Human Model Research Articles

A de novo Missense Mutation in PPP2R5D Alters Dopamine Pathways and Morphology of iPSC-derived Midbrain Neurons.

Induced pluripotent stem cell (iPSC) models of neurodevelopmental disorders (NDDs) have promoted an understanding of commonalities and differences within or across patient populations by revealing the underlying molecular and cellular mechanisms contributing to disease pathology. Here, we focus on developing a human model for PPP2R5D-related NDD, called Jordan syndrome, which has been linked to Early-Onset Parkinson's Disease (EOPD). Here we sought to understand the underlying molecular and cellular phenotypes across multiple cell states and neuronal subtypes in order to gain insight into Jordan syndrome pathology. Our work revealed that iPSC-derived midbrain neurons from Jordan syndrome patients display significant differences in dopamine-associated pathways and neuronal architecture. We then evaluated a CRISPR-based approach for editing heterozygous dominant G-to-A mutations at the transcript level in patient-derived neural stem cells. Our findings show site-directed RNA editing is influenced by sgRNA length and cell type. These studies support the potential for a CRISPR RNA editor system to selectively edit mutant transcripts harboring G-to-A mutations in neural stem cells while providing an alternative editing technology for those suffering from NDDs.

Development of a novel computational technique to create DNA and cell geometrical models for Geant4-DNA

BackgroundThis study aimed to develop a novel human cell geometry for the Geant4-DNA simulation toolkit that explicitly incorporates all 23 chromosome pairs of the human cell. This approach contrasts with the existing, default human cell, geometrical model, which utilizes a continuous Hilbert curve. MethodsA Python-based tool named “complexDNA” was developed to facilitate the design of both simple and complex DNA geometries. This tool was employed to construct a human cell geometry with individual pairs of chromosomes. Subsequently, the performance of this chromosomal model was compared to the standard human cell model provided in the “molecularDNA” Geant4-DNA example. ResultsSimulations using the new chromosomal model revealed minimal discrepancies in DNA damage yield and fragment size distribution compared to the default human cell model. Notably, the chromosomal model demonstrated significant computational efficiency, requiring approximately three times less simulation time to achieve equivalent results. ConclusionsThis work highlights the importance of incorporating chromosomal structure into human cell models for radiation biology research. The “complexDNA” tool offers a valuable resource for creating intricate DNA structures for future studies. Further refinements, such as implementing smaller voxels for euchromatin regions, are proposed to enhance the model’s accuracy.

Standalone single- and bi-layered human skin 3D models supported by recombinant silk feature native spatial organization.

Physiologically relevant human skin models that include key skin cell types can be used for in vitro drug testing, skin pathology studies, or clinical applications such as skin grafts. However, there is still no golden standard for such a model. We investigated the potential of a recombinant function-alized spider silk protein, FN-silk, for the construction of a dermal, an epidermal, and a bilayered skin equivalent (BSE). Specifically, two formats of FN-silk (i.e., 3D network and nanomembrane) were evaluated. The 3D network was used as an elastic ECM-like support for the dermis, and the thin, permeable nanomembrane was used as a basement membrane to support the epidermal epi-thelium. Immunofluorescence microscopy and spatially resolved transcriptomics analysis demon-strated the secretion of key ECM components and the formation of microvascular-like structures. Furthermore, the epidermal layer exhibited clear stratification and the formation of a cornified layer, resulting in a tight physiologic epithelial barrier. Our findings indicate that the presented FN-silk-based skin models can be proposed as physiologically relevant standalone epidermal or dermal models, as well as a combined BSE.

Investigating Inherited Heart Diseases Using Human Induced Pluripotent Stem Cell-Based Models

Inherited heart diseases (IHDs) are caused by genetic mutations that disrupt the physiological structure and function of the heart. Understanding the mechanisms behind these diseases is crucial for developing personalised interventions in cardiovascular medicine. Development of induced pluripotent stem cells, which can then be differentiated to any nucleated adult cell type, has enabled the creation of personalised single-cell and multicellular models, providing unprecedented insights into the pathophysiology of IHDs. This review provides a comprehensive overview of recent advancements in human iPSC models used to dissect the molecular and genetic underpinnings of common IHDs. We examine multicellular models and tissue engineering approaches, such as cardiac organoids, engineered heart tissue, and multicellular co-culture systems, which simulate complex intercellular interactions within heart tissue. Recent advancements in stem cell models offer a more physiologically relevant platform to study disease mechanisms, enabling researchers to observe cellular interactions, study disease progression, and identify therapeutic strategies. By leveraging these innovative models, we can gain deeper insights into the molecular and cellular mechanisms underlying IHDs, ultimately paving the way for more effective diagnostic and therapeutic strategies.

