Towards biology-driven strategies for stem cell therapy applications

Abstract

Towards the second goal, we demonstrate that MSCs can be selectively enriched using mitochondrial and plasma membrane potential indicators. Electrically enriched MSCs are phenotypically distinct cells with distinct immunoregulatory capacities. In addition, we demonstrate that MSCs possess bioelectrical plasticity and introduce novel assay that can be utilized to assess bioelectrical phenotype of MSCs.

Towards the first goal, we utilized a combinatorial approach of doxycycline-regulated expression of transcription factors FOXD3 and encapsulation of ESCs into collagen-functionalized hydrogel. We demonstrated that this approach leads to decreased pluripotency, increased specificity and enhanced osteogenic differentiation of ESCs. We demonstrate that the ability to tune the expression of genes responsible for pluripotency or differentiation commitment may provide a novel tool for developing safer and more efficient cell therapy products.

Stem cell therapy products have penetrated the medical market offering the treatment options for a variety of degenerative and inflammatory disorders, such as osteoporosis, osteoarthritis, macular degeneration and others. The advancement of stem cell therapies into clinical trials and practice currently outpaces the research necessary to ensure the consistent safety and efficiency of the final cell therapy products. Due to the plastic nature of stem cells, it is important to develop strategies to clinically administer stem cell products with precisely defined characteristics and in their most functional state. Those strategies must be tailored to the stem cell type to be used (e.g. mesenchymal stem cells, embryonic stem cell line, induced pluripotent stem cells) and application. The goal of this work is two-fold: 1) to develop a proof-of-concept strategy to treat bone defects and fractures by increasing specificity of osteogenic differentiation of embryonic stem cell lines (ESCs) and (2) to explore the potential of targeting bioelectrical properties of mesenchymal stem cells (MSCs) to enrich for stem cells with therapeutically desired phenotypes.