CBE Research Report Spring 2016 | Page 9

Graduate Student Oral Presentation 2 Metabolic Reprograming during Non-adherent Culture Selects Primitive Human Mesenchymal Stem Cells Yijun Liu1, Ang-Chen Tsai1, Teng Ma1 1 Chemical and Biomedical, Florida State University Abstract Removal of human mesenchymal stem cells (hMSCs) from in vivo niche for in vitro bioprocessing in an artificial culture environment induces senescence that reduce hMSC potency and impede translation of hMSC to clinical application1. Recent studies on the non-adherent culture of anchorage-dependent cells suggest the strong correlation between self-assembled aggregates and cellular ‘stemmess’, and indeed aggregates of hMSCs exhibit enhanced pluripotent stem cells genes expression and primitive stem cells functions. Although it has been suggested that in vitro aggregation mimics mesenchymal condensation and recapitulates the cellular events of in vivo microenvironment, the molecular link between aggregation and hMSCs stemness is still undefined. In the present study, we test the hypotheses that hMSC aggregation on non-adherent substratum initiates mitochondrial remodeling and metabolism reconfiguration, resulting in a metabolic profile that resembles primitive stem cells. Our results show that aggregation of hMSCs influences mitochondrial dynamics and results in a reduction in mitochondrial function. Adaptive changes as a result of mitochondrial dysfunction includes increased glycolytic and anaplerotic flux, which fuel the self-digestion program named autophagy. Importantly, we identify that alteration of mitochondrial respiration chain is responsible for the enhanced expression of stem cell genes because treatment of respiration chain targeted small molecules partially recapitulates stemness enhancement event in planar culture. Our finding demonstrates that mitochondria play significant role in stem cell pluripotency by its retrograde signaling with stem cell gene expression program (Fig.1). Reference 2. Ma T, Tsai A, Liu Y. 2015. Biomanufacturing of Human Mesenchymal Stem Cells in Cell Therapy: Influence of Microenvironment on Scalable Expansion in Bioreactors. Biochemical Engineering Journal. 108: 44. Fig.1 Mitochondrial retrograde signaling in hMSCs aggregates. 8|Page