CCL2 blockade combined with PD-1/P-selectin immunomodulators impedes breast cancer brain metastasis.

Over the last two decades, the diagnosis and treatment of breast cancer patients have considerably improved. However, brain metastases remain a major clinical challenge and a leading cause of mortality. Thus, a better understanding of the pathways involved in the metastatic cascade is essential. To this end, we have investigated the reciprocal effects of astrocytes and breast cancer cells, employing traditional 2-dimensional cell culture and our unique 3-dimensional multicellular tumoroid models. Our findings revealed that astrocytes enhance the proliferation, migration, and invasion of breast cancer cells, suggesting a supportive role for astrocytes in breast cancer outgrowth to the brain. Elucidating the key players in astrocyte-breast cancer cells crosstalk, we found that CCL2 is highly expressed in breast cancer brain metastases tissue sections from both patients and mice. Our in vitro and in vivo models further confirmed that CCL2 has a functional role in brain metastasis. Given their aggressive nature, we sought additional immune checkpoints for rationale combination therapy. Among the promising candidates were the adhesion molecule P-selectin, which we have recently shown to play a key role in the crosstalk with microglia cells, and the co-inhibitory receptor PD-1, the main target of currently approved immunotherapies. Finally, combining CCL2 inhibition with immunomodulators targeting either PD-1/PD-L1 or P-selectin/P-Selectin Ligand-1 axes in our human 3-dimensional tumoroid models and in vivo presented more favorable outcomes than each monotherapy. Taken together, we propose that CCL2-CCR2/CCR4 is a key pathway promoting breast cancer brain metastases and a promising target for an immunotherapeutic combination approach.

Development and Characterization of a Human Mammary Epithelial Cell Culture Model for the Blood–Milk Barrier—A Contribution from the ConcePTION Project

It is currently impossible to perform an evidence-based risk assessment for medication use during breastfeeding. The ConcePTION project aims to provide information about the use of medicines during lactation. The study aimed to develop and characterize an in vitro model of the blood–milk barrier to determine the extent of the milk transfer of xenobiotics, relying on either on human mammary epithelial cells (hMECs) or immortalized cell lines derived from breast tissue. The hMECs were cultured and characterized for epithelial markers; further, the ability to form an epithelial barrier was investigated. Drug transporter functionality in the cultured hMECs was analyzed with specific probe substrates. The hMECs showed an epithelial morphology and the expression of epithelial markers and tight junctions. They formed a reproducible tight barrier with a transepithelial electrical resistance greater than 400 Ωcm2, unlike immortalized cell lines. Different levels of mRNA expression were detected for 81 genes of membrane transporters. Functional assays showed no evidence for the transporter-mediated secretion of medicines across the hMECs. Nevertheless, the hMEC-based in vitro model covered a 50-fold range of permeability values, differentiating between passive transcellular and paracellular-mediated transport. The cultured hMECs proved to be a promising in vitro model for biorelevance; the wide characterization of hMECs makes them useful for studying medicine partitioning in milk.

3D bioprinted in vitro epilepsy models for pharmacological evaluation in temporal lobe epilepsy.

This study introduces a novel in vitro model for intractable temporal lobe epilepsy (TLE) utilizing 3D bioprinting technology, aiming to replicate the complex neurobiological characteristics of TLE more accurately . Primary neural cell constructs were fabricated and subjected to epileptiform-inducing conditions, fostering synaptic proliferation and neuronal loss. Systematically electrophysiological and immunofluorescent analyses indicated that significant synaptic connectivity and sustained epileptiform activities within the constructs akin to those observed in human epilepsy models. Notably, the model responded to treatments with phenytoin and tetrodotoxin, illustrating its potential utility in drug response kinetics studies. Furthermore, we performed drug permeability simulations using COMSOL Multiphysics to analyze the diffusion characteristics of these drugs within the constructs. These results confirm that our 3D bioprinted neural model provides a physiologically relevant and ethically sustainable platform, which is beneficial for studying TLE mechanisms and developing therapeutic strategies with high accuracy and clinical relevance.&#xD.

