and disseminate stable FP-reporter cell lines to label important markers of differentiation and drug selectable for isolation of pure populations of cardiomyocytes and endothelial cells. 2. Develop and disseminate automated microscopy and analysisstandard operating procedures (SOPs) for long-term (hours to weeks) time-lapse imaging in 3D. 3. Develop and disseminate software for fully automated tracking of the migration, differentiation/lineage, division, and intercellular interactions of hESCs. 4. Test and validate tracking of stable cell lines in SA1to track the appearance of endothelial cells and cardiomyocytes in EBs. The software tools will be disseminated by [REDACTED], and the stable biosensor hESC cell lines will be placed in [REDACTED] Ste Cell Center; both will be disseminated to stem cell scientists at large. These new tools will enable scientists to perform rapid, in-depth studies of hESC development and differentiation; enabling them to rapidly discover, test, improve and implement techniques for directing differentiation into appropriate cells, ultimately for therapeutic use.
Statement of Benefit to California: 
Discovering stem cell-derived treatments for important diseases is scientifically challenging. We propose to enable scientists to obtain the information needed to more rapidly overcome many of these challenges by developing and disseminating: 1) living human embryonic stem cell (hESC) lines containing fluorescent protein biosensors of key points of development/differentiation and 2) time-lapse automated microscopy image acquisition and software for organizing, collating and simplifying analyses of the complex movement, development, division and interplay of hESCs in culture. During normal embryonic development, cells follow a beautifully coordinated migration and differentiation program (to form tissues, then organs and whole animals); many details of which remain a mystery. Likewise, human embryonic stem cells (hESCs) differentiate into many tissue types and form rudimentary structures. In culture, we see differentiation of hESCs into, e.g., mesodermal cells that then differentiate into beating masses of heart cells, blood vessel-like structures and rudimentary neuro-vascular-like bundles; indicating the ability to form many of the tissues present in animals. Detailed analyses of the behaviors of human embryonic stem cells (hESCs) have not been possible because the huge volume of information is difficult to acquire and impossible to analyze with current tools. Needed automated computerized tools are proposed here to acquire, organize and analyze the huge quantity of information that will detail the movements, differentiation/development, cell division and interactions of hESCs. In the embryo, key differentiation signaling events occur as a result, of cell-cell interactions as well other triggers. Our proposed development of biosensors that function in living cells will greatly aid automated identification of key developmental changes. Understanding these events is will help scientists control them, generate purified populations of appropriate progenitors, and learn to treat patients for a broad range of diseases. We propose to develop stable fluorescent protein (FP) labeled cell lines, automated microscopy and software tools for tracking hESC migration and cell lineages during differentiation into various cell types and structures. The software tools will be disseminated commercially, and the stable biosensor hESC cell lines will be placed in a public stem cell center; both will be disseminated to stem cell scientists at large. These new tools will enable scientists to perform rapid, in-depth studies of hESC development and differentiation; enabling them to rapidly discover, test, improve and implement techniques for directing differentiation into appropriate cells, ultimately for therapeutic use. These new tools will enable California take/maintain its lead in stem cell research, rapidly develop stem cell-enabled treatments of important diseases, and coincidently contribute to a vibrant economy.

© 2013 California Institute for Regenerative Medicine