Protocol for controlled cortical impact in human cerebral organoids to model traumatic brain injury

Abstract

Summary Modeling traumatic brain injury (TBI) has been a challenge. Rodent and cellular models have provided relevant contributions despite their limitations. Here, we present a protocol for a TBI model based on the controlled cortical impact (CCI) performed on human cerebral organoids (COs), self-assembled 3D cultures that recapitulate features of the human brain. Here, we generate COs from iPSCs obtained from reprogrammed fibroblasts. For complete details on the use and execution of this protocol, please refer to Ramirez et al. (2021) .

Opening the black box of traumatic brain injury: a holistic approach combining human 3D neural tissue and an in vitro traumatic brain injury induction device

“…This is a significant weakness as traumatic brain injuries typically involve structural damage to the brain and affect different types of cells and networks of cellular interaction. To overcome this limitation, bioengineered tissues grown in 3D culture systems, mimicking native brain anatomy and physiological responses, are emerging as powerful in vitro models to investigate the pathophysiology of TBI (Ramirez et al, 2021b ; Silvosa et al, 2022 ). This led to the development of a new in vitro model of TBI combining human cells organized in a 3D neural tissue subjecting to fluid percussion injury.…”

“…Recent technological advancements have allowed the creation of 3D brainlike structures called cerebral organoids (Chiaradia and Lancaster, 2020 ), which resemble the cellular and anatomical composition of different regions of the human brain and can mimic disease pathways (Lancaster et al, 2013 ; Mariani et al, 2015 ; Garcez et al, 2016 ; Pollen et al, 2019 ; Velasco et al, 2019 ). Human brain organoids are becoming an emerging tool in TBI and mTBI as they provide a window into the injured tissues after trauma (Jgamadze et al, 2020 ; Ramirez et al, 2021a , b ) and over time into the chronic phase of injury (Silvosa et al, 2022 ).…”

Advances and Challenges in Cerebral Organoids Research

“…The cerebral organoid models of traumatic brain injury are amenable to scalability and time‐specific mechanistic investigation. [ 105 ]…”

A review of protocols for brain organoids and applications for disease modeling

“… Traumatic brain injury (TBI) Ramirez et al. 65 Model type Cerebral organoid Original publication Ramirez et al. 66 Method details Timeline Steps Fibroblast reprogramming to iPSCs 80 days Fibroblast culture Reprogramming iPSC characterization Generation of COs 50 days and up EB formation EB induction EB embedding Maturation Skull preparation 4–7 days Skull cleaning Skull craniotomy and sealing Gas sterilization Phantom brain preparation 20 min Gelatin and agarose mixture Skull filling with the phantom brain Controlled cortical impact (CCI) 2 h Stereotaxic frame sterilization Positioning COs CCI procedure CO recovery and analysis Phenotypes observed CCI induced apoptosis in COs and recapitulated pathological features of TBI, including neuronal damage, neuron loss, and astrogliosis.…”