Electromagnetic Compatibility Study of Trackside Antenna Array Miniaturization in the Subway Tunnel

Abstract To improve the compatibility of the subway tunnel’s electromagnetic environment and reduce the radiation impact of trackside antennas on subway workers. This paper proposes a miniaturized dual-band trackside antenna array by using metamaterial units. Its operation bandwidth is 2.33~2.56 GHz and 3.24~3.45 GHz, which could simultaneously satisfy the signal cover demands of the communications-based train control (CBTC) and the civil 5G wireless communication system. The proposed miniaturized antenna array has a maximum gain of 14.4 dBi and a maximum channel capacity of 13.9 bps/Hz at a signal-to-noise ratio (SNR) of 20 dB, which can effectively improve the quality of wireless communication systems. The number of trackside antennas with a single operation frequency band is reduced, and the distance between the antennas is enlarged at the same time. Besides that, we analyze the radiation impact on the tunnel electromagnetic environment of the proposed trackside antenna array. In particular, the electromagnetic dose absorbed by the human model of a tunnel worker is quantized. The results show that the electric field strength in the tunnel reduces by 4.11% at least after antenna array miniaturization, and the specific absorption rate (SAR) absorbed by the worker model's trunk, skull, brain, heart, and liver is reduced by a maximum of 19.02%, 33.16%, 28.27%, 41.75%, and 74.54%, respectively, further lowering the human electromagnetic exposure risk. Therefore, a miniaturized trackside antenna array could reduce the interference from other radiation sources in the tunnel while providing a new idea for electromagnetic protection for subway workers.

Evaluation of resurgence following differential reinforcement of alternative behavior with and without extinction in a human operant model.

One of the most common treatments for severe challenging behavior involves placing the challenging behavior on extinction and differentially reinforcing an alternative response (DRA). However, extinction is not always feasible and may be unsafe or impractical to implement in some circumstances. Thus, implementing a DRA without extinction intervention may be necessary for some cases. Currently, the extent to which DRA without extinction produces durable treatment outcomes, particularly as it relates to the resurgence of challenging behavior, is unclear. The present study investigated resurgence following DRA with and without extinction using a three-phase resurgence evaluation in a translational human operant model with college students as participants. All participants demonstrated resurgence across both experimental groups. Although there were no statistically significant differences in the prevalence, magnitude, or persistence of resurgence between groups, levels of resurgence magnitude were relatively higher in the DRA-without-extinction group than in the DRA-with-extinction group. Clinical implications of these findings and directions for future human operant investigations of resurgence are discussed.

Advances in bacterial artificial chromosome (BAC) transgenic mice for gene analysis and disease research.

Transgenic mice, including those created using Bacterial Artificial Chromosomes (BACs), are artificial manipulations that have become critical tools for studying gene function. While conventional transgenic techniques face challenges in achieving precise expression of foreign genes in specific cells and tissues, BAC transgenic mice offer a solution by incorporating large DNA segments that can include entire expression units with tissue-specific enhancers. This review provides a thorough examination of BAC transgenic mouse technology, encompassing both traditional and humanized models. We explore the benefits and drawbacks of BAC transgenesis compared to other techniques such as knock-in and CRISPR/Cas9 technologies. The review emphasizes the applications of BAC transgenic mice in various disciplines, including neuroscience, immunology, drug metabolism, and disease modeling. Additionally, we address crucial aspects of generating and analyzing BAC transgenic mice, such as position effects, copy number variations, and strategies to mitigate these challenges. Despite certain limitations, humanized BAC transgenic mice have proven to be invaluable tools for studying the pathogenesis of human diseases, drug development, and understanding intricate gene regulatory mechanisms. This review discusses current topics on BAC transgenic mice and their evolving significance in biomedical research.

Vagus nerve stimulation in Parkinson’s disease: a scoping review of animal studies and human subjects research

Parkinson’s Disease (PD) is a prevalent, progressive neurodegenerative disease with motor and non-motor symptoms. Vagus Nerve Stimulation (VNS) has emerged as a potential therapeutic approach for PD, but published research on this topic varies widely. This scoping review maps existing literature on VNS for PD, highlighting stimulation methods, operational parameters, safety profiles, neurophysiological mechanisms, and clinical outcomes in human and animal models. Online databases were used to identify 788 papers published between 2013 and 2023, from which 17 publications on invasive and non-invasive VNS in PD were selected. Studies showed high variability in VNS parameters and study design. Evidence in animal models and human subjects suggests potential neurophysiological effects on PD-related pathology and motor function improvements. However, significant gaps in the literature remain. Future research should include rigorous reporting of study design, standardization of stimulation parameters, and larger sample sizes to ultimately facilitate translation of VNS into clinical practice.

