Researchers in a tissue engineering lab have developed a new…

Researchers in a tissue engineering lab have developed a new strategy to reprogram adult cells and generate IPSCs using a novel viral vector that can efficiently deliver plasmid into the adult cells. In order to track in real time the reprogramming efficiency of their method they use a cell line engineered with a GFP-reporter on the Oct4 gene (GFP: Green fluorescent protein). In the figure presented below, the researchers induced the reprogramming process at day #7 and analyzed the reprogramming efficiency by comparing the total cell confluence (GFP-labelled and non-labelled cells) with the GFP expressing cells confluence (GFP-labelled cells). (In cell culture biology, confluence refers to the percentage of the surface of a culture dish that is covered by adherent cells. The more cells are growing the more confluent the culture is). Using your knowledge on the IPSCs reprogramming process and the results presented in the figure, please answer the following questions. a. What does the acronym IPSCs stand for? b. What type of cell are often use to generate IPSCs (be as specific as you can)? c. Provide two examples of Transcription factors that can be used to reprogram fibroblasts into IPSCs. d. What is Oct4 and why are the researchers monitoring Oct4 gene expression? Provide an example of other gene that the authors could have used or monitor in this study. e. Instead of viral delivery, what other method could be used to reprogram adult cells into IPSCs? f. Please describe the results in the figure below. g. Based on these results can we say that this new method is able to generate IPSCs? and if yes how would you determine the efficiency? Figure caption: Assessing Reprogramming Kinetics by Measuring OCT4-GFP Confluence. Real-time imaging of fibroblasts reprogrammed with a new viral vector. Phase contrast, green fluorescence and merged images of the same cell colonies were generated at Day 7 to 19. Scale bar corresponds to 400 μm. The graph represent the quantification of total cell confluence (phase contrast) and OCT4-GFP cell confluence (green channel).