| dc.description.abstract |
<p>The differentiation of human induced pluripotent stem cells (hiPSCs) to prescribed
cell fates enables the engineering of patient-specific tissue types, such as hyaline
cartilage, for applications in regenerative medicine, disease modeling, and drug screening.
In many cases, however, these differentiation approaches are poorly controlled and
generate heterogeneous cell populations. In this dissertation, we demonstrate robust
cartilaginous matrix production in multiple hiPSC lines using a robust and reproducible
differentiation protocol. To purify chondroprogenitors produced by this protocol,
we engineered a COL2A1-GFP knock-in reporter hiPSC line by CRISPR-Cas9 genome editing.
Purified chondroprogenitors demonstrated an improved chondrogenic capacity compared
to unselected populations, improved matrix homogeneity, and reduced variability between
tissues. We next demonstrated the ability of the system to serve as a high-throughput
system for arthritis disease modeling using cytokine stimuli. Finally, we used this
platform to screen for transcription factors whose activation might be involved in
chondrogenic lineage specification of hiPSCs. Taken together, these studies describe
the generation of a high-throughput system for chondrogenesis and its application
for screens and arthritis disease modeling. Future applications of this platform may
be useful for identifying pathways regulating cartilage regeneration and novel therapeutics
for arthritis.</p>
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