Urine Sample-Derived Cerebral Organoids Suitable for Studying Neurodevelopment and Pharmacological Responses

Lin, Victor; Hu, Jiangnan; Zolekar, Ashwini; Lv, Yan; Wang, Yu-Chieh

doi:10.3389/fcell.2020.00304

Cited by 11 publications

(10 citation statements)

References 49 publications

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“…During cerebral development, SHH promotes the hindbrain and posterior forebrain identity [ 53 , 55 ]. However, cyclopamine leads to a near complete inhibition of SHH signaling activity [ 53 ], and contributes to anterior characteristics, such as the expression of FOXG1 and Nkx2.1 [ 56 , 57 ], as examined in our study. However, applying 3D brain organoids in vitro culture systems to assess the therapeutic potential of the secreted EVs has not been well investigated [ 18 , 19 ].…”

Section: Discussionmentioning

confidence: 94%

Human Forebrain Organoid-Derived Extracellular Vesicle Labeling with Iron Oxides for In Vitro Magnetic Resonance Imaging

et al. 2022

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The significant roles of extracellular vesicles (EVs) as intracellular mediators, disease biomarkers, and therapeutic agents, make them a scientific hotspot. In particular, EVs secreted by human stem cells show significance in treating neurological disorders, such as Alzheimer’s disease and ischemic stroke. However, the clinical applications of EVs are limited due to their poor targeting capabilities and low therapeutic efficacies after intravenous administration. Superparamagnetic iron oxide (SPIO) nanoparticles are biocompatible and have been shown to improve the targeting ability of EVs. In particular, ultrasmall SPIO (USPIO, <50 nm) are more suitable for labeling nanoscale EVs due to their small size. In this study, induced forebrain neural progenitor cortical organoids (iNPCo) were differentiated from human induced pluripotent stem cells (iPSCs), and the iNPCo expressed FOXG1, Nkx2.1, α-catenin, as well as β-tubulin III. EVs were isolated from iNPCo media, then loaded with USPIOs by sonication. Size and concentration of EV particles were measured by nanoparticle tracking analysis, and no significant changes were observed in size distribution before and after sonication, but the concentration decreased after labeling. miR-21 and miR-133b decreased after sonication. Magnetic resonance imaging (MRI) demonstrated contrast visualized for the USPIO labeled EVs embedded in agarose gel phantoms. Upon calculation, USPIO labeled EVs exhibited considerably shorter relaxation times, quantified as T2 and T2* values, reducing the signal intensity and generating higher MRI contrast compared to unlabeled EVs and gel only. Our study demonstrated that USPIO labeling was a feasible approach for in vitro tracking of brain organoid-derived EVs, which paves the way for further in vivo examination.

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Section: Discussionmentioning

confidence: 94%

Human Forebrain Organoid-Derived Extracellular Vesicle Labeling with Iron Oxides for In Vitro Magnetic Resonance Imaging

et al. 2022

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“…These debilitating conditions impede in many cases their participation in clinical trials, which is a major challenge for the translation of pediatric cancer research into clinical practice. Unlike solid biopsies, urine collection represents an easy, safe, pain-free and inexpensive way of obtaining large quantities of somatic cells to engineer cell-based therapeutics for a variety of diseases ( 11 , 36 , 37 ). Here we report for the first time the generation of two iPSC lines from UDCs of children with brain tumors.…”

Section: Discussionmentioning

confidence: 99%

Generation of mesenchymal stromal cells from urine-derived iPSCs of pediatric brain tumor patients

et al. 2023

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Human induced pluripotent stem cells (iPSCs) provide a virtually inexhaustible source of starting material for next generation cell therapies, offering new opportunities for regenerative medicine. Among different cell sources for the generation of iPSCs, urine cells are clinically relevant since these cells can be repeatedly obtained by non-invasive methods from patients of any age and health condition. These attributes encourage patients to participate in preclinical and clinical research. In particular, the use of urine-derived iPSC products is a convenient strategy for children with brain tumors, which are medically fragile patients. Here, we investigate the feasibility of using urine samples as a source of somatic cells to generate iPSC lines from pediatric patients with brain tumors (BT-iPSC). Urinary epithelial cells were isolated and reprogrammed using non-integrative Sendai virus vectors harboring the Yamanaka factors KLF4, OCT3/4, SOX2 and C-MYC. After reprogramming, BT-iPSC lines were subject to quality assessment and were compared to iPSCs obtained from urine samples of non-tumor pediatric patients (nonT-iPSC). We demonstrated that iPSCs can be successfully derived from a small volume of urine obtained from pediatric patients. Importantly, we showed that BT-iPSCs are equivalent to nonT-iPSCs in terms of morphology, pluripotency, and differentiation capacity into the three germ layers. In addition, both BT-iPSCs and nonT-iPSCs efficiently differentiated into functional mesenchymal stem/stromal cells (iMSC) with immunomodulatory properties. Therefore, this study provides an attractive approach to non-invasively generate personalized iMSC products intended for the treatment of children with brain tumors.

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“…Our search of the literature identified several manuscripts that used a modified version of the Lancaster method, although no acknowledgement to Lancaster et al was made. We also found a manuscript that generated organoids from hiPSC derived from urine epithelial cells, which did not acknowledge Stemcell Technologies, who published this method of hiPSC generation in 2018 [86,87]. Several viral pathogenesis studies have been published using the unguided protocol, most likely due to the reproducibility of the Lancaster method and the numerous modifications available to it (Table 2) [25].…”

Section: Unguided Organoid Modelsmentioning

confidence: 99%

Methodologies for Generating Brain Organoids to Model Viral Pathogenesis in the CNS

2021

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(1) Background: The human brain is of interest in viral research because it is often the target of viruses. Neurological infections can result in consequences in the CNS, which can result in death or lifelong sequelae. Organoids modeling the CNS are notable because they are derived from stem cells that differentiate into specific brain cells such as neural progenitors, neurons, astrocytes, and glial cells. Numerous protocols have been developed for the generation of CNS organoids, and our goal was to describe the various CNS organoid models available for viral pathogenesis research to serve as a guide to determine which protocol might be appropriate based on research goal, timeframe, and budget. (2) Methods: Articles for this review were found in Pubmed, Scopus and EMBASE. The search terms used were “brain + organoid” and “CNS + organoid” (3) Results: There are two main methods for organoid generation, and the length of time for organoid generation varied from 28 days to over 2 months. The costs for generating a population of organoids ranged from USD 1000 to 5000. (4) Conclusions: There are numerous methods for generating organoids representing multiple regions of the brain, with several types of modifications for fine-tuning the model to a researcher’s specifications. Organoid models of the CNS can serve as a platform for characterization and mechanistic studies that can reduce or eliminate the use of animals, especially for viruses that only cause disease in the human CNS.

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Urine Sample-Derived Cerebral Organoids Suitable for Studying Neurodevelopment and Pharmacological Responses

Cited by 11 publications

References 49 publications

Human Forebrain Organoid-Derived Extracellular Vesicle Labeling with Iron Oxides for In Vitro Magnetic Resonance Imaging

Human Forebrain Organoid-Derived Extracellular Vesicle Labeling with Iron Oxides for In Vitro Magnetic Resonance Imaging

Generation of mesenchymal stromal cells from urine-derived iPSCs of pediatric brain tumor patients

Methodologies for Generating Brain Organoids to Model Viral Pathogenesis in the CNS

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