Real Time Gene Expression Analysis And Gene Based Sorting In Live Cells

Investigator: Hetal Desai

Human adipose-derived stem cells (ASCs) are attractive cells for regenerative therapies because of their lack of immunogenicity and applicability to musculoskeletal tissues. A drawback of ASCs is their heterogeneous response to biochemical stimuli during induction. Non-differentiating cells within freshly isolated ASC populations potentially interfere with the therapeutic capacity of cells that can differentiate. However, other cellular subpopulations may be predisposed to particular lineages. Isolating these cells using gene expression markers can maximize cell yield during sorting while also improving therapeutic capacity by collecting cells primed for regenerating specific tissues.

The objective of this project is to determine whether exposure to osteogenic, adipogenic, and chondrogenic induction factors causes highly responsive ASC subpopulations to express characteristic early marker genes, which act as a biomarkers for cells capable of undergoing osteogenesis, adipogenesis, and chondrogenesis. The process has been tested with osteogenesis; ALPL+ ASCs were sorted by fluorescence activated cell sorting (FACS) and osteogenically differentiated. Positively expressing cells displayed significantly large increases in calcified matrix deposition over unsorted and non-expressing cells, suggesting greater therapeutic potential for bone.