TY - JOUR
T1 - Simultaneous induction of vasculature and neuronal network formation on a chip reveals a dynamic interrelationship between cell types
AU - Isosaari, Lotta
AU - Vuorenpää, Hanna
AU - Yrjänäinen, Alma
AU - Kapucu, Fikret Emre
AU - Kelloniemi, Minna
AU - Pakarinen, Toni Karri
AU - Miettinen, Susanna
AU - Narkilahti, Susanna
N1 - Funding Information:
We thank Hanna Mäkelä, Eija Hannuksela, Anna-Maija Honkala and Sari Kalliokoski for their technical assistance. The authors acknowledge the Tampere Imaging Facility (TIF), the Tampere Facility of iPS Cells, and the Tampere CellTech Laboratories for their service. The authors also thank Biocenter Finland for the support of Imaging and iPS cells facilities.
Funding Information:
Open access funding provided by Tampere University including Tampere University Hospital, Tampere University of Applied Sciences (TUNI). This research was funded by the Academy of Finland, the Center of Excellence in Body-on-Chip Research (336665 to SN, 336666 to SM), Academy of Finland postdoctoral project (332693 to FEK), State Research Financing of the Tampere University Hospital Specific Catchment Area (SM), and Doctoral Programme Funding at the Faculty of Medicine and Health Technology at Tampere University (AY).
Publisher Copyright:
© 2023, The Author(s).
PY - 2023
Y1 - 2023
N2 - Background: Neuronal networks receive and deliver information to regulate bodily functions while the vascular network provides oxygen, nutrients, and signaling molecules to tissues. Neurovascular interactions are vital for both tissue development and maintaining homeostasis in adulthood; these two network systems align and reciprocally communicate with one another. Although communication between network systems has been acknowledged, the lack of relevant in vitro models has hindered research at the mechanistic level. For example, the current used in vitro neurovascular models are typically established to be short-term (≤ 7 days) culture models, and they miss the supporting vascular mural cells. Methods: In this study, we utilized human induced pluripotent stem cell (hiPSC) -derived neurons, fluorescence tagged human umbilical vein endothelial cells (HUVECs), and either human bone marrow or adipose stem/stromal cells (BMSCs or ASCs) as the mural cell types to create a novel 3D neurovascular network-on-a-chip model. Collagen 1–fibrin matrix was used to establish long-term (≥ 14 days) 3D cell culture in a perfusable microphysiological environment. Results: Aprotinin-supplemented endothelial cell growth medium-2 (EGM-2) supported the simultaneous formation of neuronal networks, vascular structures, mural cell differentiation, and the stability of the 3D matrix. The formed neuronal and vascular networks were morphologically and functionally characterized. Neuronal networks supported vasculature formation based on direct cell contacts and by dramatically increasing the secretion of angiogenesis-related factors in multicultures in contrast to cocultures without neurons. Both utilized mural cell types supported the formation of neurovascular networks; however, the BMSCs seemed to boost neurovascular networks to greater extent. Conclusions: Overall, our study provides a novel human neurovascular network model that is applicable for creating in vivo-like tissue models with intrinsic neurovascular interactions. The 3D neurovascular network model on chip forms an initial platform for the development of vascularized and innervated organ-on-chip and further body-on-chip concepts and offers the possibility for mechanistic studies on neurovascular communication both under healthy and in disease conditions. [MediaObject not available: see fulltext.]
AB - Background: Neuronal networks receive and deliver information to regulate bodily functions while the vascular network provides oxygen, nutrients, and signaling molecules to tissues. Neurovascular interactions are vital for both tissue development and maintaining homeostasis in adulthood; these two network systems align and reciprocally communicate with one another. Although communication between network systems has been acknowledged, the lack of relevant in vitro models has hindered research at the mechanistic level. For example, the current used in vitro neurovascular models are typically established to be short-term (≤ 7 days) culture models, and they miss the supporting vascular mural cells. Methods: In this study, we utilized human induced pluripotent stem cell (hiPSC) -derived neurons, fluorescence tagged human umbilical vein endothelial cells (HUVECs), and either human bone marrow or adipose stem/stromal cells (BMSCs or ASCs) as the mural cell types to create a novel 3D neurovascular network-on-a-chip model. Collagen 1–fibrin matrix was used to establish long-term (≥ 14 days) 3D cell culture in a perfusable microphysiological environment. Results: Aprotinin-supplemented endothelial cell growth medium-2 (EGM-2) supported the simultaneous formation of neuronal networks, vascular structures, mural cell differentiation, and the stability of the 3D matrix. The formed neuronal and vascular networks were morphologically and functionally characterized. Neuronal networks supported vasculature formation based on direct cell contacts and by dramatically increasing the secretion of angiogenesis-related factors in multicultures in contrast to cocultures without neurons. Both utilized mural cell types supported the formation of neurovascular networks; however, the BMSCs seemed to boost neurovascular networks to greater extent. Conclusions: Overall, our study provides a novel human neurovascular network model that is applicable for creating in vivo-like tissue models with intrinsic neurovascular interactions. The 3D neurovascular network model on chip forms an initial platform for the development of vascularized and innervated organ-on-chip and further body-on-chip concepts and offers the possibility for mechanistic studies on neurovascular communication both under healthy and in disease conditions. [MediaObject not available: see fulltext.]
KW - 3D cell culture
KW - Angiogenesis
KW - Cellular communication
KW - Human cells
KW - Microfluidic
KW - Neurovascular interactions
U2 - 10.1186/s12964-023-01159-4
DO - 10.1186/s12964-023-01159-4
M3 - Article
C2 - 37316873
AN - SCOPUS:85161899554
SN - 1478-811X
VL - 21
JO - CELL COMMUNICATION AND SIGNALING
JF - CELL COMMUNICATION AND SIGNALING
M1 - 132
ER -