Reprogramming T cell lymphocytes to induced pluripotent stem cells
2012-12
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Reprogramming T cell lymphocytes to induced pluripotent stem cells
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2012-12
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Abstract
The discovery of induced pluripotent stem cells (iPSC) provided a novel technology for
the study of development and pharmacology and complement embryonic stem cells (ES)
for cell therapy applications. Though iPSC are derived from adult tissue they are
comparable to ES cells in their behavior; multi-lineage differentiation and self-renewal.
This makes iPSC research appealing because they can be studied in great detail and
expanded in culture broadly. Fibroblasts were the first cell type reprogrammed to an iPSC
using a retrovirus vector, since then alternative cell types including lymphocytes have
been used to generate iPSC. Different types of vectors have also been developed to
enhance iPSC formation and quality. However, specific T lymphocyte subsets have not
been shown to reprogram to a pluripotent state to date. Here, we proposed to derive iPSC
from peripheral blood effector and central memory T cells, reasoning that the resultant
iPSC will maintain the epigenetic memory of a T lymphocyte, including the T cell
receptor (TCR) gene rearrangement. This epigenetic memory will enable the
differentiation and expansion of T cell iPSC into professional T cells containing a
specific TCR. These could then be used for cell therapy to target specific antigens, as well as to improve culture techniques to expand T cells in vitro. We studied different
gene delivery methods to derive iPSC from different types of T lymphocytes. We
assessed the viability of viral transduction using flow cytometry to detect green
fluorescent marker contained in the viral construct and quantitative real time polymerase
chain reaction (qRT-PCR) to detect Oct4, Klf4, Sox2, and c-Myc gene expression. Our
results demonstrate that the Sendai virus construct is the most feasible platform to
reprogram T lymphocytes. We anticipate that this platform will provide an efficient and
safe approach to derive iPSC from different T cell subsets, including memory T cells.
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University of Minnesota M.S. thesis. December 2012. Major: Stem cell biology. Advisor: Jakub Tolar. 1 computer file (PDF); vi, 56 pages.
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Bared, Kalia. (2012). Reprogramming T cell lymphocytes to induced pluripotent stem cells. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/143800.
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