Direct Reprogramming and Induction of Human Dermal Fibroblasts to Differentiate into iPS-Derived Nucleus Pulposus-like Cells in 3D Culture

Shoji Seki; Mami Iwasaki; Hiroto Makino; Yasuhito Yahara; Yoshitaka Miyazaki; Katsuhiko Kamei; Hayato Futakawa; Makiko Nogami; Nguyen Tran Canh Tung; Tatsuro Hirokawa; Mamiko Tsuji; Yoshiharu Kawaguchi

doi:10.3390/ijms23074059

Direct Reprogramming and Induction of Human Dermal Fibroblasts to Differentiate into iPS-Derived Nucleus Pulposus-like Cells in 3D Culture

Shoji Seki^*, Mami Iwasaki, Hiroto Makino, Yasuhito Yahara, Yoshitaka Miyazaki, Katsuhiko Kamei, Hayato Futakawa, Makiko Nogami, Nguyen Tran Canh Tung, Tatsuro Hirokawa, Mamiko Tsuji, Yoshiharu Kawaguchi

^*Corresponding author for this work

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

5 Scopus citations

Abstract

Intervertebral disc (IVD) diseases are common spinal disorders that cause neck or back pain in the presence or absence of an underlying neurological disorder. IVD diseases develop on the basis of degeneration, and there are no established treatments for degeneration. IVD diseases may therefore represent a candidate for the application of regenerative medicine, potentially employing normal human dermal fibroblasts (NHDFs) induced to differentiate into nucleus pulposus (NP) cells. Here, we used a three-dimensional culture system to demonstrate that ectopic expression of MYC, KLF4, NOTO, SOX5, SOX6, and SOX9 in NHDFs generated NP-like cells, detected using Safranin-O staining. Quantitative PCR, microarray analysis, and fluorescence-activated cell sorting revealed that the induced NP cells exhibited a fully differentiated phenotype. These findings may significantly contribute to the development of effective strategies for treating IVD diseases.

Original language	English
Article number	4059
Journal	International Journal of Molecular Sciences
Volume	23
Issue number	7
DOIs	https://doi.org/10.3390/ijms23074059
State	Published - 2022/04/01

Keywords

alginate beads
direct reprogramming
human dermal fibroblast
induced pluripotent stem cells
intervertebral disc disease
intervertebral disc regeneration
nucleus pulposus cell

ASJC Scopus subject areas

Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry

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10.3390/ijms23074059

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Seki, S., Iwasaki, M., Makino, H., Yahara, Y., Miyazaki, Y., Kamei, K., Futakawa, H., Nogami, M., Tung, N. T. C., Hirokawa, T., Tsuji, M., & Kawaguchi, Y. (2022). Direct Reprogramming and Induction of Human Dermal Fibroblasts to Differentiate into iPS-Derived Nucleus Pulposus-like Cells in 3D Culture. International Journal of Molecular Sciences, 23(7), Article 4059. https://doi.org/10.3390/ijms23074059

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title = "Direct Reprogramming and Induction of Human Dermal Fibroblasts to Differentiate into iPS-Derived Nucleus Pulposus-like Cells in 3D Culture",

abstract = "Intervertebral disc (IVD) diseases are common spinal disorders that cause neck or back pain in the presence or absence of an underlying neurological disorder. IVD diseases develop on the basis of degeneration, and there are no established treatments for degeneration. IVD diseases may therefore represent a candidate for the application of regenerative medicine, potentially employing normal human dermal fibroblasts (NHDFs) induced to differentiate into nucleus pulposus (NP) cells. Here, we used a three-dimensional culture system to demonstrate that ectopic expression of MYC, KLF4, NOTO, SOX5, SOX6, and SOX9 in NHDFs generated NP-like cells, detected using Safranin-O staining. Quantitative PCR, microarray analysis, and fluorescence-activated cell sorting revealed that the induced NP cells exhibited a fully differentiated phenotype. These findings may significantly contribute to the development of effective strategies for treating IVD diseases.",

keywords = "alginate beads, direct reprogramming, human dermal fibroblast, induced pluripotent stem cells, intervertebral disc disease, intervertebral disc regeneration, nucleus pulposus cell",

author = "Shoji Seki and Mami Iwasaki and Hiroto Makino and Yasuhito Yahara and Yoshitaka Miyazaki and Katsuhiko Kamei and Hayato Futakawa and Makiko Nogami and Tung, {Nguyen Tran Canh} and Tatsuro Hirokawa and Mamiko Tsuji and Yoshiharu Kawaguchi",

