SBIR-STTR Award

The overall aim of this research program is to develop a single-cell bioreactor system that will automate the technology for culturing single cells and allow for the rapid selection and optimization of media and growth factors for stem cell growth. This system, which will allow cells to be grown in a sterile environment in which temperature, pH, and atmosphere (e.g., humidity and pO2) are regulated, will monitor continuously in situ the behavior of individual cells (e.g., division, differentiation, and motility) in multiple wells in a 96-well plate using a computer-controlled microscope imaging system with pattern recognition software to drive a robotic pipetting assembly to automatically change media in individual wells to respond to changes in cell number (growth) and phenotype (differentiation). We believe that this new technology will: 1) allow the reproducible growth of pluripotent stem cells; 2) allow automation of the study of cell growth, including the development of optimal substrates and growth factors for cell proliferation and differentiation, in a way that is superior in speed and efficiency compared to existing manual techniques; and 3) have widespread applications to many problems in cell biology, toxicology and tissue engineering. PROPOSED COMMERCIAL APPLICATION: The automated bioreactor's first commercial application will be to grow large populations of stem cells, but we believe that the bioreactor can also be used to determine the optimum growth media for many different types of cells, including neurons, myoblasts, endothelial cells and embryonic stem cells.

Thesaurus Terms:
biomedical automation, biomedical equipment development, bioreactor, cell proliferation, hematopoietic stem cell, mass tissue /cell culture CD34 molecule, computer program /software, growth factor, growth media, method development, umbilical cord charge coupled device camera, flow cytometry, human tissue, optics, phase contrast microscopyNational Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)

This is a revised SBIR Phase II application. The proposal seeks to develop an automated cell culture and analysis system for single cells and apply this system to study of the growth of human hematopoietic stem cells. The approach is based on the hypothesis that an automated single cell bioreactor can be designed and constructed and that this system will provide a tool to advance the basic scientists' ability to dissect the basic biology of cell division and differentiation and to discover how to expand human hematopoietic stem cells in vitro. The system to be developed is a robotic system that intends to automate the processes of: 1) of cell culture and provide information at the level of individual cells; 2) screening of the effects of biological or chemical moieties on tissue-specific cells; and 3) manipulating the fluid microenvironment of cells based upon real-time identification of changes in cell state (e.g., cell division, cell phenotype, apoptosis, etc.). In this Phase II project, the performance and image processing capability of the existing system will be improved and validated; once these tasks are completed, the system's capabilities will be expanded in terms of throughput and the quality and quantity of information obtained. Applications and testing protocols developed in the Phase II program will be directed at the system's use to understand the cell biology of hematopoietic stem cells although the system ultimately would have a number of biological and drug discovery applications.