Interrogating ground state rhesus macaque and human pluripotent stem cells
| dc.contributor.advisor | Zhao, Haiqing | |
| dc.contributor.committeeMember | Horner, Robert D | |
| dc.contributor.committeeMember | Tifft, Kathryn | |
| dc.contributor.committeeMember | Zambidis, Elias T | |
| dc.creator | Pather, Sarshan Rubintheran | |
| dc.date.accessioned | 2019-04-15T03:23:40Z | |
| dc.date.created | 2015-05 | |
| dc.date.issued | 2015-04-10 | |
| dc.date.submitted | May 2015 | |
| dc.date.updated | 2019-04-15T03:23:40Z | |
| dc.description.abstract | Iterative chemical screening has resulted in optimized culture conditions to support the self-renewal of human and non-human primate pluripotent stem cells (PSCs) in a mouse embryonic stem cell-like ground state of pluripotency. While all protocols employ leukemia inhibitory factor (LIF) supplemented with MEK/ERK and GSK3-β inhibitors (LIF 2i), many indispensably require additional anti-apoptotic small molecules and primed growth factors. The finding of a minimal cocktail of LIF 2i plus axin-stabilizing XAV939 (termed LIF 3i) that efficiently reverts select myeloid progenitor hiPSC lines to a ground state advances the possibility that among heterogeneous PSCs, many lines are epigenetically more amenable to naïve reversion than others. Here, I report characterization of LIF 3i-permissive human naïve PSCs by examining transcriptome microarray data and OCT3/4 enhancer DNA methylation profiles. Additionally, I report an optimized feeder-dependent culture system to support naïve-like self-renewal in non-permissive rhesus macaque embryonic stem cells (rESCs) derived from in vitro fertilization (IVF), somatic cell nuclear transfer (SCNT), and parthenogenesis. Use of a potent VEGF receptor inhibitor, sunitinib malate, together with LIF 3i or LIF 2i, promoted naïve-like ground state self-renewal in rESCs. Efficient derivation and careful interrogation of human and non-human primate naïve PSCs before in vitro differentiation, plasmid-based gene targeting, or blastocyst complementation experiments is a necessary process to advance naïve pluripotency in stem cell biology and regenerative medicine. | |
| dc.format.mimetype | application/pdf | |
| dc.identifier.uri | http://jhir.library.jhu.edu/handle/1774.2/60373 | |
| dc.language.iso | en_US | |
| dc.publisher | Johns Hopkins University | |
| dc.publisher.country | USA | |
| dc.subject | Ground state pluripotency | |
| dc.subject | primed pluripotency | |
| dc.subject | rhesus macaque embryonic stem cells | |
| dc.subject | human embryonic stem cells | |
| dc.subject | human induced pluripotent stem cells | |
| dc.title | Interrogating ground state rhesus macaque and human pluripotent stem cells | |
| dc.type | Thesis | |
| dc.type.material | text | |
| local.embargo.lift | 2019-05-01 | |
| local.embargo.terms | 2019-05-01 | |
| thesis.degree.department | Biology | |
| thesis.degree.discipline | Cell Biology | |
| thesis.degree.grantor | Johns Hopkins University | |
| thesis.degree.grantor | Krieger School of Arts and Sciences | |
| thesis.degree.level | Masters | |
| thesis.degree.name | M.S. |