Recapitulation of in vivo-like paracrine signals of human mesenchymal stem cells for functional neuronal differentiation of human neural stem cells in a 3D microfluidic system

Kisuk Yang; Hyun Ji Park; Sewoon Han; Joan Lee; Eunkyung Ko; Jin Kim; Jong Seung Lee; Ji Hea Yu; Ki Yeong Song; Eunji Cheong; Sung Rae Cho; Seok Chung; Seung Woo Cho

doi:10.1016/j.biomaterials.2015.06.011

Recapitulation of in vivo-like paracrine signals of human mesenchymal stem cells for functional neuronal differentiation of human neural stem cells in a 3D microfluidic system

Kisuk Yang, Hyun Ji Park, Sewoon Han, Joan Lee, Eunkyung Ko, Jin Kim, Jong Seung Lee, Ji Hea Yu, Ki Yeong Song, Eunji Cheong, Sung Rae Cho, Seok Chung, Seung Woo Cho

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Abstract

Paracrine signals produced from stem cells influence tissue regeneration by inducing the differentiation of endogenous stem or progenitor cells. However, many recent studies that have investigated paracrine signaling of stem cells have relied on either two-dimensional transwell systems or conditioned medium culture, neither of which provide optimal culture microenvironments for elucidating the effects of paracrine signals in vivo. In this study, we recapitulated in vivo-like paracrine signaling of human mesenchymal stem cells (hMSCs) to enhance functional neuronal differentiation of human neural stem cells (hNSCs) in three-dimensional (3D) extracellular matrices (ECMs) within a microfluidic array platform. In order to amplify paracrine signaling, hMSCs were genetically engineered using cationic polymer nanoparticles to overexpress glial cell-derived neurotrophic factor (GDNF). hNSCs were cultured in 3D ECM hydrogel used to fill central channels of the microfluidic device, while GDNF-overexpressing hMSCs (GDNF-hMSCs) were cultured in channels located on both sides of the central channel. This setup allowed for mimicking of paracrine signaling between genetically engineered hMSCs and endogenous hNSCs in the brain. Co-culture of hNSCs with GDNF-hMSCs in the 3D microfluidic system yielded reduced glial differentiation of hNSCs while significantly enhancing differentiation into neuronal cells including dopaminergic neurons. Neuronal cells produced from hNSCs differentiating in the presence of GDNF-hMSCs exhibited functional neuron-like electrophysiological features. The enhanced paracrine ability of GDNF-hMSCs was finally confirmed using an animal model of hypoxic-ischemic brain injury. This study demonstrates the presented 3D microfluidic array device can provide an efficient co-culture platform and provide an environment for paracrine signals from transplanted stem cells to control endogenous neuronal behaviors in vivo.

Original language	English
Pages (from-to)	177-188
Number of pages	12
Journal	Biomaterials
Volume	63
DOIs	https://doi.org/10.1016/j.biomaterials.2015.06.011
Publication status	Published - 2015 Sept 1

Bibliographical note

Funding Information:
This study was supported by grants ( NRF-2010-0020409 , NRF-2010-0020408 , and NRF-2013R1A1A2A10061422 ) from the National Research Foundation of Korea (NRF) , Republic of Korea. This work was also supported in part by a grant ( HI14C1588 ) from the Korea Health Technology R&D Project of the Ministry of Health & Welfare and the Yonsei University Future-Leading Research Initiative of 2014. Kisuk Yang was supported by an NRF grant (NRF-2013-Fostering Core Leaders of the Future Basic Science Program). Prof. Seok Chung was supported by Nano Material Technology Development Program from NRF ( 2014M3A7B4052193 ), Republic of Korea.

Publisher Copyright:
© 2015 Elsevier Ltd.

