A Micro-Optic Stalk (μOS) System to Model the Collective Migration of Retinal Neuroblasts

Contemporary regenerative therapies have introduced stem-like cells to replace damaged neurons in the visual system by recapitulating critical processes of eye development. The collective migration of neural stem cells is fundamental to retinogenesis and has been exceptionally well-studied using the fruit fly model of Drosophila Melanogaster. However, the migratory behavior of its retinal neuroblasts (RNBs) has been surprisingly understudied, despite being critical to retinal development in this invertebrate model. The current project developed a new microfluidic system to examine the collective migration of RNBs extracted from the developing visual system of Drosophila as a model for the collective motile processes of replacement neural stem cells. The system scales with the microstructure of the Drosophila optic stalk, which is a pre-cursor to the optic nerve, to produce signaling fields spatially comparable to in vivo RNB stimuli. Experiments used the micro-optic stalk system, or μOS, to demonstrate the preferred sizing and directional migration of collective, motile RNB groups in response to changes in exogenous concentrations of fibroblast growth factor (FGF), which is a key factor in development. Our data highlight the importance of cell-to-cell contacts in enabling cell cohesion during collective RNB migration and point to the unexplored synergy of invertebrate cell study and microfluidic platforms to advance regenerative strategies.

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A Perfused Microfluidic System to Study the Differentiation of Neural Stem Cells in vitro

Cells Tissues Organs ◽

10.1159/000495219 ◽

2018 ◽

Vol 206 (3) ◽

pp. 157-164 ◽

Cited By ~ 1

Author(s):

Qingxi Liu ◽

Sijie Xing ◽

Yupeng Liu ◽

Zijiang Zhang ◽

Lihui Lv ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Drug Evaluation ◽

Microfluidic System ◽

Quantitative Real Time Pcr ◽

Culture Systems ◽

Adult Neuron ◽

Cellular Microenvironments

Introduction: Due to the ability to mimic in vivo cellular microenvironments, the development of multicell culture systems has received increasing interest for use as research models and serving as platforms for drug evaluation. Methods: In this study, we developed a perfused microfluidic system to resemble the in vivo intercellular environment and applied it to study the differentiation from neural stem cells into neurons. Results: As determined by immunofluorescence chemistry and quantitative real-time PCR, the neural stem cells grown in this microfluidic system showed an elevated differentiation rate toward the formation of neurons as determined by the increased level of βIII-tubulin production, which is 4 times higher than that of culturing neural stem cells only. Conclusion: These results revealed that some factors secreted into the intercellular microenvironment by adult neuron cells can stimulate the differentiation of neural stem cells, pointing to the importance of developing multicellular culture systems such as the perfused microfluidic system we report here to better resemble the in vivo situation.

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Recapitulation of in vivo-like paracrine signals of human mesenchymal stem cells for functional neuronal differentiation of human neural stem cells in a 3D microfluidic system

Biomaterials ◽

10.1016/j.biomaterials.2015.06.011 ◽

2015 ◽

Vol 63 ◽

pp. 177-188 ◽

Cited By ~ 43

Author(s):

Kisuk Yang ◽

Hyun-Ji Park ◽

Sewoon Han ◽

Joan Lee ◽

Eunkyung Ko ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Neural Stem Cells ◽

Neuronal Differentiation ◽

Human Mesenchymal Stem Cells ◽

Microfluidic System ◽

Human Neural Stem Cells

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Faculty Opinions recommendation of In vivo analysis of quiescent adult neural stem cells responding to Sonic hedgehog.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1029162.342574 ◽

2005 ◽

Author(s):

Andrew Lumsden

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Sonic Hedgehog ◽

Adult Neural Stem Cells ◽

In Vivo Analysis

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Faculty Opinions recommendation of In vivo fate mapping and expression analysis reveals molecular hallmarks of prospectively isolated adult neural stem cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.7624956.7946061 ◽

2011 ◽

Author(s):

Anne Brunet ◽

Elena Mancini

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Expression Analysis ◽

Fate Mapping ◽

Adult Neural Stem Cells

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Regulated GDNF Delivery in Vivo Using Neural Stem Cells

10.21236/ada471929 ◽

2007 ◽

Author(s):

Clive Svendsen

Keyword(s):

Stem Cells ◽

Neural Stem Cells

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mRNA-Driven Generation of Transgene-Free Neural Stem Cells from Human Urine-Derived Cells

Cells ◽

10.3390/cells8091043 ◽

2019 ◽

Vol 8 (9) ◽

pp. 1043 ◽

Cited By ~ 1

Author(s):

Phil Jun Kang ◽

Daryeon Son ◽

Tae Hee Ko ◽

Wonjun Hong ◽

Wonjin Yun ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Human Urine ◽

Neurological Disorders ◽

Therapeutic Potential ◽

Embryonic Stem ◽

Global Gene Expression ◽

Patient Specific ◽

Human Neural Stem Cells

Human neural stem cells (NSCs) hold enormous promise for neurological disorders, typically requiring their expandable and differentiable properties for regeneration of damaged neural tissues. Despite the therapeutic potential of induced NSCs (iNSCs), a major challenge for clinical feasibility is the presence of integrated transgenes in the host genome, contributing to the risk for undesired genotoxicity and tumorigenesis. Here, we describe the advanced transgene-free generation of iNSCs from human urine-derived cells (HUCs) by combining a cocktail of defined small molecules with self-replicable mRNA delivery. The established iNSCs were completely transgene-free in their cytosol and genome and further resembled human embryonic stem cell-derived NSCs in the morphology, biological characteristics, global gene expression, and potential to differentiate into functional neurons, astrocytes, and oligodendrocytes. Moreover, iNSC colonies were observed within eight days under optimized conditions, and no teratomas formed in vivo, implying the absence of pluripotent cells. This study proposes an approach to generate transplantable iNSCs that can be broadly applied for neurological disorders in a safe, efficient, and patient-specific manner.

