Tools and Technologies I
We are proposing to optimize and scale up a highly advanced (microfluidic) cell culture system into manufacturable form. This system will allow researchers to: Identify stem cell culture and differentiation conditions Identify genes and small molecules effecting stem cell self-renewal and differentiation, and Identify genes and small molecules involving or effecting reprogramming of differentiated cells. ...much more rapidly and efficiently than they have been able to in the past. Reprogramming a patient's own differentiated cells (e.g. skin cells) into stem cells overcomes the ethical and immunological barriers to theraputic usage which are present with the use of embryonic stem cells. These stem cells can be used in cell based therapy, tissue or organ repair, and potentially even organ reconstruction. Understanding what controls stem cells to differentiate into a desired type of cell helps directly in the development of theraputic applications. Thus, this tool will help both to determine conditions to convert differentiated cells into stem cells, and to develop therapies using the resulting stem cells.
Statement of Benefit to California:
The system proposed here will allow us to Identify stem cell culture and differentiation conditions Identify genes and small molecules effecting stem cell self-renewal and differentiation, and Identify genes and small molecules involving or effecting reprogramming differentiated cells. These capabilities will accelerate stem cell research in California. Since the grant will support work done in South San Francisco and San Diego, and the end result may be the creation of a commercial product, there is a direct economic multiplier effect for the resources invested. More importantly, the identification of conditions which enable the reprogramming of differentiated cells will enable new therapies. Patient specific pluripotent stem cells can be used in cell based therapy, tissue or organ repair, and even organ reconstruction. The availability of powerful tools in California will help ensure that these new therapies are pioneered in California, leading both to job creation and the availability of the most advanced medical care in the world for California citizens.
This application represents a collaboration between two biotech companies, and focuses on translation of existing microfluidic cell culture arrays (developed by the applicants) into a manufacturable product. The device concept was developed in academia, using 96-well culture chambers that can be fed with arbitrary time-varying combinations of reagents, 16 reagents are proposed here. High throughput screening of libraries to identify factors that can control various aspects of stem cell fate is an important area of research, but not yet widespread, and here the applicants choose to focus most on reprogramming of somatic cells to induced pluripotent stem (iPS) cells. The potential impact of the work is very high because a lot of conditions can be tested on the same cells, and the technology developed in this work could be used to test not only combinations of signals, but automatically test the dynamics or kinetics of compound additions. Furthermore, one reviewer felt that the project will have a direct economic impact on the stem cell industry in California as the product should be quickly commercialized. The technology is elegant, and the plan to move the chip they have already developed into a commercial product is sound. The applicant describes plans to develop the prototype instrumentation necessary for commercialization, which requires chip redesign, a new carrier and new software to interface with a commercial controller that will also be developed as part of this project. One of the companies applying is a leader in commercializing microfluidic technology, and their multilayer soft lithography technology is entirely appropriate for the proposed application. The use of a chip carrier and the focus on user friendliness are also positives of this proposal. Success of the development phases of the research will be documented by carrying out culture experiments that are detailed in the literature. The proposed biological testing is not as strong. The applicants do not explain fully how they will use the features of their technology, and the plans do not take full advantage of microfluidic cell culture arrays, to push the technology to provide competitive advantages over multiwell plates. As described, the small-scale screens proposed could just as easily be performed in traditional multiwell plates. For example, at one point in which pulses of exposure would be a logical parameter to examine, the applicants only discuss day-long exposures to various conditions. Along these lines, few details of the conditions to be tested are included, and there is no discussion of the anticipated number of conditions to be screened to ensure that they are reasonable. The applicants also do not sufficiently acknowledge the anticipated challenges involved in growing human embryonic stem cells, which are typically cultured on matrigel and not passaged as single cells. Furthermore, getting some stem cells to just attach and grow on chips is notoriously difficult. The team has very strong expertise in microfluidic technology but reviewers raised concern that there was not sufficient experience and expertise related to stem cell biology, which was reflected in the vagueness of the study descriptions in the biology parts of the application. Overall the technology and engineering generated great excitement, especially the collaboration between two companies.