| 1. |
|
|
| 2. |
- Frischknecht, R., et al.
(author)
-
Comparison of the environmental assessment of an identical office building with national methods
- 2019
-
In: IOP Conference Series: Earth and Environmental Science. - : IOP Publishing. - 1755-1307 .- 1755-1315. ; , s. 012037-
-
Conference paper (peer-reviewed)abstract
- The IEA EBC Annex 72 focuses on the assessment of the primary energy demand, greenhouse gas emissions and environmental impacts of buildings during production, construction, use (including repair and replacement) and end of life (dismantling), i.e. during the entire life cycle of buildings. In one of its activities, reference buildings (size, materialisation, operational energy demand, etc.) were defined on which the existing national assessment methods are applied using national (if available) databases and (national/regional) approaches. The "be2226" office building in Lustenau, Austria was selected as one of the reference buildings. TU Graz established a BIM model and quantified the amount of building elements as well as construction materials required and the operational energy demand. The building assessment was carried out using the same material and energy demand but applying the LCA approach used in the different countries represented by the participating Annex experts. The results of these assessments are compared in view of identifying major discrepancies. Preliminary findings show that the greenhouse gas emissions per kg of building material differ up to a factor of two and more. Major differences in the building assessments are observed in the transports to the construction site (imports) and the construction activities as well as in the greenhouse gas emissions of the operational energy demand (electricity). The experts document their practical difficulties and how they overcame them. The results of this activity are used to better target harmonisation efforts.
|
|
| 3. |
- Frischknecht, R., et al.
(author)
-
Comparison of the greenhouse gas emissions of a high-rise residential building assessed with different national LCA approaches - IEA EBC Annex 72
- 2020
-
In: IOP Conference Series. - : IOP Publishing. ; , s. 022029-
-
Conference paper (peer-reviewed)abstract
- Introduction: The international research project IEA EBC Annex 72 investigates the life cycle related environmental impacts caused by buildings. The project aims inter alia to harmonise LCA approaches on buildings. Methods: To identify major commonalities and discrepancies among national LCA approaches, reference buildings were defined to present and compare the national approaches. A residential high-rise building located in Tianjin, China, was selected as one of the reference buildings. The main construction elements are reinforced concrete shear walls, beams and floor slabs. The building has an energy reference area of 4566 m2 and an operational heating energy demand of 250 MJ/m2a. An expert team provided information on the quantities of building materials and elements required for the construction, established a BIM model and quantified the operational energy demand. Results: The greenhouse gas emissions and environmental impacts of the building were quantified using 17 country-specific national assessment methods and LCA databases. Comparisons of the results are shown on the level of building elements as well as the complete life cycle of the building. Conclusions: The results of these assessments show that the main differences lie in the LCA background data used, the scope of the assessment and the reference study period applied. Despite the variability in the greenhouse gas emissions determined with the 17 national methods, the individual results are relevant in the respective national context of the method, data, tool and benchmark used. It is important that environmental benchmarks correspond to the particular LCA approach and database of a country in which the benchmark is applied. Furthermore, the results imply to include building technologies as their contribution to the overall environmental impacts is not negligible. Grant support: The authors thank the IEA for its organizational support and the funding organizations in the participating countries for their financial support.
|
|
| 4. |
- Pircs, Karolina, et al.
(author)
-
Distinct subcellular autophagy impairments in induced neurons from patients with Huntington's disease
- 2022
-
In: Brain : a journal of neurology. - : Oxford University Press (OUP). - 1460-2156. ; 145:9, s. 3035-3057
-
Journal article (peer-reviewed)abstract
- Huntington's disease (HD) is a neurodegenerative disorder caused by CAG expansions in the huntingtin (HTT) gene. Modelling Huntington's disease is challenging, as rodent and cellular models poorly recapitulate the disease as seen in aging humans. To address this, we generated induced neurons (iNs) through direct reprogramming of human skin fibroblasts, which retain age-dependent epigenetic characteristics. HD-iNs displayed profound deficits in autophagy, characterised by reduced transport of late autophagic structures from the neurites to the soma. These neurite-specific alterations in autophagy resulted in shorter, thinner and fewer neurites specifically in HD-iNs. CRISPRi-mediated silencing of HTT did not rescue this phenotype but rather resulted in additional autophagy alterations in ctrl-iNs, highlighting the importance of wild type HTT in normal neuronal autophagy. In summary, our work identifies a distinct subcellular autophagy impairment in adult patient derived Huntington's disease neurons and provides a new rational for future development of autophagy activation therapies.
|
|
| 5. |
- Soust-Verdaguer, B., et al.