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Dynamic integration of enteric neural stem cells in ex vivo organotypic colon cultures

Scientific Reports ◽

10.1038/s41598-021-95434-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Georgina Navoly ◽

Conor J. McCann

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Organotypic Culture ◽

Ex Vivo ◽

Donor Cell ◽

Colonic Tissue ◽

Cell Therapies ◽

Enteric Ganglia ◽

Donor Cells

AbstractEnteric neural stem cells (ENSC) have been identified as a possible treatment for enteric neuropathies. After in vivo transplantation, ENSC and their derivatives have been shown to engraft within colonic tissue, migrate and populate endogenous ganglia, and functionally integrate with the enteric nervous system. However, the mechanisms underlying the integration of donor ENSC, in recipient tissues, remain unclear. Therefore, we aimed to examine ENSC integration using an adapted ex vivo organotypic culture system. Donor ENSC were obtained from Wnt1cre/+;R26RYFP/YFP mice allowing specific labelling, selection and fate-mapping of cells. YFP+ neurospheres were transplanted to C57BL6/J (6–8-week-old) colonic tissue and maintained in organotypic culture for up to 21 days. We analysed and quantified donor cell integration within recipient tissues at 7, 14 and 21 days, along with assessing the structural and molecular consequences of ENSC integration. We found that organotypically cultured tissues were well preserved up to 21-days in ex vivo culture, which allowed for assessment of donor cell integration after transplantation. Donor ENSC-derived cells integrated across the colonic wall in a dynamic fashion, across a three-week period. Following transplantation, donor cells displayed two integrative patterns; longitudinal migration and medial invasion which allowed donor cells to populate colonic tissue. Moreover, significant remodelling of the intestinal ECM and musculature occurred upon transplantation, to facilitate donor cell integration within endogenous enteric ganglia. These results provide critical evidence on the timescale and mechanisms, which regulate donor ENSC integration, within recipient gut tissue, which are important considerations in the future clinical translation of stem cell therapies for enteric disease.

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EXTH-07. TUMOR-HOMING INDUCED NEURAL STEM CELLS SECRETING A CYTOTOXIC PAYLOAD AS AN ADJUVANT TREATMENT FOR NON-SMALL CELL LUNG CANCER BRAIN METASTASES

Neuro-Oncology ◽

10.1093/neuonc/noaa215.361 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii88-ii88

Author(s):

Alison Mercer-Smith ◽

Wulin Jiang ◽

Alain Valdivia ◽

Juli Bago ◽

Scott Floyd ◽

...

Keyword(s):

Stem Cells ◽

Brain Metastases ◽

Neural Stem Cells ◽

Adjuvant Treatment ◽

Small Cell ◽

Small Cell Lung ◽

Tumor Homing ◽

Induced Neural Stem Cells

Abstract INTRODUCTION Non-small cell lung cancer (NSCLC) is the most common cancer to form brain metastases. Radiation treatment is standard-of-care, but recurrence is still observed in 40% of patients. An adjuvant treatment is desperately needed to track down and kill tumor remnants after radiation. Tumoritropic neural stem cells (NSCs) that can home to and deliver a cytotoxic payload offer potential as such an adjuvant treatment. Here we show the transdifferentiation of human fibroblasts into tumor-homing induced neural stem cells (hiNSCs) that secrete the cytotoxic protein TRAIL (hiNSC-TRAIL) and explore the use of hiNSC-TRAIL to treat NSCLC brain metastases. METHODS To determine the migratory capacity of hiNSCs, hiNSCs were infused intracerebroventricularly (ICV) into mice bearing established bilateral NSCLC H460 brain tumors. hiNSC accumulation at tumor foci was monitored using bioluminescent imaging and post-mortem fluorescent analysis. To determine synergistic effects of radiation with TRAIL on NSCLC, we performed in vitro co-culture assays and isobologram analysis. In vivo, efficacy was determined by tracking the progression and survival of mice bearing intracranial H460 treated with hiNSC-TRAIL alone or in combination with 2 Gy radiation. RESULTS/CONCLUSION Following ICV infusion, hiNSCs persisted in the brain for > 1 week and migrated from the ventricles to colocalize with bilateral tumor foci. In vitro, viability assays and isobologram analysis revealed the combination treatment of hiNSC-TRAIL and 2 Gy radiation induced synergistic killing (combination index=0.64). In vivo, hiNSC-TRAIL/radiation combination therapy reduced tumor volumes > 90% compared to control-treated animals while radiation-only and hiNSC-TRAIL-only treated mice showed 21% and 52% reduced volumes, respectively. Dual-treatment extended survival 40%, increasing survival from a median of 20 days in controls to 28 days in the treatment group. These results suggest hiNSC-TRAIL can improve radiation therapy for NSCLC brain metastases and could potentially improve outcomes for patients suffering from this aggressive form of cancer.

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