Protocols for 3D Models and Techniques
Generation of Forebrain Spheroids
This protocol describes how to generate forebrain spheroids from iPSCs. Forebrain spheroids are formed in stationary cultures, they contain cortical neurons and astrocytes. These spheroids can be maintained in cultures for several months allowing the study of neuronal progenitors and mature cells.
Cryosectioning of fixed and frozen tissue
This protocol will provide a standardized procedure on methods developed for cryosectioning of fixed or fresh frozen tissue in Optimal Cutting Temperature (OCT) using a motorized cryostat. This SOP also outlines minimum assessment criteria that should be in place to evaluate the quality and integrity of frozen tissue sections and troubleshooting steps when issues with sectioning arise.
Human Midbrain Organoid (hMO) Generation with EB Disks
At the EDDU, we developed a method with microfabricated disk (EB disk-948 from eNUVIO) to scale up the generation of hMOs (human midbrain organoids). This opens up the possibility to generate larger numbers of hMOs, in a manner that minimizes the amount of labor required, while decreasing variability and maintaining the viability of these hMOs over time.
This video will describe the methodology to generate hMOs from a single EB-Disk containing 948 microwells. Visit this link to access the published version of this protocol by Mohamed et al.