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year = "2022",

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Seki, S , Iwasaki, M , Makino, H, Yahara, Y, Miyazaki, Y, Kamei, K, Futakawa, H, Nogami, M, Tung, NTC, Hirokawa, T, Tsuji, M & Kawaguchi, Y 2022, 'Direct Reprogramming and Induction of Human Dermal Fibroblasts to Differentiate into iPS-Derived Nucleus Pulposus-like Cells in 3D Culture', International Journal of Molecular Sciences, vol. 23, no. 7, 4059. https://doi.org/10.3390/ijms23074059

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T1 - Direct Reprogramming and Induction of Human Dermal Fibroblasts to Differentiate into iPS-Derived Nucleus Pulposus-like Cells in 3D Culture

AU - Seki, Shoji

AU - Iwasaki, Mami

AU - Makino, Hiroto

AU - Yahara, Yasuhito

AU - Miyazaki, Yoshitaka

AU - Kamei, Katsuhiko

AU - Futakawa, Hayato

AU - Nogami, Makiko

AU - Tung, Nguyen Tran Canh

AU - Hirokawa, Tatsuro

AU - Tsuji, Mamiko

AU - Kawaguchi, Yoshiharu

PY - 2022/4/1

Y1 - 2022/4/1

N2 - Intervertebral disc (IVD) diseases are common spinal disorders that cause neck or back pain in the presence or absence of an underlying neurological disorder. IVD diseases develop on the basis of degeneration, and there are no established treatments for degeneration. IVD diseases may therefore represent a candidate for the application of regenerative medicine, potentially employing normal human dermal fibroblasts (NHDFs) induced to differentiate into nucleus pulposus (NP) cells. Here, we used a three-dimensional culture system to demonstrate that ectopic expression of MYC, KLF4, NOTO, SOX5, SOX6, and SOX9 in NHDFs generated NP-like cells, detected using Safranin-O staining. Quantitative PCR, microarray analysis, and fluorescence-activated cell sorting revealed that the induced NP cells exhibited a fully differentiated phenotype. These findings may significantly contribute to the development of effective strategies for treating IVD diseases.

AB - Intervertebral disc (IVD) diseases are common spinal disorders that cause neck or back pain in the presence or absence of an underlying neurological disorder. IVD diseases develop on the basis of degeneration, and there are no established treatments for degeneration. IVD diseases may therefore represent a candidate for the application of regenerative medicine, potentially employing normal human dermal fibroblasts (NHDFs) induced to differentiate into nucleus pulposus (NP) cells. Here, we used a three-dimensional culture system to demonstrate that ectopic expression of MYC, KLF4, NOTO, SOX5, SOX6, and SOX9 in NHDFs generated NP-like cells, detected using Safranin-O staining. Quantitative PCR, microarray analysis, and fluorescence-activated cell sorting revealed that the induced NP cells exhibited a fully differentiated phenotype. These findings may significantly contribute to the development of effective strategies for treating IVD diseases.

KW - alginate beads

KW - direct reprogramming

KW - human dermal fibroblast

KW - induced pluripotent stem cells

KW - intervertebral disc disease

KW - intervertebral disc regeneration

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ER -

Direct Reprogramming and Induction of Human Dermal Fibroblasts to Differentiate into iPS-Derived Nucleus Pulposus-like Cells in 3D Culture

Abstract

Keywords

ASJC Scopus subject areas

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