EAGER: Biomanufacturing: Gene expression-based standardization of stem cells

Brown University, Providence RI

Investigators

Abstract

PI: Darling, Eric M. Proposal Number: 1547819 Cell-based biomanufacturing is highly dependent on its starting material: the cell. Using poorly defined, mixed populations of cells will result in unsatisfactory outcomes, regardless of the downstream technologies. Purification or enrichment of high-value material, like adult stem cells, is necessary for a broad range of existing and future therapeutic strategies. By better controlling the composition of the source population of cells, the performance of regenerative therapies and products is hypothesized to be more reproducible, making biomanufacturing approaches more feasible than they are today. Current enrichment techniques, which depend on the existence of unique combinations of cell surface proteins, are limited by very low cell yields and significant susceptibility to donor variability. In this work, the investigators propose to develop and evaluate new procedures that enable the production of stem cell populations with consistent regenerative characteristics, independent of donor. The overall goal of this proposal is to rapidly isolate primary, mesenchymal stem cells and enrich for subpopulations capable of regeneration using an innovative cell separation device in conjunction with gene expression-based enrichment. This project will be a collaboration among researchers at Brown University, Rhode Island Hospital, and Endocellutions, Inc. To achieve this goal, a live-cell molecular beacon will be designed that fluorescently identifies cells capable of adipogenic gene expression. Human fat tissue will be obtained using the Adi-Cellutions device, and cells within the solid/liquid fractions of the isolate will be examined for their regenerative characteristics following gene expression-based sorting. Two specific aims are included in this proposal: 1) Investigate how gene expression-based cell sorting parameters influence high-yield acquisition of adipogenic cells, and 2) Establish a set of processing parameters capable of generating consistent regenerative characteristics in isolated cell populations, independent of donor. Gene expression-based cell enrichment is a potential, paradigm-shifting technique, vastly expanding the potential targets researchers can investigate beyond current, cell surface marker approaches. More practically, the methodologies generated from this project will provide a roadmap for other investigators to obtain more consistent populations of cells for use in basic science experiments as well as clinical procedures. The proposed academic-industry collaboration will result in contributions to the fields of biomedical engineering, cell and molecular biology, and clinical medicine. Undergraduate, graduate, and medical students will participate throughout all stages of the proposed project, and experimental findings will be distributed through scientific communications and educational venues at Brown University and partnering hospitals.