The Latest Articles on Neural Stem Cell Therapy

Our staff editors continue to share exciting, interesting, and thought-provoking reading material in the recommended articles series.

This week, we would like to share several latest articles on Neural Stem Cell Therapy.

Title: Analysis of autophagy deficiency and cytotoxicity in autophagy-deficient human embryonic stem cell-derived neurons
Authors: Miriam E. Korsgen, Congxin Sun, Elena Seranova, Malgorzata Zatyka, Dewi Astuti, Tetsushi Kataura, Timothy Barrett, Viktor I. Korolchuk, Sovan Sarkar
Type: Protocol
Abstract:
Autophagy, a catabolic process governing cellular and energy homeostasis, is essential for cell survival and human health. Here, we present a protocol for generating autophagy-deficient (ATG5-/-) human neurons from human embryonic stem cell (hESC)-derived neural precursors. We describe steps for analyzing loss of autophagy by immunoblotting. We then detail analysis of cell death by luminescence-based cytotoxicity assay and fluorescence-based TUNEL staining. This hESC-based experimental platform provides a genetic knockout model for undertaking autophagy studies relevant to human biology. For complete details on the use and execution of this protocol, please refer to Sun et al. (2023).1.
Access this article: https://doi.org/10.1016/j.xpro.2023.102529

Title: Survival and process outgrowth of human iPSC-derived cells expressing Purkinje cell markers in a mouse model for spinocerebellar degenerative disease
Authors: Takamasa Kamei, Atsushi Tamada, Toshiya Kimura, Akira Kakizuka, Akio Asai, Keiko Muguruma
Type: Research Articlet
Abstract:
Purkinje cells are the sole output neurons of the cerebellar cortex and play central roles in the integration of cerebellum-related motor coordination and memory. The loss or dysfunction of Purkinje cells due to cerebellar atrophy leads to severe ataxia. Here we used in vivo transplantation to examine the function of human iPS cell-derived cerebellar progenitors in adult transgenic mice in which Purkinje-specific cell death occurs due to cytotoxicity of polyglutamines. Transplantation using cerebellar organoids (42–48 days in culture), which are rich in neural progenitors, showed a viability of >50% 4 weeks after transplantation. STEM121+ grafted cells extended their processes toward the deep cerebellar nuclei, superior cerebellar peduncle, and vestibulocerebellar nuclei. The transplanted cells were mostly located in the white matter, and they were not found in the Purkinje cell layer. MAP2-positive fibers seen in the molecular layer of cerebellar cortex received VGluT2 inputs from climbing fibers. Transplanted neural progenitors overgrew in the host cerebellum but were suppressed by pretreatment with the γ-secretase inhibitor DAPT. Hyperproliferation was also suppressed by transplantation with more differentiated organoids (86 days in culture) or KIRREL2-positive cells purified by FACS sorting. Transplanted cells expressed Purkinje cell markers, GABA, CALB1 and L7, though they did not show fan-shaped morphology. We attempted to improve neuronal integration of stem cell-derived cerebellar progenitors by transplantation into the adult mouse, but this was not successfully achieved. Our findings in the present study contribute to regenerative medical application for cerebellar degeneration and provide new insights into cerebellar development in future.
Access this article: https://doi.org/10.1016/j.expneurol.2023.114511