(author)
-
Implications of using systematic decomposition structures to organize building LCA information: A comparative analysis of national standards and guidelines- IEA EBC ANNEX 72
- 2020
-
In: IOP Conference Series: Earth and Environmental Science. - : IOP Publishing. - 1755-1307 .- 1755-1315. ; 588:2
-
Conference paper (peer-reviewed)abstract
- Introduction: The application of the Life Cycle Assessment (LCA) technique to a building requires the collection and organization of a large amount of data over its life cycle. The systematic decomposition method can be used to classify building components, elements and materials, overcome specific difficulties that are encountered when attempting to complete the life cycle inventory and increase the reliability and transparency of results. In this paper, which was developed in the context of the research project IEA EBC Annex 72, we demonstrate the implications of taking such approach and describe the results of a comparison among different national standards/guidelines that are used to conduct LCA for building decomposition. Methods: We initially identified the main characteristics of the standards/guidelines used by Annex participant countries. The “be2226” reference office building was used as a reference to apply the different national standards/guidelines related to building decomposition. It served as a basis of comparison, allowing us to identify the implications of using different systems/standards in the LCA practice, in terms of how these differences affect the LCI structures, LCA databases and the methods used to communicate results. We also analyzed the implications of integrating these standards/guidelines into Building Information Modelling (BIM) to support LCA. Results: Twelve national classification systems/ standards/guidelines for the building decomposition were compared. Differences were identified among the levels of decomposition and grouping principles, as well as the consequences of these differences that were related to the LCI organization. In addition, differences were observed among the LCA databases and the structures of the results. Conclusions: The findings of this study summarize and provide an overview of the most relevant aspects of using a standardized building decomposition structure to conduct LCA. Recommendations are formulated on the basis of these findings.
|
|
| 6. |
|
|
| 7. |
|
|
| 8. |
|
|
| 9. |
- Rasmussen, F. N., et al.
(author)
-
Existing benchmark systems for assessing global warming potential of buildings : Analysis of IEA EBC Annex 72 cases
- 2022
-
In: IOP Conference Series. - : IOP Publishing.
-
Conference paper (peer-reviewed)abstract
- Life cycle assessment (LCA) is increasingly being used as a tool by the building industry and actors to assess the global warming potential (GWP) of building activities. In several countries, life cycle based requirements on GWP are currently being incorporated into building regulations. After the establishment of general calculation rules for building LCA, a crucial next step is to evaluate the performance of the specific building design. For this, reference values or benchmarks are needed, but there are several approaches to defining these. This study presents an overview of existing benchmark systems documented in seventeen cases from the IEA EBC Annex 72 project on LCA of buildings. The study characterizes their different types of methodological background and displays the reported values. Full life cycle target values for residential and non-residential buildings are found around 10-20 kg CO2e/m2/y, whereas reference values are found between 20-80 kg CO2e/m2/y. Possible embodied target- and reference values are found between 1-12 kg CO2e/m2/y for both residential and non-residential buildings. Benchmark stakeholders can use the insights from this study to understand the justifications of the background methodological choices and to gain an overview of the level of GWP performance across benchmark systems.
|
|
| 10. |
- Costa, Jason, et al.
(author)
-
Combined assessment of sex- and mutation-specific information for risk stratification in type 1 long QT syndrome
- 2012
-
In: Heart Rhythm. - : Elsevier BV. - 1547-5271. ; 9:6, s. 892-898
-
Journal article (peer-reviewed)abstract
- BACKGROUND Men and women with type 1 long QT syndrome (LQT1) exhibit time-dependent differences in the risk for cardiac events. OBJECTIVE We hypothesized that sex-specific risk for LQT1 is related to the location and function of the disease-causing mutation in the KCNQ1 gene. METHODS The risk for life-threatening cardiac events (comprising aborted cardiac arrest [ACA] or sudden cardiac death [SCD]) from birth through age 40 years was assessed among 1051 individuals with LQT1 (450 men and 601 women) by the location and function of the LQT1-causing mutation (prespecified as mutations in the intracellular domains linking the membrane-spanning segments [ie, S2-S3 and S4-S5 cytoplasmic loops] involved in adrenergic channel regulation vs other mutations). RESULTS Multivariate analysis showed that during childhood (age group: 0-13 years) men had >2-fold (P < .003) increased risk for ACA/SCD than did women, whereas after the onset of adolescence the risk for ACA/SCD was similar between men and women (hazard ratio = 0.89 [P = .64]). The presence of cytoplasmic-loop mutations was associated with a 2.7-fold (P < .001) increased risk for ACA/SCD among women, but it did not affect the risk among men (hazard ratio 1.37; P = .26). Time-dependent syncope was associated with a more pronounced risk-increase among men than among women (hazard ratio 4.73 [P < .001] and 2.43 [P = .02], respectively), whereas a prolonged corrected QT interval (>= 500 ms) was associated with a higher risk among women than among men. CONCLUSION: Our findings suggest that the combined assessment of clinical and mutation location/functional data can be used to identify sex-specific risk factors for life-threatening events for patients with LQT1.
|
|
| 11. |
- Drouin-Ouellet, Janelle, et al.
