Gillian Primavera
Pronouns: She/Her
Research Mentor(s): Seth Teague
Co-Presenter:
Research Mentor School/College/Department: Biomedical Engineering / Engineering
Presentation Date: April 20
Presentation Type: Poster
Session: Session 6 – 4:40pm – 5:30 pm
Room: League Ballroom
Authors:
Presenter: 44
Abstract
There are current limitations in stem cell technologies due to the variability of cell fates that arise during directed differentiation due to secondary signaling. The motivation behind the project I am involved in was to determine how heterogeneity within signaling of stem cells is linked to different observed cell fates. This project focused primarily on activating one developmental pathway, the BMP pathway, while mitigating the effect of others such as WNT and NODAL. We aimed to determine whether the amplitude of BMP signaling or the duration of signaling is the most important factor in determining cell fate in a simple binary decision between differentiating to an amnion-like fate or remaining pluripotent. To do this, we measured the cellular localization of fluorescent fusions of the BMP signal transduction proteins SMAD1 and SMAD4 proteins and we were able to validate these fusions as a reliable source for quantifying BMP response. Live imaging was conducted to track signaling reporters in single cells throughout differentiation followed by fixed immunofluorescence imaging to determine levels of cell fate markers in the same cells. We identified pluripotent cells as SOX2+/NANOG+/ISL1- and amnion-like cells as SOX2-/NANOG-/ISL1+. Relative levels of these markers were determined and cells were then divided into amnion or pluripotent based on the log expression ratio of ISL1 to NANOG. We gathered significant data that shows that signaling is approximately binary and the duration of high signaling is more important than its absolute amplitude.
Engineering
