Development of a Dielectrophoretic System for Rapid and Efficient Stem Cell Separation
Stem cells have been characterized and isolated based on a range of criteria, including surface proteins, morphology, and dye exclusion, mostly visualized by either histology or flow cytometry. Often a combination of techniques or markers are used to define a stem or progenitor cell, exemplified by multiple cytometric parameters commonly used to characterize hematopoietic or embryonic stem cells. Developmental studies and clinical application depends on characterization and isolation of stem cells from complex cell mixtures. With the exponentially growing knowledge of the multitude of stem and progenitor cells that exist in diverse and often complex organs or cultures, there has been a corresponding increase in the need for novel markers and separation technologies to precisely distinguish specific cell populations. Our proposal to develop a DEP system is significant in that it applies a novel technology to the problem of cell separation and characterization. DEP separates cells based on size and intrinsic electrical properties, and we propose can be developed as a stand-alone system as well as in conjunction with traditional separation or characterization technologies. Preliminary data are presented showing feasibility of DEP separation of cells in biological media and solutions with retention of cell viability. The specific goal of our research is to scale up existing prototype instruments and to develop methods separation methods based on intrinsic cellular properties as well as use of known surface epitopes as currently done in traditional histology and flow cytometry.
Cell separation and characterization is the basis for pinpointing stem cells within an organ or cell culture. Thus, the need to define and purify cells underlies the entire stem cell research and clinical enterprise. With exponentially growing knowledge of the multitude of stem and progenitor cells that exist in complex tissues and cultures has come an increased need for methods and markers to define and purify stem and progenitor cells. In part, this can be addressed by development of additional cell surface markers, as well as genomic or proteomic profiling. However, new separation technologies that introduce novel parameters, as proposed in this application, will contribute to solving the problem.
Preliminary data using small-scale instruments indicate feasibility of the technology. The goals are the development of a large-scale instrument and matching methods and reagents for stem and progenitor separation. Benefits to California would accrue through:
1. Support of the stem cell research enterprise.
2. Intellectual property that would be transferred to California companies.
3. Potential founding of new companies to develop commercializable versions of the instrument and/or technology.
California has many models for companies that offer cell separation technologies and instruments, including: Beckman Coulter, Becton Dickinson, Invitrogen, as well as many smaller companies that would be capable of developing commercial versions of the prototype instruments proposed in this application. Therefore, we anticipate that our goal of developing DEP cell separation technology will find applications in basic research, biotechnology and clinical settings through prototype instruments and through the production of commercial instruments.