[생명공학연구소] Dynamics of repopulation by long-term non-human primate HPSC revealed by a sensitive, quantitative, and high-t
2010.09.08비서실3379
- 일시 : 2010.09.09
◈ 주제 : Dynamics of repopulation by long-term non-human primate HPSC revealed by a sensitive, quantitative, and high-throughput clonal tracking assay
◈ 연사 : 김상구 박사
◈ 소속 : UCLA, AIDS Institute
◈ 일시 : 2010년 9월 9일(목) 오후 5:00
◈ 장소 : 생명공학관 333호
<초록>
Insight into the mechanism of long-term hematopoietic progenitor and stem cell (HPSC) repopulation following transplant is critical to the advancement of HPSC therapy. We developed a highly sensitive and quantitative method using pyrosequencing technology to measure the relative frequencies of individual clones in a complex polyclonal circulating cell population. We applied this assay to study HPSC repopulation in a rhesus macaque transplanted for 10 years with autologous CD34+ cells transduced with self-inactivating lentiviral vectors. We estimated that over a thousand stem cell clones repopulate long-term. These clones repopulate with heterogeneous kinetics that can be grouped into discrete patterns of emergence and decline. While frequencies of individual clones varied over time, the majority (80%) of clones, detected as early as 5 and 7 months post-transplant, persisted long-term (4 to 10 years post-transplant). Sequential expansions of some clones were balanced by reductions of other clones, which maintained homeostasis at a relatively constant level of overall repopulation from 10 months to 10 years post-transplant. A small number of clones (25 clones) predominately contributed to long-term hematopoiesis, accounting for over 50% of total repopulation. This novel assay provides the first in-depth kinetic study of long-term stem cell clonal repopulation in primates and can be a useful clinical tool for assessment of repopulation and safety in humans
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