3D-printed microplate inserts for long term high-resolution imaging of live brain organoids

BMC Biomedical Engineering

Sally L. Perrin¹  Sakthi Lenin¹  Guillermo A. Gomez¹  Kristyna Sedivakova¹  Gökhan Cildir¹  Elise Ponthier¹  Mariana Oksdath Mansilla¹  Erica C. F. Yeo¹  Kaitlin G. Scheer¹  Melinda N. Tea¹  Camilo Salazar-Hernandez¹  John Toubia²  Vinay Tergaonkar³  Stuart M. Pitson⁴  Lisa M. Ebert⁵  Michael P. Brown⁵  Santosh Poonnoose⁶  Rebecca J. Ormsby⁷ 

[1] Centre for Cancer Biology, SA Pathology and University of South Australia, 5000, Adelaide, SA, Australia;Centre for Cancer Biology, SA Pathology and University of South Australia, 5000, Adelaide, SA, Australia;ACRF Cancer Genomics Facility, Centre for Cancer Biology, SA Pathology and University of South Australia, Frome Road, 5000, Adelaide, SA, Australia;Centre for Cancer Biology, SA Pathology and University of South Australia, 5000, Adelaide, SA, Australia;Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A-STAR), Singapore, Singapore;Department of Pathology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore;Centre for Cancer Biology, SA Pathology and University of South Australia, 5000, Adelaide, SA, Australia;School of Medicine, University of Adelaide, 5000, Adelaide, SA, Australia;Centre for Cancer Biology, SA Pathology and University of South Australia, 5000, Adelaide, SA, Australia;School of Medicine, University of Adelaide, 5000, Adelaide, SA, Australia;Cancer Clinical Trials Unit, Royal Adelaide Hospital, 5000, Adelaide, SA, Australia;Department of Neurosurgery, Flinders Medical Centre, 5042, Adelaide, SA, Australia;Flinders Health & Medical Research Institute, College of Medicine & Public Health, Flinders University, 5042, Adelaide, SA, Australia;Flinders Health & Medical Research Institute, College of Medicine & Public Health, Flinders University, 5042, Adelaide, SA, Australia;

关键词: Brain organoids;    Live-imaging;    Fluorescence microscopy;    Glioblastoma;   

DOI  :  10.1186/s42490-021-00049-5

来源: Springer

【 摘 要 】

BackgroundOrganoids are a reliable model used in the study of human brain development and under pathological conditions. However, current methods for brain organoid culture generate tissues that range from 0.5 to 2 mm of size, which need to be constantly agitated to allow proper oxygenation. The culture conditions are, therefore, not suitable for whole-brain organoid live imaging, required to study developmental processes and disease progression within physiologically relevant time frames (i.e. days, weeks, months).ResultsHere we designed 3D-printed microplate inserts adaptable to standard 24 multi-well plates, which allow the growth of multiple organoids in pre-defined and fixed XYZ coordinates. This innovation facilitates high-resolution imaging of whole-cerebral organoids, allowing precise assessment of organoid growth and morphology, as well as cell tracking within the organoids, over long periods. We applied this technology to track neocortex development through neuronal progenitors in brain organoids, as well as the movement of patient-derived glioblastoma stem cells within healthy brain organoids.ConclusionsThis new bioengineering platform constitutes a significant advance that permits long term detailed analysis of whole-brain organoids using multimodal inverted fluorescence microscopy.

【 授权许可】

CC BY