RBBP6-Mediated ERRα Degradation Contributes to Mitochondrial Injury in Renal Tubular Cells in Diabetic Kidney Disease.

Diabetic Kidney Disease (DKD), a major precursor to end-stage renal disease, involves mitochondrial dysfunction in proximal renal tubular cells (PTCs), contributing to its pathogenesis. Estrogen-related receptor α (ERRα) is essential for mitochondrial integrity in PTCs, yet its regulation in DKD is poorly understood. This study investigates ERRα expression and its regulatory mechanisms in DKD, assessing its therapeutic potential. Using genetic, biochemical, and cellular approaches, ERRα expression Was examined in human DKD specimens and DKD mouse models. We identified the E3 ubiquitin ligase retinoblastoma binding protein 6 (RBBP6) as a regulator of ERRα, promoting its degradation through K48-linked polyubiquitination at the K100 residue. This degradation pathway significantly contributed to mitochondrial injury in PTCs of DKD models. Notably, conditional ERRα overexpression or RBBP6 inhibition markedly reduced mitochondrial damage in diabetic mice, highlighting ERRα's protective role in maintaining mitochondrial integrity. The interaction between RBBP6 and ERRα opens new therapeutic avenues, suggesting that modulating RBBP6-ERRα interactions could be a strategy for preserving mitochondrial function and slowing DKD progression.

Comparative analysis of mechanical and drilling properties: Human skull vs. 3D-printed replicas for neurosurgical training

Neurosurgery is one of the most difficult surgical disciplines, making neurosurgical simulation crucial for learners. 3D printing is increasingly used for this simulation due to its accessibility and cost-effectiveness compared to conventional methods. Previous studies have qualitatively evaluated the efficacy of 3D printing models for burr hole and craniotomy simulation using surgeon feedback. This study quantitatively evaluates the mechanical properties of human skulls and compares them with 3D-printed skulls to identify the necessary materials and technologies for accurate replication in neurosurgical training. Vickers hardness, compression, and drilling simulation tests were conducted on human cadaveric skulls and 3D-printed models manufactured using Fused Filament Fabrication (FFF), Stereolithography (SLA), and Material Jetting (MJ) technologies. Uniaxial force during a simulated burr hole procedure was measured as drilling progressed through the outer table (OT), diploë and inner table (IT) of the skull. The results showed that different 3D printing technologies and materials have qualities corresponding to each of the three layers of the human skull. For instance, the White Resin 40 model was the closest match to the OT of human skull, exhibiting the lowest mean difference of drilling modulus of 1.98. The OT of the human skull had a Vickers hardness of 38.6 ± 5 HV0.05. The closest 3D-printed models were SLA White at 16.6 ± 0.3 HV0.05 and Rigid 10K at 65.7 ± 3 HV0.05. In terms of mechanical properties, the human skull demonstrated a compression modulus that closely matched the SkullSTN 2, PC 60, PEEK 40, and PEEK 60, with differences of less than 0.1 GPa. These findings demonstrate the effectiveness of drilling simulation tests in evaluating various materials and infills to refine models before neurosurgeon evaluation, enabling the identification of an optimal 3D-printed training model for simulating burr hole procedures.

Hinotori™ robotic esophagectomy: a feasibility cadaver study.

This preclinical feasibility study investigates the potential of utilizing the hinotori™ robot system for esophagectomy. In three human cadaver models, the esophagus was successfully mobilized and resected using the hinotori™ system, with a mean thoracic procedure time of 57minutes. The system allowed for precise dissection and radical lymphadenectomy without arm collision, attributed to its versatile design and docking-free trocars. Standard robot-specific patient positioning, including a 35° left lateral inclination, and trocar placement in a posterior axillary line configuration were employed. Notably, trocars suitable for both laparoscopy and the hinotori™ robot were utilized, providing flexibility in trocar selection. Unique features, such as the ergonomic console and pointer-based pivot point identification system, contributed to procedural success. While these findings highlight the promising potential of the hinotori™ system in advancing esophageal surgery, further clinical studies are warranted to validate its reproducibility and clinical utility. Additionally, enhancements to the pivot point identification system and evaluation of the arm base's features may further optimize surgical outcomes.

