Going Back and Forth: Episomal Vector Reprogramming of Peripheral Blood Mononuclear Cells to Induced Pluripotent Stem Cells and Subsequent Differentiation into Cardiomyocytes and Neuron-Astrocyte Co-cultures

Abstract

Human induced pluripotent stem cells (iPSCs) can capture the diversity in the general human population as well as provide deeper insight in cellular mechanisms. This makes them suitable to study both fundamental and applied research subjects, such as disease modeling, gene-environment interactions, personalized medicine, and chemical toxicity. In an independent laboratory, we were able to generate iPSCs originating from human peripheral blood mononuclear cells according to a modified version of a temporal episomal vector (EV)-based induction method. The iPSCs could subsequently be differentiated into two different lineages: mesoderm-derived cardiomyocytes and ectoderm-derived neuron-astrocyte co-cultures. It was shown that the neuron-astrocyte culture developed a mature phenotype within the course of five weeks and depending on the medium composition, network formation and neuron-astrocyte cell ratios could be modified. Although previously it has been described that iPSCs generated with this EV-based induction protocol could differentiate to mesenchymal stem cells, hepatocytes, cardiomyocytes, and basic neuronal cultures, we now demonstrate differentiation into a culture containing both neurons and astrocytes.

Somatic Cell Reprogramming for Nervous System Diseases: Techniques, Mechanisms, Potential Applications, and Challenges

“…For example, induced pluripotent stem cells (iPSCs) such as peripheral blood mononuclear cells display pluripotency after transdifferentiation; they can be converted into motor neurons using differentiation medium containing brain-derived neurotrophic factor (BDNF) or glial cell line-derived neurotrophic factor (GDNF) [ 30 , 54 ]. The resulting neurons express mature neuronal markers such as Tuj1, Map2, and synaptophysin, and they show spontaneous action potentials in patch clamp assays [ 14 , 15 , 16 , 17 ].…”

“…This achievement was considered a revolutionary advancement in cell biology and medicine and laid the foundation for subsequent research. In subsequent studies, scientists discovered that it was possible to obtain mature neurons by converting somatic cells into stem cells and then differentiating them into neural precursor cells with a dozen transcription factors [ 14 , 15 , 16 , 17 ].…”