An Integrated Microfluidic Platform for Screening hESC Culture Conditions

Funding Type: 
SEED Grant
Grant Number: 
RS1-00314
Investigator: 
ICOC Funds Committed: 
$0
oldStatus: 
Closed
Public Abstract: 
Microfluidic systems handle and manipulate tiny amounts of fluids at volumes thousands times smaller than a tear drop. The goals of our proposal are (i) to design, fabricate and test a microfluidic technology platform (i.e., hESC-Chips) for performing human embryonic stem cell (hESC) culture and assay with significantly improved efficiency, (ii) to utilize these hESC-Chips in search of new culture conditions for hESC self renewal and (iii) to disseminate this technology for routine use in other hESC laboratories. Compared to those macroscopic setting employed for the conventional hESC research, the advantages of the proposed hESC-Chips are low sample/reagent consumption, fast processing, precise control over physical/chemical environments and automated operation. As the proposed project unfolds, we anticipate having contributions on the following three aspects: First, the successful demonstration of the proposed hESC-Chips will provide a powerful technology for contemporary hESC research. In our design, a hESC-Chip will contain 100 culture chambers for a simultaneous examination of 100 hESC experiments. Since the sizes of these hESC culture chambers are very small, the consumptions of hESC samples and the associated reagents will be significantly reduced (4 to 5 orders of magnitude lower than the conventional setting). In addition, due to the use of PC-operated interface, critical parameters for hESC experiments can be monitored and controlled in the hESC-Chips with superior precision, which is unattainable using the conventional macroscopic setting. Second, the hESC-Chips promise to accelerate the screening process in search of animal product-free culture conditions for hESC self renewal and differentiation. Generally, hESC culture requires feeders, e.g., mouse embryonic fibroblasts (mEFs) or Matrigel, and mEF-conditioned medium to maintain hESCs at the undifferentiated stage. These culture conditions lead to contamination of animal products, thus restrict the therapeutic applications of hESCs in clinic settings. Right now, it is critical to define animal product-free hESC culture conditions by screening a large number of combinations of extracellular matrices and soluble factors. Using the conventional hESC research setting this screening process is time and cost-consuming. Third, we will disseminate this microfluidic technology for routine use in other hESC laboratories. A user friendly version of hESC-Chips will be developed and then tested in the co-PI’s research group and other hESC groups at UCLA. The final versions of the chip design and control programs will be freely available for download from the PI’s research group web site (http://labs.pharmacology.ucla.edu/tsenglab/), where some existing designs and programs can be accessed.
Statement of Benefit to California: 
As the proposed project unfolds, we anticipate will benefit the State of California and it citizens on five fronts: 1. The successful demonstration of the proposed microfluidic platform (i.e., hESC-Chips) will present a new technology for far-reaching application to all types of stem cell research with significantly improved operation efficiency. In contrast, the conventional stem cell research is plagued by the use of macroscopic setting, resulting in several constraints, e.g., high sample/reagent consumption, poor precision to control the environments of hESC experiments and the lack of integrated platforms for accurate measurements. 2. Using the proposed hESC-Chips the costs for hESC research will be greatly reduced. We estimate that each hESC experiment in the hESC-Chip consumes 30 nanoliter of cell culture reagents/medium, which is 4 to 5 orders of magnitude lower than a commonly used 6-well plate (requiring 2.5 mL of reagents/medium for a single study). Many hESC studies that require large-scale hESC experiments can, therefore, be implemented using the hESC-Chips in a cost-efficient manner. 3. The proposed hESC-Chip allows greater precision of measurements. Due to the use of PC-operated interface, critical parameters for hESC experiments can be monitored and controlled in the hESC-Chips with superior precision, which is unattainable using the conventional macroscopic setting. 4. The hESC-Chips promise to accelerate the discovery of new understanding of hESC growth control and to speed up the drug screening processes for hESC therapy. In our design, a hESC-Chip will contain 100 culture chambers for a simultaneous examination of 100 hESC experiments. A custom-designed interface allows 50 hESC-Chips to function in parallel. 5. User-friendly instrumentations of the proposed technology will be created for routine use in other hESC laboratories. The final versions of the chip design and control programs will be freely available for download from the PI’s research group web site (http://labs.pharmacology.ucla.edu/tsenglab/), where some existing designs and programs can be accessed.

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