Hemozoin induces malaria via activation of DNA damage, p38 MAPK and neurodegenerative pathways in a human iPSC-derived neuronal model of cerebral malaria

Malaria caused by Plasmodium falciparum infection results in severe complications including cerebral malaria (CM), in which approximately 30% of patients end up with neurological sequelae. Sparse in vitro cell culture-based experimental models which recapitulate the molecular basis of CM in humans has impeded progress in our understanding of its etiology. This study employed healthy human induced pluripotent stem cells (iPSCs)-derived neuronal cultures stimulated with hemozoin (HMZ) - the malarial toxin as a model for CM. Secretome, qRT-PCR, Metascape, and KEGG pathway analyses were conducted to assess elevated proteins, genes, and pathways. Neuronal cultures treated with HMZ showed enhanced secretion of interferon-gamma (IFN-γ), interleukin (IL)1-beta (IL-1β), IL-8 and IL-16. Enrichment analysis revealed malaria, positive regulation of cytokine production and positive regulation of mitogen-activated protein kinase (MAPK) cascade which confirm inflammatory response to HMZ exposure. KEGG assessment revealed up-regulation of malaria, MAPK and neurodegenerative diseases-associated pathways which corroborates findings from previous studies. Additionally, HMZ induced DNA damage in neurons. This study has unveiled that exposure of neuronal cultures to HMZ, activates molecules and pathways similar to those observed in CM and neurodegenerative diseases. Furthermore, our model is an alternative to rodent experimental models of CM.

Establishment of a Serum-Free Human iPSC-Derived Model of Peripheral Myelination.

Myelination and the formation of nodes of Ranvier are essential for the rapid conduction of nerve impulses along axons in the peripheral nervous system (PNS). While many animal-based and serum-containing models of peripheral myelination have been developed, these have limited ability when it comes to studying genetic disorders affecting peripheral myelination. We report a fully induced pluripotent stem cell (iPSC)-derived human model of peripheral myelination using Schwann cells (SCs) and motoneurons, cultured in a serum-free medium on patterned and nonpatterned surfaces. Results demonstrated iPSC-derived SC-expressed early growth response protein 2 (Egr2), a key transcription factor for myelination, and after ∼30 days in coculture, hallmark features of myelination, including myelin segment and node of Ranvier formation, were observed. Myelin segments were stained for the myelin basic protein, which surrounded neurofilament-stained motoneuron axons. Clusters of voltage-gated sodium channels flanked by paranodal protein contactin-associated protein 1, indicating node of Ranvier formation, were also observed. High-resolution confocal microscopy allowed for 3D reconstruction and measurement of myelin g-ratios of myelin segments, with an average g-ratio of 0.67, consistent with reported values in the literature, indicating mature myelin segment formation. This iPSC-based model of peripheral myelination provides a platform to investigate numerous PNS diseases, including Charcot-Marie Tooth disorder, Guillian-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, and antimyelin-associated glycoprotein peripheral neuropathy, with the potential for greater translatability to humans for improving the applicability for drug-screening programs.

Multi-drug resistant Staphylococcus epidermidis from chronic wounds impair healing in human wound model.

Venous leg ulcers (VLUs) represent one of the most prevalent types of chronic wounds characterised by perturbed microbiome and biofilm-forming bacteria. As one of the most abundant skin-commensal, Staphylococcus epidermidis is known as beneficial for the host, however, some strains can form biofilms and hinder wound healing. In this study, S. epidermidis distribution in VLUs and associated resistome were analysed in ulcer tissue from patients. Virulence of S. epidermidis isolates from VLUs were evaluated by whole genome sequencing, antimicrobial susceptibility testing, in vitro biofilm and binding assays, and assessment of biofilm-forming capability and pro-inflammatory potential using human ex vivo wound model. We demonstrated that S. epidermidis isolates from VLUs inhibit re-epithelialization through biofilm-dependent induction of IL-1β, IL-8, and IL-6 which was in accordance with impaired healing outcomes observed in patients. High extracellular matrix binding ability of VLU isolates was associated with antimicrobial resistance and expression levels of the embp and sdrG, responsible for bacterial binding to fibrinogen and fibrin, respectively. Finally, we showed that S. epidermidis from VLUs demonstrate pathogenic features with ability to impair healing which underscores the emergence of treatment-resistant virulent lineages in patients with chronic ulcers.

AMG 193, a Clinical Stage MTA-Cooperative PRMT5 Inhibitor, Drives Antitumor Activity Preclinically and in Patients With MTAP-Deleted Cancers.

