Investigating the effect of dynamic culture and lactate on colony formation of human induced pluripotent stem cells (hiPSCs)

Date
2023-09-20
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Abstract
Research into stem cell treatments has led to the ability to reprogram somatic cells in the body, like skin cells and bloods cells, to behave like stem cells, and have pluripotent characteristics, ability to self-renew and develop into almost any cell type or tissue. These reprogrammed cells, referred to as human induced pluripotent stem cells (hiPSCs), have broad applications in cell therapy, drug screening, and disease modelling. Unlike traditional pluripotent cells, these cells are more accessible, avoid ethical concerns, and can be used for autologous therapies. Various methods of reprogramming somatic cells are available but come with challenges of virus integration into the genome, low reprogramming efficiency, and long transition from cultivation to large-scale manufacturing. The proposed research addresses this by combining two innovative approaches for improved somatic cell reprogramming: 1. transient exposure to lactate, and 2. dynamic culture environment. The generation of hiPSCs is highly dependent on a metabolic shift from oxidative phosphorylation (OXPHOS) to glycolysis and lactate has shown to facilitate this metabolic shift thereby potentially increasing reprogramming efficiency. The dynamic environment also plays a role in reprogramming and the subsequent physiology which allows for great cell expansion, heterogeneity and scale-up. This study first outlines the isolated effects of a dynamic system and transient lactate exposure on the reprogramming process as well as the optimization of these conditions. The two approaches were then integrated with findings suggesting that the independent conditions outperformed the tandem. Further phenotype and functional testing post reprogramming is required, however the examined conditions show promise in increasing reprogramming efficiency, reducing time of reprogramming process and improving translation to a bioprocess.
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Keywords
Stem cells, Pluripotent, Dynamic culture, Bioprocess, Lactate
Citation
Malinovska, J. (2023). Investigating the effect of dynamic culture and lactate on colony formation of human induced pluripotent stem cells (hiPSCs) (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.