All Science Journal Classification (ASJC) codes

Biophysics
Bioengineering
Ceramics and Composites
Biomaterials
Mechanics of Materials

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10.1016/j.biomaterials.2015.06.011

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Yang, K., Park, H. J., Han, S., Lee, J., Ko, E., Kim, J., Lee, J. S., Yu, J. H., Song, K. Y., Cheong, E., Cho, S. R., Chung, S., & Cho, S. W. (2015). Recapitulation of in vivo-like paracrine signals of human mesenchymal stem cells for functional neuronal differentiation of human neural stem cells in a 3D microfluidic system. Biomaterials, 63, 177-188. https://doi.org/10.1016/j.biomaterials.2015.06.011

@article{070165494a684378acde35ba0fb2ea9a,

title = "Recapitulation of in vivo-like paracrine signals of human mesenchymal stem cells for functional neuronal differentiation of human neural stem cells in a 3D microfluidic system",

abstract = "Paracrine signals produced from stem cells influence tissue regeneration by inducing the differentiation of endogenous stem or progenitor cells. However, many recent studies that have investigated paracrine signaling of stem cells have relied on either two-dimensional transwell systems or conditioned medium culture, neither of which provide optimal culture microenvironments for elucidating the effects of paracrine signals in vivo. In this study, we recapitulated in vivo-like paracrine signaling of human mesenchymal stem cells (hMSCs) to enhance functional neuronal differentiation of human neural stem cells (hNSCs) in three-dimensional (3D) extracellular matrices (ECMs) within a microfluidic array platform. In order to amplify paracrine signaling, hMSCs were genetically engineered using cationic polymer nanoparticles to overexpress glial cell-derived neurotrophic factor (GDNF). hNSCs were cultured in 3D ECM hydrogel used to fill central channels of the microfluidic device, while GDNF-overexpressing hMSCs (GDNF-hMSCs) were cultured in channels located on both sides of the central channel. This setup allowed for mimicking of paracrine signaling between genetically engineered hMSCs and endogenous hNSCs in the brain. Co-culture of hNSCs with GDNF-hMSCs in the 3D microfluidic system yielded reduced glial differentiation of hNSCs while significantly enhancing differentiation into neuronal cells including dopaminergic neurons. Neuronal cells produced from hNSCs differentiating in the presence of GDNF-hMSCs exhibited functional neuron-like electrophysiological features. The enhanced paracrine ability of GDNF-hMSCs was finally confirmed using an animal model of hypoxic-ischemic brain injury. This study demonstrates the presented 3D microfluidic array device can provide an efficient co-culture platform and provide an environment for paracrine signals from transplanted stem cells to control endogenous neuronal behaviors in vivo.",

author = "Kisuk Yang and Park, {Hyun Ji} and Sewoon Han and Joan Lee and Eunkyung Ko and Jin Kim and Lee, {Jong Seung} and Yu, {Ji Hea} and Song, {Ki Yeong} and Eunji Cheong and Cho, {Sung Rae} and Seok Chung and Cho, {Seung Woo}",

note = "Funding Information: This study was supported by grants ( NRF-2010-0020409 , NRF-2010-0020408 , and NRF-2013R1A1A2A10061422 ) from the National Research Foundation of Korea (NRF) , Republic of Korea. This work was also supported in part by a grant ( HI14C1588 ) from the Korea Health Technology R&D Project of the Ministry of Health & Welfare and the Yonsei University Future-Leading Research Initiative of 2014. Kisuk Yang was supported by an NRF grant (NRF-2013-Fostering Core Leaders of the Future Basic Science Program). Prof. Seok Chung was supported by Nano Material Technology Development Program from NRF ( 2014M3A7B4052193 ), Republic of Korea. Publisher Copyright: {\textcopyright} 2015 Elsevier Ltd.",

year = "2015",

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doi = "10.1016/j.biomaterials.2015.06.011",

language = "English",

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Yang, K, Park, HJ, Han, S, Lee, J, Ko, E, Kim, J, Lee, JS, Yu, JH, Song, KY, Cheong, E , Cho, SR, Chung, S & Cho, SW 2015, 'Recapitulation of in vivo-like paracrine signals of human mesenchymal stem cells for functional neuronal differentiation of human neural stem cells in a 3D microfluidic system', Biomaterials, vol. 63, pp. 177-188. https://doi.org/10.1016/j.biomaterials.2015.06.011

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AU - Yang, Kisuk

AU - Park, Hyun Ji

AU - Han, Sewoon

AU - Lee, Joan

AU - Ko, Eunkyung

AU - Kim, Jin

AU - Lee, Jong Seung

AU - Yu, Ji Hea

AU - Song, Ki Yeong

AU - Cheong, Eunji

AU - Cho, Sung Rae

AU - Chung, Seok

AU - Cho, Seung Woo

N1 - Funding Information: This study was supported by grants ( NRF-2010-0020409 , NRF-2010-0020408 , and NRF-2013R1A1A2A10061422 ) from the National Research Foundation of Korea (NRF) , Republic of Korea. This work was also supported in part by a grant ( HI14C1588 ) from the Korea Health Technology R&D Project of the Ministry of Health & Welfare and the Yonsei University Future-Leading Research Initiative of 2014. Kisuk Yang was supported by an NRF grant (NRF-2013-Fostering Core Leaders of the Future Basic Science Program). Prof. Seok Chung was supported by Nano Material Technology Development Program from NRF ( 2014M3A7B4052193 ), Republic of Korea. Publisher Copyright: © 2015 Elsevier Ltd.

