Directed differentiation of notochord-like and nucleus pulposus-like cells using human pluripotent stem cells

Y.L. Zhang; Z. Zhang; P.K. Chen; C.Y. Ma; C. Li; T.Y.K. Au; V. Tam; Y. Peng; R. Wu; K.M.C. Cheung; P.C. Sham; H.F. Tse; D. Chan; V.Y. Leung; K.S.E. Cheah; Q.Z. Lian

doi:10.1016/j.celrep.2020.01.100

Directed differentiation of notochord-like and nucleus pulposus-like cells using human pluripotent stem cells

Y.L. Zhang
, Z. Zhang
, P.K. Chen
, C.Y. Ma
, C. Li
, T.Y.K. Au
, V. Tam
, Y. Peng
, R. Wu
, K.M.C. Cheung
, P.C. Sham
, H.F. Tse
, D. Chan
, V.Y. Leung
, K.S.E. Cheah
, Q.Z. Lian

Research output: Contribution to journal › Article › peer-review

67 Citations (Scopus)

Abstract

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-β 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. Zhang et al. report notochord-like and nucleus pulposus (NP)-like cells can be derived from human pluripotent stem cells using a NOTO-eGFP reporter system and a compound-defined protocol. These derived NP-like cells share high similarities with adolescent human NP cells and attenuate injury-induced intervertebral disc degeneration after transplantation.

Original language	English
Pages (from-to)	2791-2806.e5
Number of pages	16
Journal	Cell Reports
Volume	30
Issue number	8
DOIs	https://doi.org/10.1016/j.celrep.2020.01.100
Publication status	Published - 25 Feb 2020

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Publisher Copyright:
© 2020 The Author(s)

Open Access - Access Right Statement

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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10.1016/j.celrep.2020.01.100

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Zhang, Y. L., Zhang, Z., Chen, P. K., Ma, C. Y., Li, C., Au, T. Y. K., Tam, V., Peng, Y., Wu, R., Cheung, K. M. C., Sham, P. C., Tse, H. F., Chan, D., Leung, V. Y., Cheah, K. S. E., & Lian, Q. Z. (2020). Directed differentiation of notochord-like and nucleus pulposus-like cells using human pluripotent stem cells. Cell Reports, 30(8), 2791-2806.e5. https://doi.org/10.1016/j.celrep.2020.01.100

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abstract = "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-β 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. Zhang et al. report notochord-like and nucleus pulposus (NP)-like cells can be derived from human pluripotent stem cells using a NOTO-eGFP reporter system and a compound-defined protocol. These derived NP-like cells share high similarities with adolescent human NP cells and attenuate injury-induced intervertebral disc degeneration after transplantation.",

author = "Y.L. Zhang and Z. Zhang and P.K. Chen and C.Y. Ma and C. Li and T.Y.K. Au and V. Tam and Y. Peng and R. Wu and K.M.C. Cheung and P.C. Sham and H.F. Tse and D. Chan and V.Y. Leung and K.S.E. Cheah and Q.Z. Lian",

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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.

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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-β 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. Zhang et al. report notochord-like and nucleus pulposus (NP)-like cells can be derived from human pluripotent stem cells using a NOTO-eGFP reporter system and a compound-defined protocol. These derived NP-like cells share high similarities with adolescent human NP cells and attenuate injury-induced intervertebral disc degeneration after transplantation.

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ER -

Directed differentiation of notochord-like and nucleus pulposus-like cells using human pluripotent stem cells

Abstract

Bibliographical note

Open Access - Access Right Statement

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