(author)
-
REST suppression mediates neural conversion of adult human fibroblasts via microRNA-dependent and -independent pathways
- 2017
-
In: EMBO Molecular Medicine. - : EMBO. - 1757-4684 .- 1757-4676. ; 9:8, s. 1117-1131
-
Journal article (peer-reviewed)abstract
- Direct conversion of human fibroblasts into mature and functional neurons, termed induced neurons (iNs), was achieved for the first time 6 years ago. This technology offers a promising shortcut for obtaining patient- and disease-specific neurons for disease modeling, drug screening, and other biomedical applications. However, fibroblasts from adult donors do not reprogram as easily as fetal donors, and no current reprogramming approach is sufficiently efficient to allow the use of this technology using patient-derived material for large-scale applications. Here, we investigate the difference in reprogramming requirements between fetal and adult human fibroblasts and identify REST as a major reprogramming barrier in adult fibroblasts. Via functional experiments where we overexpress and knockdown the REST-controlled neuron-specific microRNAs miR-9 and miR-124, we show that the effect of REST inhibition is only partially mediated via microRNA up-regulation. Transcriptional analysis confirmed that REST knockdown activates an overlapping subset of neuronal genes as microRNA overexpression and also a distinct set of neuronal genes that are not activated via microRNA overexpression. Based on this, we developed an optimized one-step method to efficiently reprogram dermal fibroblasts from elderly individuals using a single-vector system and demonstrate that it is possible to obtain iNs of high yield and purity from aged individuals with a range of familial and sporadic neurodegenerative disorders including Parkinson's, Huntington's, as well as Alzheimer's disease.
|
|
| 12. |
- L'Episcopo, F, et al.
(author)
-
GSK-3β-induced Tau pathology drives hippocampal neuronal cell death in Huntington's disease : involvement of astrocyte-neuron interactions
- 2016
-
In: Cell Death and Disease. - : Springer Science and Business Media LLC. - 2041-4889. ; 7, s. 1-14
-
Journal article (peer-reviewed)abstract
- Glycogen synthase kinase-3β (GSK-3β) has emerged as a critical factor in several pathways involved in hippocampal neuronal maintenance and function. In Huntington's disease (HD), there are early hippocampal deficits both in patients and transgenic mouse models, which prompted us to investigate whether disease-specific changes in GSK-3β expression may underlie these abnormalities. Thirty-three postmortem hippocampal samples from HD patients (neuropathological grades 2-4) and age- and sex-matched normal control cases were analyzed using real-time quantitative reverse transcription PCRs (qPCRs) and immunohistochemistry. In vitro and in vivo studies looking at hippocampal pathology and GSK-3β were also undertaken in transgenic R6/2 and wild-type mice. We identified a disease and stage-dependent upregulation of GSK-3β mRNA and protein levels in the HD hippocampus, with the active isoform pGSK-3β-Tyr(216) being strongly expressed in dentate gyrus (DG) neurons and astrocytes at a time when phosphorylation of Tau at the AT8 epitope was also present in these same neurons. This upregulation of pGSK-3β-Tyr(216) was also found in the R6/2 hippocampus in vivo and linked to the increased vulnerability of primary hippocampal neurons in vitro. In addition, the increased expression of GSK-3β in the astrocytes of R6/2 mice appeared to be the main driver of Tau phosphorylation and caspase3 activation-induced neuronal death, at least in part via an exacerbated production of major proinflammatory mediators. This stage-dependent overactivation of GSK-3β in HD-affected hippocampal neurons and astrocytes therefore points to GSK-3β as being a critical factor in the pathological development of this condition. As such, therapeutic targeting of this pathway may help ameliorate neuronal dysfunction in HD.
|
|
| 13. |
- Migdalovich, Dimitry, et al.
