Optimizing Sorting Flow Cytometry for Translational Stem Cell Therapies

Funding Type: 
Early Translational I
Grant Number: 
ICOC Funds Committed: 
Public Abstract: 
When stem cells are grown in the laboratory, they often grow into a variety of cell types, not just the type that would ultimately be useful in cell-based therapies. Our goal is to achieve nearly pure collections of stem cells from these cultures. To achieve this goal, we will improve and customize cell sorting using a technique called fluorescence activated cell sorting (FACS). This technique allows a researcher to separate cells based on measurements of shape, size and proteins present on their surfaces. This separation is done rapidly and with high precision, but has not been optimized for routine use with stem cells. Improvements in this technology can produce higher and more consistent yields of specific stem cells by purifying them from undesired types in a scalable and economical way. We will define conditions and optimize methods for purification to the level necessary to ensure transplant safety. Using FACS for stem cell sorting is a natural fit, as stem cell cultures produce closely related sub-populations that require careful analysis to separate. Stem cell sorting has remained an niche technology for purifying stem cell populations for a variety of reasons. FACS instruments are made to accommodate a wide variety of cell types, and are therefore complex, and FACS has not been thoroughly optimized for stem cells. Success with FACS can also be cell type dependent, with stem cells being highly variable and sensitive to stress. Among the significant challenges in the use of current FACS instruments for purifying stem cells for patient therapy, perhaps the most important is the cumbersome implementation of clinically suitable sorting due to concerns over sterility and patient-to-patient contamination. Improvements achieved in this proposal will address these issues and be made available as a central resource to guide researchers in successfully preparing and sorting stem cells. By combining novel instrument capabilities with FACS expertise, we seek to address the needs of the stem cell community to allow the routine separation of stem cell populations for research and clinical use.
Statement of Benefit to California: 
The work outlined in this proposal would benefit California by supporting its position as a world leader in stem cell related knowledge and technologies. This is a collaborative effort between a privately held California company and a local research institution to provide techniques, tools and technology for advanced stem cell research. The work will employ local researchers to carry out the daily experiments, it will require many locally produced reagents and supplies, and include major equipment purchases from California companies to complete the proposed aims. This California based spending would contribute to the $2.2 to $4.4 billion dollars in additional income and sales tax revenue predicted from Proposition 71*. In addition to boosting the California economy, this work will depend on local scientific expertise at both academic institutes and local companies, ensuring productive work and advances in the scientific fields of some of California’s brightest minds. In addition to providing valuable gains in scientific knowledge for the company and collaborating institute, the work will be made widely available to the larger California research community. This will include online web-based availability, and a plan for an in-depth training course hosted at a local institute for hands-on dissemination of the knowledge gained through this work. In turn, this training would ensure future generations of California scientists benefit from this work and allow them to rapidly advance future stem cell therapies. If stem cell therapies are to be realized, there is a need for rapid and effective methods for separating the cells of interest from the background. There is currently no clear solution to this problem, and that is what this proposal will address. Producing safe, effective cell purifications will more rapidly allow these promising therapies to enter into clinical use. These therapies could provide relief for a host of conditions from arthritis to heart disease, and are predicted to save the State of California between $3.4 and $6.9 billion health-care related dollars*. *Analysis Group Report, Economic Impact Analysis, Proposition 71 California Stem Cell Research and Cures Initiative, 2004.

© 2013 California Institute for Regenerative Medicine