TY - JOUR
T1 - Defined and Scalable Differentiation of Human Oligodendrocyte Precursors from Pluripotent Stem Cells in a 3D Culture System
AU - Rodrigues, Gonçalo M.C.
AU - Gaj, Thomas
AU - Adil, Maroof M.
AU - Wahba, Joyce
AU - Rao, Antara T.
AU - Lorbeer, Franziska K.
AU - Kulkarni, Rishi U.
AU - Diogo, Maria Margarida
AU - Cabral, Joaquim M.S.
AU - Miller, Evan W.
AU - Hockemeyer, Dirk
AU - Schaffer, David V.
N1 - Publisher Copyright:
© 2017 The Author(s)
PY - 2017/6/6
Y1 - 2017/6/6
N2 - Oligodendrocyte precursor cells (OPCs) offer considerable potential for the treatment of demyelinating diseases and injuries of the CNS. However, generating large quantities of high-quality OPCs remains a substantial challenge that impedes their therapeutic application. Here, we show that OPCs can be generated from human pluripotent stem cells (hPSCs) in a three-dimensional (3D), scalable, and fully defined thermoresponsive biomaterial system. We used CRISPR/Cas9 to create a NKX2.2-EGFP human embryonic stem cell reporter line that enabled fine-tuning of early OPC specification and identification of conditions that markedly increased the number of OLIG2+ and NKX2.2+ cells generated from hPSCs. Transplantation of 50-day-old OPCs into the brains of NOD/SCID mice revealed that progenitors generated in 3D without cell selection or purification subsequently engrafted, migrated, and matured into myelinating oligodendrocytes in vivo. These results demonstrate the potential of harnessing lineage reporter lines to develop 3D platforms for rapid and large-scale production of OPCs.
AB - Oligodendrocyte precursor cells (OPCs) offer considerable potential for the treatment of demyelinating diseases and injuries of the CNS. However, generating large quantities of high-quality OPCs remains a substantial challenge that impedes their therapeutic application. Here, we show that OPCs can be generated from human pluripotent stem cells (hPSCs) in a three-dimensional (3D), scalable, and fully defined thermoresponsive biomaterial system. We used CRISPR/Cas9 to create a NKX2.2-EGFP human embryonic stem cell reporter line that enabled fine-tuning of early OPC specification and identification of conditions that markedly increased the number of OLIG2+ and NKX2.2+ cells generated from hPSCs. Transplantation of 50-day-old OPCs into the brains of NOD/SCID mice revealed that progenitors generated in 3D without cell selection or purification subsequently engrafted, migrated, and matured into myelinating oligodendrocytes in vivo. These results demonstrate the potential of harnessing lineage reporter lines to develop 3D platforms for rapid and large-scale production of OPCs.
KW - CRISPR/Cas9 knockin
KW - NKX2.2-EGFP reporter cell line
KW - human pluripotent stem cells
KW - oligodendrocyte precursor cells
KW - scalable and defined oligodendrocyte differentiation
KW - three-dimensional culture systems
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U2 - 10.1016/j.stemcr.2017.04.027
DO - 10.1016/j.stemcr.2017.04.027
M3 - Article
C2 - 28552605
AN - SCOPUS:85019626381
SN - 2213-6711
VL - 8
SP - 1770
EP - 1783
JO - Stem Cell Reports
JF - Stem Cell Reports
IS - 6
ER -