(author)
-
Mutation and gender-specific risk in type 2 long QT syndrome: Implications for risk stratification for life-threatening cardiac events in patients with long QT syndrome
- 2011
-
In: Heart Rhythm. - : Elsevier BV. - 1547-5271. ; 8:10, s. 1537-1543
-
Journal article (peer-reviewed)abstract
- BACKGROUND Men and women with type 2 long QT syndrome (LQT2) exhibit time-dependent differences in the risk for cardiac events. We hypothesized that data regarding the location of the disease-causing mutation in the KCNH2 channel may affect gender-specific risk in LQT2. OBJECTIVE This study sought to risk-stratify LQT2 patients for life-threatening cardiac events based on clinical and genetic information. METHODS The risk for life-threatening cardiac events from birth through age 40 years (comprising aborted cardiac arrest [ACA] or sudden cardiac death [SCD]) was assessed among 1,166 LQT2 male (n = 490) and female (n = 676) patients by the location of the LQTS-causing mutation in the KCNH2 channel (prespecified in the primary analysis as pore-loop vs. non-pore-loop). RESULTS During follow-up, the cumulative probability of life-threatening cardiac events years was significantly higher among LQT2 women (26%) as compared with men (14%; P <.001). Multivariate analysis showed that the risk for life-threatening cardiac events was not significantly different between women with and without pore-loop mutations (hazard ratio 1.20; P = .33). In contrast, men with pore-loop mutations displayed a significant > 2-fold higher risk of a first ACA or SCD as compared with those with non-pore-loop mutations (hazard ratio 2.18; P = .01). Consistently, women experienced a high rate of life-threatening events regardless of mutation location (pore-loop: 35%, nonpore-loop: 23%), whereas in men the rate of ACA or SCD was high among those with pore-loop mutations (28%) and relatively low among those with non-pore-loop mutations (8%). CONCLUSION Combined assessment of clinical and mutation-specific data can be used for improved risk stratification for life-threatening cardiac events in LQT2.
|
|
| 14. |
- Bider, D, et al.
(author)
-
Guidelines for conducting epidemiological studies of blast injury
- 2019
-
In: Journal of the Royal Army Medical Corps. - : BMJ. - 0035-8665 .- 2052-0468. ; 165:1, s. 41-44
-
Journal article (peer-reviewed)abstract
- Blast injuries are often caused by more than one mechanism, do not occur in isolation, and typically elicit a secondary multi-system response. Research efforts often do not separate blast injuries caused by blast waves from those caused by blunt force trauma and other mechanisms. 15 experts from nine different NATO nations developed in the HFM Research Task Group (RTG; HFM-234 (RTG)) ‘Environmental Toxicology of Blast Exposures: Injury Metrics, Modelling, Methods and Standards’ Guidelines for Conducting Epidemiological Studies of Blast Injury. This paper describes these guidelines, which are intended to provide blast injury researchers and clinicians with a basic set of recommendations for blast injury epidemiological study design and data collection that need to be considered and described when conducting prospective longitudinal studies of blast injury.
|
|
| 15. |
- Birtele, Marcella, et al.
(author)
-
Dual modulation of neuron-specific microRNAs and the REST complex promotes functional maturation of human adult induced neurons
- 2019
-
In: FEBS Letters. - : Wiley. - 0014-5793 .- 1873-3468. ; 593:23, s. 3370-3380
-
Journal article (peer-reviewed)abstract
- Direct neuronal reprogramming can be achieved using different approaches: by expressing neuronal transcription factors or microRNAs; and by knocking down neuronal repressive elements. However, there still exists a high variability in terms of the quality and maturity of the induced neurons obtained, depending on the reprogramming strategy employed. Here, we evaluate different long-term culture conditions and study the effect of expressing the neuronal-specific microRNAs, miR124 and miR9/9*, while reprogramming with forced expression of the transcription factors Ascl1, Brn2, and knockdown of the neuronal repressor REST. We show that the addition of microRNAs supports neuronal maturation in terms of gene and protein expression, as well as in terms of electrophysiological properties.
|
|
| 16. |
- Bruzelius, Andreas, et al.
(author)
-
Reprogramming human adult fibroblasts into GABAergic interneurons
- 2021
-
In: Cells. - : MDPI AG. - 2073-4409. ; 10:12
-
Journal article (peer-reviewed)abstract
- Direct reprogramming is an appealing strategy to generate neurons from a somatic cell by forced expression of transcription factors. The generated neurons can be used for both cell replacement strategies and disease modelling. Using this technique, previous studies have shown that γ-aminobutyric acid (GABA) expressing interneurons can be generated from different cell sources, such as glia cells or fetal fibroblasts. Nevertheless, the generation of neurons from adult human fibroblasts, an easily accessible cell source to obtain patient-derived neurons, has proved to be challenging due to the intrinsic blockade of neuronal commitment. In this paper, we used an optimized protocol for adult skin fibroblast reprogramming based on RE1 Silencing Transcription Factor (REST) inhibitn together with a combination of GABAergic fate determinants to convert human adult skin fibroblasts into GABAergic neurons. Our results show a successful conversion in 25 days with upregulation of neuronal gene and protein expression levels. Moreover, we identified specific gene combinations that converted fibroblasts into neurons of a GABAergic intraneuronal fate. Despite the well-known difficulty in converting adult fibroblasts into functional neurons in vitro, we could detect functional maturation in the induced neurons. GABAergic interneurons have relevance for cognitive impairments and brain disorders, such as Alzheimer’s and Parkinson’s diseases, epilepsy, schizophrenia and autism spectrum disorders.
|
|
| 17. |
- Cisbani, G, et al.
