TMH477357_sm_Suppl._Material.pdf (551.6 kB)
Supplementary Material for: Emergence of CD43-Expressing Hematopoietic Progenitors from Human Induced Pluripotent Stem Cells
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posted on 2017-05-23, 06:34 authored by Kessel K.U., Bluemke A., Schöler H.R., Zaehres H., Schlenke P., Dorn I.Background: The ex vivo generation of human hematopoietic
stem cells (HSCs) with long-term repopulating
capacity and multi-lineage differentiation potential represents
the holy grail of hematopoiesis research. In principle,
human induced pluripotent stem cells (hiPSCs) provide
the tool for both studying molecular mechanisms of
hematopoietic development and the ex vivo production
of ‘true’ HSCs for transplantation purposes and lineagespecific
cells, e.g. red blood cells, for transfusion purposes.
CD43-expressing cells have been reported as the
first hematopoietic cells during development, but
whether or not these possess multilineage differentiation
and long-term engraftment potential is incompletely understood.
Methods: We performed ex vivo generation of
hematopoietic stem cells from hiPSCs using an embryoid
body(EB)-based, xeno-product-free differentiation protocol.
We investigated the multilineage differentiation potential
of different FACS-sorted CD43-expressing cell subsets
by colony-forming assays in semisolid media. Further,
erythroid differentiation was investigated in more
detail using established protocols. Results: By using
CD43 as a marker, we are able to measure hematopoietic
induction efficiency during hiPSC-derived EB differentiation.
Further, we determined CD43+ cells as the cell population
of origin for in vitro erythropoiesis. CD43hi CD45+
cells represent hematopoietic stem and progenitor cells
with multilineage differentiation potential with a bias towards
the erythroid lineage.