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Thesis - Campus Access Only
Master of Science (MS)
The ability to create tissues in vitro from human embryonic stem cells (hESCs) would allow for the production of cells to address currently unmet therapeutic and pharmaceutical needs. To meet these needs, it is necessary to establish a scheme whereby a highly pure population of a given therapeutically relevant cell type (e.g., cardiomyocytes) may be efficiently generated from hESCs. Upon differentiation, hESCs yield only a small fraction of cardiomyocytes together with other miscellaneous cell types, which is unsuitable for any practical application. To overcome this limitation, we sought to identify prospective cell surface markers on mesoderm progenitors, the developmental precursor to all cardiac cell types. Utilizing a reporter hESC line in which GFP is inserted into one allele of the transcription factor, Mixl1, mesoderm cells emerging upon differentiation in vitro were identified and sorted using fluorescence- activated cell sorting (FACS). Microarray analysis of purified fractions was then used to identify two genes, CD13 and ROR2, which encode surface proteins specifically expressed in an early phase of mesoderm development. By enabling the positive selection of mesoderm precursors from differentiated hESC populations, CD13 and ROR2 should allow for the enrichment of subsequent cardiac fractions as well as for facilitating the necessary removal of contaminating cell types that plague current protocols.
Brady, Bevin, "CD13 and ROR2 Permit Prospective Isolation of Purified Mesoderm Progenitors from Human Embyonic Stem Cell Differentiation" (2013). Master's Theses. 4378.