One of the most robust synthetic lethal interactions observed in multiple functional genomic screens has been dependency on PRMT5 in cancer cells with MTAP deletion. We report the discovery of the clinical stage MTA-cooperative PRMT5 inhibitor AMG 193, which preferentially binds PRMT5 in the presence of MTA and has potent biochemical and cellular activity in MTAP-deleted cells across multiple cancer lineages. In vitro, PRMT5 inhibition induces DNA damage, cell cycle arrest, and aberrant alternative mRNA splicing in MTAP-deleted cells. In human cell line and patient-derived xenograft models, AMG 193 induces robust antitumor activity and is well tolerated with no impact on normal hematopoietic cell lineages. AMG 193 synergizes with chemotherapies or the KRAS G12C inhibitor sotorasib in vitro, and combination treatment in vivo significantly inhibits tumor growth. AMG 193 is demonstrating promising clinical activity, including confirmed partial responses in patients with MTAP-deleted solid tumors from an ongoing phase 1/2 study.

Ginsenoside Rk3 Treats Corneal Injury Through the HMGB1/TLR4/NF-κB Pathway.

The cornea serves as a vital protective shield for the eye, safeguarding its intricate internal structures from external threats. Damage to the cornea compromises this protective function, triggering inflammation and potentially causing long-term harm. While ginsenoside Rk3 has demonstrated potential for repairing the corneal barrier and reducing inflammation, its effectiveness in treating corneal damage remains relatively unexplored. This comprehensive study uses both in vivo and in vitro models to investigate the therapeutic capabilities of ginsenoside Rk3. Using two models of corneal damage, a benzalkonium chloride-induced mouse model and a high osmolarity-induced human corneal epithelial cell model, we scrutinized the effects of ginsenoside Rk3 treatment. Our results showed that ginsenoside Rk3-treated mice manifested reduced corneal damage and inflammation compared with their untreated counterparts. Furthermore, mice treated with ginsenoside Rk3 exhibited an organized arrangement of corneal cells and diminished stromal layer thickness, indicating reparative properties of ginsenoside Rk3. Additionally, ginsenoside Rk3 increased the expression of tight junction proteins, suppressed inflammatory factors, and decreased HMGB1 protein expression, thereby modulating downstream signaling pathways. Collectively, our findings present compelling evidence that ginsenoside Rk3 is a promising therapeutic option for corneal injury. By repairing the corneal barrier, mitigating inflammation, and modulating specific protein levels, ginsenoside Rk3 opens new avenues for managing corneal damage.

Home-based guidance training system with interactive visual feedback using kinect on stroke survivors with moderate to severe motor impairment

The home-based training approach benefits stroke survivors by providing them with an increased amount of training time and greater feasibility in terms of their training schedule, particularly for those with severe motor impairment. Computer-guided training systems provide visual feedback with correct movement patterns during home-based training. This study aimed to investigate the improvement in motor performance among stroke survivors with moderate to severe motor impairment after 800 min of training using a home-based guidance training system with interactive visual feedback. Twelve patients with moderate to severe stroke underwent home-based training, totaling 800 min (20–40 min per session, with a frequency of 3 sessions per week). The home-based guidance training system uses Kinect to reconstruct the 3D human body skeletal model and provides real-time motor feedback during training. The training exercises consisted of six core exercises and eleven optional exercises, including joint exercises, balance control, and coordination. Pre-training and post-training assessments were conducted using the Fugl-Meyer Assessment-Upper Limb (FMA-UE), Fugl-Meyer Assessment-Lower Limb (FMA-LE), Functional Ambulation Categories (FAC), Berg Balance Scale (BBS), Barthel Index (BI), Modified Ashworth Scale (MAS), as well as kinematic data of joint angles and center of mass (COM). The results indicated that motor training led to the attainment of the upper limit of functional range of motion (FROM) in hip abduction, shoulder flexion, and shoulder abduction. However, there was no improvement in the active range of motion (AROM) in the upper extremity (U/E) and lower extremity (L/E) joints, reaching the level of the older healthy population. Significant improvements were observed in both left/right and superior/inferior displacements, as well as body sway in the mediolateral axis of the COM, after 800 min of training. In conclusion, the home-based guidance system using Kinect aids in improving joint kinematics performance at the level of FROM and balance control, accompanied by increased mediolateral body sway of the COM for stroke survivors with moderate to severe stroke. Additionally, spasticity was reduced in both the upper and lower extremities after 800 min of home-based training.