TY - JOUR
T1 - Directed differentiation of notochord-like and nucleus pulposus-like cells using human pluripotent stem cells
AU - Zhang, Y.L.
AU - Zhang, Z.
AU - Chen, P.K.
AU - Ma, C.Y.
AU - Li, C.
AU - Au, T.Y.K.
AU - Tam, V.
AU - Peng, Y.
AU - Wu, R.
AU - Cheung, K.M.C.
AU - Sham, P.C.
AU - Tse, H.F.
AU - Chan, D.
AU - Leung, V.Y.
AU - Cheah, K.S.E.
AU - Lian, Q.Z.
PY - 2020
Y1 - 2020
N2 - Intervertebral disc degeneration might be amenable to stem cell therapy, but the required cells are scarce. Here, we report the development of a protocol for directed in vitro differentiation of human pluripotent stem cells (hPSCs) into notochord-like and nucleus pulposus (NP)-like cells of the disc. The first step combines enhancement of ACTIVIN/NODAL and WNT and inhibition of BMP pathways. By day 5 of differentiation, hPSC-derived cells express notochordal cell characteristic genes. After activating the TGF-beta pathway for an additional 15 days, qPCR, immunostaining, and transcriptome data show that a wide array of NP markers are expressed. Transcriptomically, the in vitro-derived cells become more like in vivo adolescent human NP cells, driven by a set of influential genes enriched with motifs bound by BRACHYURY and FOXA2, consistent with an NP cell-like identity. Transplantation of these NP-like cells attenuates fibrotic changes in a rat disc injury model of disc degeneration.
AB - Intervertebral disc degeneration might be amenable to stem cell therapy, but the required cells are scarce. Here, we report the development of a protocol for directed in vitro differentiation of human pluripotent stem cells (hPSCs) into notochord-like and nucleus pulposus (NP)-like cells of the disc. The first step combines enhancement of ACTIVIN/NODAL and WNT and inhibition of BMP pathways. By day 5 of differentiation, hPSC-derived cells express notochordal cell characteristic genes. After activating the TGF-beta pathway for an additional 15 days, qPCR, immunostaining, and transcriptome data show that a wide array of NP markers are expressed. Transcriptomically, the in vitro-derived cells become more like in vivo adolescent human NP cells, driven by a set of influential genes enriched with motifs bound by BRACHYURY and FOXA2, consistent with an NP cell-like identity. Transplantation of these NP-like cells attenuates fibrotic changes in a rat disc injury model of disc degeneration.
UR - https://hdl.handle.net/1959.7/uws:66771
U2 - 10.1016/j.celrep.2020.01.100
DO - 10.1016/j.celrep.2020.01.100
M3 - Article
SN - 2211-1247
VL - 30
SP - 2791
EP - 2806
JO - Cell Reports
JF - Cell Reports
IS - 8
ER -
