This SBIR Phase II research is focused on methods to utilize blood precursor cells derived from human embryonic stem (HES) cells. The project uses a new defined differentiation system which allows automation and scale-up production of this important cells. There is a significant demand for these cells from research and drug discovery. Increased availability and batch-to-batch reproducibility of HES cell-derived blood cells, resulting from the defined genetic background of the starting material and this standardized, automated culture system, make this technology invaluable model systems for basic research and drug development. Based on the automated pilot system for handling and scale-up production of HES cells developed in phase I of this SBIR project we will transfer our current culturing protocols into robust automated production procedures to provide a reproducible quality of CD34 positive cells
The broader impacts of this research will be improving the process of drug discovery and development and in the long term by providing revolutionary new applications for medical treatment to improve public health. Nearly 98% of a multi-million dollar stem cell market is currently consumed by blood and immune system treatments. We anticipate that the proposed research will lead to the faster integration of HES cell biology into biomedical research. It will help to provide a variety of other blood cell types in quantities required for basic research, drug development, high throughput screening, biochemical characterization and potential medical treatment of blood related disease.
Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) have the ability to proliferate for extended periods of time in an undifferentiated state and are capable of forming all cell types present in the human body. Stem cells require a plethora of different environments for proliferation and differentiation. Scaling up pluripotent stem cells and directing differentiation to different cell types is technically challenging, labor-intensive and displays inherent process variability. From an economic and regulatory process-control perspective, manual processing of stem cells cannot be a long-term solution to meet the needs of the market. The present project was focused on automating the growth, maintenance and expansion of hESCs and hiPSCs using the Tecan Cellerity system (an industrially relevant robotic platform). By industrializing the production of stem cells and their derivative cell types under good manufacturing practice (GMP), we are able to maximize the broad impact on potential regenerative medicine applications. Utilizing the unique capabilities of robotic cell culture system, we successfully generated blood precursor cells from hESCs and hiPSCs that are often found only in sparse numbers in post-natal tissues. We optimized the generation of blood precursor cells from hESCs and hiPSCs using a defined feeder-free protocol to facilitate automated screening and scalability to meet the demands of the market. The project involved significant optimization in robotic handling maneuvers with stem cells. Tecan supported the team with training sessions. Automation of hESCs and hiPSCs did not alter the genetic stability or the potential to form blood precursor cells. The precursor cells were capable of generating several defined cell types found circulating in blood. We also screened over 2000 small molecules to improve the efficiency and quality of blood precursor stem cells utilizing the automated Tecan Cellerity platform. We successfully identified unique molecules that augmented the generation of blood forming precursors from hESCs. This project demonstrated the capability of the Tecan Cellerity to maintain, differentiate and perform high throughput screening using hESCs and hiPSCs. The team included participation of instrumentation specialists, scientists and REUs. The REUs gained an educational insight to the field of automation and the potential of stem cells. The project enhanced our scientific understanding, enabling us to control the growth, maintenance and differentiation of stem cells into blood precursor cells as well as specialized cell types with increased yield and precision. The last 6 months of the project was focused on testing a panel of hiPSCs to generate blood precursor cells and a pilot collaboration with BellBrook Labs (located in Madison, WI) to test the commercialization of hiPSCs derived cell types in cellular assays kits.
