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.
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea; Department of Biomaterials Science and Engineering, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Kisuk
authors.initials: K
authors.lastname: Yang
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Hyun-Ji
authors.initials: HJ
authors.lastname: Park
authors.affiliation: School of Mechanical Engineering, Korea University, Seoul 136-713, Republic of Korea.
authors.forename: Sewoon
authors.initials: S
authors.lastname: Han
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Joan
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Eunkyung
authors.initials: E
authors.lastname: Ko
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Jin
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Jong Seung
authors.initials: JS
authors.lastname: Lee
authors.affiliation: Department and Research Institute of Rehabilitation Medicine, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.
authors.forename: Ji Hea
authors.initials: JH
authors.lastname: Yu
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Ki Yeong
authors.initials: KY
authors.lastname: Song
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Eunji
authors.initials: E
authors.lastname: Cheong
authors.affiliation: Department and Research Institute of Rehabilitation Medicine, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.
authors.forename: Sung-Rae
authors.initials: SR
authors.lastname: Cho
authors.affiliation: School of Mechanical Engineering, Korea University, Seoul 136-713, Republic of Korea.
authors.forename: Seok
authors.initials: S
authors.lastname: Chung
authors.affiliation: Department of Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea; Department of Neurosurgery, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea. Electronic address: seungwoocho@yonsei.ac.kr.
authors.forename: Seung-Woo
authors.initials: SW
authors.lastname: Cho
mesh_terms: D000818:Animals; D001930:Brain Injuries / Q000628:therapy; D018929:Cell Culture Techniques / Q000295:instrumentation*; D004867:Equipment Design; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D006801:Humans; D056656:Lab-On-A-Chip Devices; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D055495:Neurogenesis; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D019899:Paracrine Communication; D015854:Up-Regulation
chemical_list: D051100:Glial Cell Line-Derived Neurotrophic Factor
keywords: Glial cell-derived neurotrophic factor; Human mesenchymal stem cell; Human neural stem cell; Microfluidic array; Neuronal differentiation; Paracrine signal