TY - JOUR
T1 - Induced pluripotent stem cells from GMP-grade hematopoietic progenitor cells and mononuclear myeloid cells
AU - Ohmine, Seiga
AU - Dietz, Allan B.
AU - Deeds, Michael C.
AU - Hartjes, Katherine A.
AU - Miller, David R.
AU - Thatava, Tayaramma
AU - Sakuma, Toshie
AU - Kudva, Yogish C.
AU - Ikeda, Yasuhiro
N1 - Funding Information:
We thank Ms. Ying Li and Mr. Christopher P. Kolbert (Mayo Advanced Genomics Technology Center) and Mr. Scott I. Gamb (Mayo Electron Microscopy Core Facility) for excellent technical support for transcriptome analysis and electron microscopy. This work was supported by Mayo Foundation, Eisenberg Stem Cell Trust, Minnesota Partnership Grant, Marriott Individualized Medicine Award, Bernard and Edith Waterman Pilot Grants (to YI), Marriott Specialized Workforce Development Award in Individualized Medicine (TT), and NIH Grant DK 085516 (YK).
PY - 2011
Y1 - 2011
N2 - Introduction. The induced pluripotent stem cell (iPSC) technology allows generation of patient-specific pluripotent stem cells, thereby providing a novel cell-therapy platform for severe degenerative diseases. One of the key issues for clinical-grade iPSC derivation is the accessibility of donor cells used for reprogramming. Methods. We examined the feasibility of reprogramming mobilized GMP-grade hematopoietic progenitor cells (HPCs) and peripheral blood mononuclear cells (PBMCs) and tested the pluripotency of derived iPS clones. Results: Ectopic expression of OCT4, SOX2, KLF4, and c-MYC in HPCs and PBMCs resulted in rapid iPSC derivation. Long-term time-lapse imaging revealed efficient iPSC growth under serum- and feeder-free conditions with frequent mitotic events. HPC- and PBMC-derived iPS cells expressed pluripotency-associated markers, including SSEA-4, TRA-1-60, and NANOG. The global gene-expression profiles demonstrated the induction of endogenous pluripotent genes, such as LIN28, TERT, DPPA4, and PODXL, in derived iPSCs. iPSC clones from blood and other cell sources showed similar ultrastructural morphologies and genome-wide gene-expression profiles. On spontaneous and guided differentiation, HPC- and PBMC-derived iPSCs were differentiated into cells of three germ layers, including insulin-producing cells through endodermal lineage, verifying the pluripotency of the blood-derived iPSC clones. Conclusions: Because the use of blood cells allows minimally invasive tissue procurement under GMP conditions and rapid cellular reprogramming, mobilized HPCs and unmobilized PBMCs would be ideal somatic cell sources for clinical-grade iPSC derivation, especially from diabetes patients complicated by slow-healing wounds.
AB - Introduction. The induced pluripotent stem cell (iPSC) technology allows generation of patient-specific pluripotent stem cells, thereby providing a novel cell-therapy platform for severe degenerative diseases. One of the key issues for clinical-grade iPSC derivation is the accessibility of donor cells used for reprogramming. Methods. We examined the feasibility of reprogramming mobilized GMP-grade hematopoietic progenitor cells (HPCs) and peripheral blood mononuclear cells (PBMCs) and tested the pluripotency of derived iPS clones. Results: Ectopic expression of OCT4, SOX2, KLF4, and c-MYC in HPCs and PBMCs resulted in rapid iPSC derivation. Long-term time-lapse imaging revealed efficient iPSC growth under serum- and feeder-free conditions with frequent mitotic events. HPC- and PBMC-derived iPS cells expressed pluripotency-associated markers, including SSEA-4, TRA-1-60, and NANOG. The global gene-expression profiles demonstrated the induction of endogenous pluripotent genes, such as LIN28, TERT, DPPA4, and PODXL, in derived iPSCs. iPSC clones from blood and other cell sources showed similar ultrastructural morphologies and genome-wide gene-expression profiles. On spontaneous and guided differentiation, HPC- and PBMC-derived iPSCs were differentiated into cells of three germ layers, including insulin-producing cells through endodermal lineage, verifying the pluripotency of the blood-derived iPSC clones. Conclusions: Because the use of blood cells allows minimally invasive tissue procurement under GMP conditions and rapid cellular reprogramming, mobilized HPCs and unmobilized PBMCs would be ideal somatic cell sources for clinical-grade iPSC derivation, especially from diabetes patients complicated by slow-healing wounds.
UR - http://www.scopus.com/inward/record.url?scp=83455186619&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=83455186619&partnerID=8YFLogxK
U2 - 10.1186/scrt87
DO - 10.1186/scrt87
M3 - Article
C2 - 22088171
AN - SCOPUS:83455186619
SN - 1757-6512
VL - 2
JO - Stem Cell Research and Therapy
JF - Stem Cell Research and Therapy
IS - 6
M1 - 46
ER -