Title: High-throughput assessment of metabolism-mediated neurotoxicity by combining 3D-cultured neural stem cells and liver cell spheroids
Authors: Pranav Joshi, Soo-Yeon Kang, Prabha Acharya, Darshita Sidhpura, Moo-Yeal Lee
Type: Research Article
Abstract:
Despite the fact that biotransformation in the liver plays an important role in the augmented toxicity and detoxification of chemicals, relatively little efforts have been made to incorporate biotransformation into in vitro neurotoxicity testing. Conventional in vitro systems for neurotoxicity tests lack the capability of investigating the qualitative and quantitative differences between parent chemicals and their metabolites in the human body. Therefore, there is a need for an in vitro toxicity screening system that can incorporate hepatic biotransformation of chemicals and predict the susceptibility of their metabolites to induce neurotoxicity. To address this need, we adopted 3D cultures of metabolically competent HepaRG cell line with ReNcell VM and established a high-throughput, metabolism-mediated neurotoxicity testing system. Briefly, spheroids of HepaRG cells were generated in an ultralow attachment (ULA) 384-well plate while 3D-cultured ReNcell VM was established on a 384-pillar plate with sidewalls and slits (384PillarPlate). Metabolically sensitive test compounds were added in the ULA 384-well plate with HepaRG spheroids and coupled with 3D-cultured ReNcell VM on the 384PillarPlate, which allowed us to generate metabolites in situ by HepaRG cells and test them against neural stem cells. We envision that this approach could be potentially adopted in pharmaceutical and chemical industries when high-throughput screening (HTS) is necessary to assess neurotoxicity of compounds and their metabolites.
Access this article: https://doi.org/10.1016/j.tiv.2023.105688

Title: Surface-tethered ROS-responsive micelle backpacks for boosting mesenchymal stem cell vitality and modulating inflammation in ischemic stroke treatment
Authors: Yang You, Yipu Liu, Chuchu Ma, Jianpei Xu, Laozhi Xie, Shiqiang Tong, Yinzhe Sun, Fenfen Ma, Yukun Huang, Junbin Liu, Wenze Xiao, Chengxiang Dai, Suke Li, Jigang Lei, Qiyong Mei, Xiaoling Gao, Jun Chen
Type: Research Article
Abstract:
Mesenchymal stem cells (MSCs) exhibited remarkable therapeutic potential in ischemic stroke due to their exceptional immunomodulatory ability and paracrine effect; they have also been regarded as excellent neuroprotectant delivery vehicles with inflammatory tropism. However, the presence of high levels of reactive oxygen species (ROS) and an oxidative stress environment at the lesion site inhibits cell survival and further therapeutic effects. Using bioorthogonal click chemistry, ROS-responsive luteolin-loaded micelles were tethered to the surface of MSCs. As MSCs migrated to the ischemic brain, the micelles would achieve ROS-responsive release of luteolin to protect MSCs from excessive oxidative damage while inhibiting neuroinflammation and scavenging ROS to ameliorate ischemic stroke. This study provided an effective and prospective therapeutic strategy for ischemic stroke and a framework for a stem cell-based therapeutic system to treat inflammatory cerebral diseases.
Access this article: https://doi.org/10.1016/j.jconrel.2023.08.039

Title: Designing biofunctional hydrogels for stem cell biology and regenerative medicine applications
Authors: Thai Thanh Hoang Thi, Maddison Laney, Hongmanlin Zhang, Fernando Martinez, Yunki Lee, Young C. Jang
Type: Review
Abstract:
Stem cell therapy has shown remarkable promise for treating various diseases and disorders. This strategy attenuates disease symptoms and stimulates endogenous and exogenous regeneration processes. However, several clinical trial shortcomings revealed that injecting only stem cells was insufficient to achieve satisfactory therapeutic benefits. This may be attributed to low cell survival, short-term cell retention, and limited functional improvements. To overcome this challenge, biofunctional hydrogels have been explored as a potential cell delivery platform and offer new possibilities for the next generation of stem cell-based therapies in clinics. Although the importance and impacts of hydrogels for stem cell delivery are clearly defined, this strategy should be carefully developed with all potential barriers in mind before moving to early-phase clinical trials. This review aims to provide a comprehensive overview of the design principles and applications of hydrogels for stem cell delivery in tissue engineering. We discuss the importance of creating cell-instructive hydrogels that mimic the three-dimensional (3D) stem cell niches and explore various hydrogel formulations employed in tissue engineering approaches. Finally, we review the remaining challenges of the current stem cell/hydrogel strategies in clinical trials and propose potential approaches for achieving successful clinical outcomes.
Access this article: https://doi.org/10.1016/j.jiec.2023.08.042