PY - 2015/9/1

Y1 - 2015/9/1

N2 - Paracrine signals produced from stem cells influence tissue regeneration by inducing the differentiation of endogenous stem or progenitor cells. However, many recent studies that have investigated paracrine signaling of stem cells have relied on either two-dimensional transwell systems or conditioned medium culture, neither of which provide optimal culture microenvironments for elucidating the effects of paracrine signals in vivo. In this study, we recapitulated in vivo-like paracrine signaling of human mesenchymal stem cells (hMSCs) to enhance functional neuronal differentiation of human neural stem cells (hNSCs) in three-dimensional (3D) extracellular matrices (ECMs) within a microfluidic array platform. In order to amplify paracrine signaling, hMSCs were genetically engineered using cationic polymer nanoparticles to overexpress glial cell-derived neurotrophic factor (GDNF). hNSCs were cultured in 3D ECM hydrogel used to fill central channels of the microfluidic device, while GDNF-overexpressing hMSCs (GDNF-hMSCs) were cultured in channels located on both sides of the central channel. This setup allowed for mimicking of paracrine signaling between genetically engineered hMSCs and endogenous hNSCs in the brain. Co-culture of hNSCs with GDNF-hMSCs in the 3D microfluidic system yielded reduced glial differentiation of hNSCs while significantly enhancing differentiation into neuronal cells including dopaminergic neurons. Neuronal cells produced from hNSCs differentiating in the presence of GDNF-hMSCs exhibited functional neuron-like electrophysiological features. The enhanced paracrine ability of GDNF-hMSCs was finally confirmed using an animal model of hypoxic-ischemic brain injury. This study demonstrates the presented 3D microfluidic array device can provide an efficient co-culture platform and provide an environment for paracrine signals from transplanted stem cells to control endogenous neuronal behaviors in vivo.

AB - Paracrine signals produced from stem cells influence tissue regeneration by inducing the differentiation of endogenous stem or progenitor cells. However, many recent studies that have investigated paracrine signaling of stem cells have relied on either two-dimensional transwell systems or conditioned medium culture, neither of which provide optimal culture microenvironments for elucidating the effects of paracrine signals in vivo. In this study, we recapitulated in vivo-like paracrine signaling of human mesenchymal stem cells (hMSCs) to enhance functional neuronal differentiation of human neural stem cells (hNSCs) in three-dimensional (3D) extracellular matrices (ECMs) within a microfluidic array platform. In order to amplify paracrine signaling, hMSCs were genetically engineered using cationic polymer nanoparticles to overexpress glial cell-derived neurotrophic factor (GDNF). hNSCs were cultured in 3D ECM hydrogel used to fill central channels of the microfluidic device, while GDNF-overexpressing hMSCs (GDNF-hMSCs) were cultured in channels located on both sides of the central channel. This setup allowed for mimicking of paracrine signaling between genetically engineered hMSCs and endogenous hNSCs in the brain. Co-culture of hNSCs with GDNF-hMSCs in the 3D microfluidic system yielded reduced glial differentiation of hNSCs while significantly enhancing differentiation into neuronal cells including dopaminergic neurons. Neuronal cells produced from hNSCs differentiating in the presence of GDNF-hMSCs exhibited functional neuron-like electrophysiological features. The enhanced paracrine ability of GDNF-hMSCs was finally confirmed using an animal model of hypoxic-ischemic brain injury. This study demonstrates the presented 3D microfluidic array device can provide an efficient co-culture platform and provide an environment for paracrine signals from transplanted stem cells to control endogenous neuronal behaviors in vivo.

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Recapitulation of in vivo-like paracrine signals of human mesenchymal stem cells for functional neuronal differentiation of human neural stem cells in a 3D microfluidic system

Abstract

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