VCOM Research Day Program Book 2023

Faculty Research Biomedical

03 Effects of Meis1 Suppression on Mouse Embryonic Stem Cell Proliferation and Viability

Ahmed Elywa; Ranya Ridha; Sondos Elnady; Shreya Raj; Suzanne Tunder; Jia-Qiang He Corresponding author: elywa2022@vt.edu

Department of Biomedical Sciences and Pathobiology, College of Veterinary Medicine

Cardiovascular diseases (CVDs) remain the leading cause of death in the United States of America. The major impediment to get full recovery from ischemic heart diseases and myocardial infarctions is the inability of adult mammalian cardiac tissue to regenerate after damage, which leads to decreased cardiac contractility and subsequent heart failure. The aim of this study is to investigate the role of myeloid ecotropic viral integration site 1 (Meis1) gene in regulating proliferation and viability of mouse embryonic stem cells (mESCs), which can be later differentiated into functional cardiomyocytes and used as a promising therapeutic modality for future

treatment of cardiac diseases. Based on published data and our preliminary results, we hypothesize that Meis1 suppression can negatively affect mESCs proliferation but positively promote cardiomyocytes differentiation in vitro. As a result, mESCs were cultured and maintained aseptically in culture plates using DMEM/High glucose culture medium supplemented with growth factors and antibiotics and kept in incubator at 37oC and 5 % CO2. The cultured cells were then used to extract mRNA to quantify Meis1 gene expression level using real time PCR technique. Total protein was also extracted from cultured mESCs for detection of Meis1 protein via

Western Blot technique. In addition, the cultured cells were examined for its viability using Calcein AM/EthD-1/Hoechst combined fluorescent staining, while the cell proliferation was assessed using EdU fluorescent staining. The ongoing experiments are checking whether inhibition of Meis1 gene modulates the cell viability and proliferation utilizing the same methods described above.

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