Open AccessThe RUNX1 +24 Enhancer and P1 Promoter Identify a Unique Subpopulation of Hematopoietic Progenitor Cells Derived from Human Pluripotent Stem Cells
Author(s) -
Ferrell Patrick I.,
Xi Jiafei,
Ma Chao,
Adlakha Mitali,
Kaufman Dan S.
Publication year - 2015
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.1940
Subject(s) - biology , induced pluripotent stem cell , stem cell , haematopoiesis , microbiology and biotechnology , embryonic stem cell , progenitor cell , cd34 , kosr , population , induced stem cells , adult stem cell , embryoid body , genetics , gene , demography , sociology
Derivation of hematopoietic stem cells (HSCs) from human pluripotent stem cells remains a key goal for the fields of developmental biology and regenerative medicine. Here, we use a novel genetic reporter system to prospectively identify and isolate early hematopoietic cells derived from human embryonic stem cells (hESCs) and human induced pluripotent cells (iPSCs). Cloning the human RUNX1c P1 promoter and +24 enhancer to drive expression of tdTomato (tdTom) in hESCs and iPSCs, we demonstrate that tdTom expression faithfully enriches for RUNX1c ‐expressing hematopoietic progenitor cells. Time‐lapse microscopy demonstrated the tdTom + hematopoietic cells to emerge from adherent cells. Furthermore, inhibition of primitive hematopoiesis by blocking Activin/Nodal signaling promoted the expansion and/or survival of the tdTom + population. Notably, RUNX1c/tdTom + cells represent only a limited subpopulation of the CD34 + CD45 + and CD34 + CD43 + cells with a unique genetic signature. Using gene array analysis, we find significantly lower expression of Let‐7 and mir181a microRNAs in the RUNX1c/tdTom + cell population. These phenotypic and genetic analyses comparing the RUNX1c/tdTom + population to CD34 + CD45 + umbilical cord blood and fetal liver demonstrate several key differences that likely impact the development of HSCs capable of long‐term multilineage engraftment from hESCs and iPSCs. S tem C ells 2015;33:1130–1141