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Date of Award
5-2025
Document Type
Restricted Thesis: Campus only access
Degree Name
Master of Science in Biology
Department
Biology
First Reader/Committee Chair
Bournias-Vardiabasis, Nicole
Abstract
Human neonatal cardiovascular tissue contains a unique and varied repertoire of early-stage stem cells based on RNA sequencing and transcriptomic analysis of clonal cell subpopulations. The current study sought to determine whether early-stage stem cells with differentiation capability along the hematopoietic lineage exist transiently in cardiac tissue derived from human neonates. We hypothesized that Islet-1+ neonatal subpopulations expressing ETV2, MESP1, EOMES, and MIXL1 can differentiate into the hematopoietic lineage and exhibit a distinct stage-specific microRNA profile reflective of their multipotent capabilities.
To test this hypothesis, clonal subpopulations of human Islet-1+ stem cells isolated from discarded cardiac surgical tissue of neonates were analyzed by transcriptomics and RT-qPCR. Clones expressing pre-mesendodermal markers ETV2, MESP1, EOMES, and MIXL1 were differentiated through supplementation of growth factors BMP4, FGF2, SCF, VEGF, IL3, and FLT3 at specific intervals over ten days. The expression of hematopoietic markers CD34 and CD43 was evaluated using RT-qPCR and gel electrophoresis to assess differentiation progression. Additionally, the microRNA profiles of untreated Islet-1+ clonal subpopulations were analyzed using RNAseq-based transcriptomic data.
Results indicated a significant increase in the expression of early-stage hematopoietic markers, CD34 and CD43 after differentiation, suggesting commitment toward the hematopoietic lineage. The microRNA profile of the clonal subpopulations demonstrating differentiation along the hematopoietic lineage revealed elevated expression of hematopoiesis-related microRNAs, including miR-Let-7d, miR-10a, miR-125a, miR-125b, and miR-181a. Our results define a select group of early-stage transcripts and microRNAs which are expressed in neonatal Islet-1+ subpopulations with the ability to differentiate along the hematopoietic lineage.
Recommended Citation
Melikian, Raphaella Haroutioun, "TRANSIENT POPULATIONS OF EARLY-STAGE STEM CELLS WITH HEMATOPOIETIC LINEAGE DIFFERENTIATION CAPABILITY RESIDE IN NEONATAL CARDIOVASCULAR TISSUE" (2025). Electronic Theses, Projects, and Dissertations. 2293.
https://scholarworks.lib.csusb.edu/etd/2293