TY - JOUR
T1 - Murine cerebral organoids develop network of functional neurons and hippocampal brain region identity
AU - Ciarpella, Francesca
AU - Zamfir, Raluca Georgiana
AU - Campanelli, Alessandra
AU - Ren, Elisa
AU - Pedrotti, Giulia
AU - Bottani, Emanuela
AU - Borioli, Andrea
AU - Caron, Davide
AU - Di Chio, Marzia
AU - Dolci, Sissi
AU - Ahtiainen, Annika
AU - Malpeli, Giorgio
AU - Malerba, Giovanni
AU - Bardoni, Rita
AU - Fumagalli, Guido
AU - Hyttinen, Jari
AU - Bifari, Francesco
AU - Palazzolo, Gemma
AU - Panuccio, Gabriella
AU - Curia, Giulia
AU - Decimo, Ilaria
N1 - Funding Information:
We acknowledge Dr. Edward Callaway, Systems Neurobiology Laboratories, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA, for providing the modified rabies virus particle, Margrate Anyanwu (University of Verona) for help with immunofluorescence staining, Dr. Monica Castellucci, CPT University of Verona, for technical help with RNAseq, Dr. Giacomo Pruzzo, IIT Genoa, for MEA mold design and realization of the custom mini-incubator. ?Centro Interdipartimentale di Servizi per la Ricerca che utilizza Animali da Laboratorio?? C.I.R.S.A.L. and ?Centro PiattaformeTecnologiche??CPT (University of Verona) are acknowledged for services and support. European Union project FETPROACT-2018-2020 HERMES [grant number 824164], is acknowledged for the support on research provided to I.D, G.C. G.P. and G.P. J.H.; Fondazione Telethon?Italy (Grant Number GSP20004_PAsMCT8006) to I.D. Italian patient association la Colonna and GALM and University of Verona (Grant Number DDSP-FUR-6616) to I.D. and G.F.; University of Milan (Grant Number BIOMETRA15-6-3003005-1 and PSR2018_RIVA_BIFARI) to F.B; and Fondazione Telethon?Italy (Grant Number GGP19250) to F.B. F.C. R.G.Z. and A.C. performed experiments, analyzed and interpreted data; E.R. performed electrophysiological experiments, analyzed and interpreted data; G.P. E.B. performed metabolic experiments, analyzed and interpreted data; A.B. M.D.C. S.D. performed immunofluorescence experiments; D.C. performed MEA recording and interpreted data; A.A. J.H. performed OPT microscopy and organoid volume analysis; G. M. G.M. analyzed and interpreted gene expression data; R.B. performed electrophysiological experiments; G.F. F.B. G.P. G.P. G.C. analyzed and interpreted data. F.C. G.C. I.D wrote the paper. All authors discussed results and commented on the manuscript. F.C. R.G.Z. I.D conceptualized the study and I.D. supervised the project and had the scientific direction. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Funding Information:
We acknowledge Dr. Edward Callaway, Systems Neurobiology Laboratories, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA, for providing the modified rabies virus particle, Margrate Anyanwu (University of Verona) for help with immunofluorescence staining, Dr. Monica Castellucci, CPT University of Verona, for technical help with RNAseq, Dr. Giacomo Pruzzo, IIT Genoa, for MEA mold design and realization of the custom mini-incubator. “Centro Interdipartimentale di Servizi per la Ricerca che utilizza Animali da Laboratorio”– C.I.R.S.A.L. and “Centro PiattaformeTecnologiche”–CPT (University of Verona) are acknowledged for services and support. European Union project FETPROACT-2018-2020 HERMES [grant number 824164 ], is acknowledged for the support on research provided to I.D, G.C., G.P. and G.P., J.H.; Fondazione Telethon–Italy (Grant Number GSP20004_PAsMCT8006 ) to I.D. Italian patient association la Colonna and GALM and University of Verona (Grant Number DDSP-FUR-6616 ) to I.D. and G.F.; University of Milan (Grant Number BIOMETRA15-6-3003005-1 and PSR2018_RIVA_BIFARI ) to F.B; and Fondazione Telethon–Italy (Grant Number GGP19250 ) to F.B.
Publisher Copyright:
© 2021 The Author(s)
PY - 2021/12/17
Y1 - 2021/12/17
N2 - Brain organoids are in vitro three-dimensional (3D) self-organized neural structures, which can enable disease modeling and drug screening. However, their use for standardized large-scale drug screening studies is limited by their high batch-to-batch variability, long differentiation time (10–20 weeks), and high production costs. This is particularly relevant when brain organoids are obtained from human induced pluripotent stem cells (iPSCs). Here, we developed, for the first time, a highly standardized, reproducible, and fast (5 weeks) murine brain organoid model starting from embryonic neural stem cells. We obtained brain organoids, which progressively differentiated and self-organized into 3D networks of functional neurons with dorsal forebrain phenotype. Furthermore, by adding the morphogen WNT3a, we generated brain organoids with specific hippocampal region identity. Overall, our results showed the establishment of a fast, robust and reproducible murine 3D in vitro brain model that may represent a useful tool for high-throughput drug screening and disease modeling.
AB - Brain organoids are in vitro three-dimensional (3D) self-organized neural structures, which can enable disease modeling and drug screening. However, their use for standardized large-scale drug screening studies is limited by their high batch-to-batch variability, long differentiation time (10–20 weeks), and high production costs. This is particularly relevant when brain organoids are obtained from human induced pluripotent stem cells (iPSCs). Here, we developed, for the first time, a highly standardized, reproducible, and fast (5 weeks) murine brain organoid model starting from embryonic neural stem cells. We obtained brain organoids, which progressively differentiated and self-organized into 3D networks of functional neurons with dorsal forebrain phenotype. Furthermore, by adding the morphogen WNT3a, we generated brain organoids with specific hippocampal region identity. Overall, our results showed the establishment of a fast, robust and reproducible murine 3D in vitro brain model that may represent a useful tool for high-throughput drug screening and disease modeling.
KW - Biological sciences
KW - Cell biology
KW - Developmental biology
KW - Neuroscience
U2 - 10.1016/j.isci.2021.103438
DO - 10.1016/j.isci.2021.103438
M3 - Article
AN - SCOPUS:85120955204
SN - 2589-0042
VL - 24
JO - Iscience
JF - Iscience
IS - 12
M1 - 103438
ER -