(author)
-
Cystamine/cysteamine rescues the dopaminergic system and shows neurorestorative properties in an animal model of Parkinson's disease.
- 2015
-
In: Neurobiology of Disease. - : Elsevier BV. - 0969-9961. ; 82, s. 430-444
-
Journal article (peer-reviewed)abstract
- The neuroprotective properties of cystamine identified in pre-clinical studies have fast-tracked this compound to clinical trials in Huntington's disease, showing tolerability and benefits on motor symptoms. We tested whether cystamine could have such properties in a Parkinson's disease murine model and now provide evidence that it can not only prevent the neurodegenerative process but also can reverse motor impairments created by a 6-hydroxydopamine lesion 3weeks post-surgery. Importantly, we report that cystamine has neurorestorative properties 5weeks post-lesion as seen on the number of nigral dopaminergic neurons which is comparable with treatments of cysteamine, the reduced form of cystamine used in the clinic, as well as rasagiline, increasingly prescribed in early parkinsonism. All three compounds induced neurite arborization of the remaining dopaminergic cells which was further confirmed in ex vivo dopaminergic explants derived from Pitx3-GFP mice. The disease-modifying effects displayed by cystamine/cysteamine would encourage clinical testing.
|
|
| 18. |
- Collins, Lucy M., et al.
(author)
-
Dermal fibroblasts from patients with Parkinson's disease have normal GCase activity and autophagy compared to patients with PD and GBA mutations
- 2018
-
In: F1000Research. - : F1000 Research Ltd. - 2046-1402. ; 6
-
Journal article (peer-reviewed)abstract
- Background: Recently, the development of Parkinson's disease (PD) has been linked to a number of genetic risk factors, of which the most common is glucocerebrosidase (GBA) mutations. Methods: We investigated PD and Gaucher Disease (GD) patient derived skin fibroblasts using biochemistry assays. Results: PD patient derived skin fibroblasts have normal glucocerebrosidase (GCase) activity, whilst patients with PD and GBA mutations have a selective deficit in GCase enzyme activity and impaired autophagic flux. Conclusions: This data suggests that only PD patients with a GBA mutation have altered GCase activity and autophagy, which may explain their more rapid clinical progression.
|
|
| 19. |
|
|
| 20. |
- Drouin-Ouellet, Janelle, et al.
(author)
-
Age-related pathological impairments in directly reprogrammed dopaminergic neurons derived from patients with idiopathic Parkinson's disease
- 2022
-
In: Stem Cell Reports. - : Elsevier BV. - 2213-6711. ; 17:10, s. 2203-2219
-
Journal article (peer-reviewed)abstract
- We have developed an efficient approach to generate functional induced dopaminergic (DA) neurons from adult human dermal fibroblasts. When performing DA neuronal conversion of patient fibroblasts with idiopathic Parkinson's disease (PD), we could specifically detect disease-relevant pathology in these cells. We show that the patient-derived neurons maintain age-related properties of the donor and exhibit lower basal chaperone-mediated autophagy compared with healthy donors. Furthermore, stress-induced autophagy resulted in an age-dependent accumulation of macroautophagic structures. Finally, we show that these impairments in patient-derived DA neurons leads to an accumulation of phosphorylated alpha-synuclein, the classical hallmark of PD pathology. This pathological phenotype is absent in neurons generated from induced pluripotent stem cells from the same patients. Taken together, our results show that direct neural reprogramming can be used for obtaining patient-derived DA neurons, which uniquely function as a cellular model to study age-related pathology relevant to idiopathic PD.
|
|
| 21. |
- Drouin-Ouellet, Janelle, et al.
(author)
-
Direct neuronal reprogramming for disease modeling studies using patient-derived neurons : What have we learned?
- 2017
-
In: Frontiers in Neuroscience. - : Frontiers Media SA. - 1662-4548 .- 1662-453X. ; 11:SEP
-
Research review (peer-reviewed)abstract
- Direct neuronal reprogramming, by which a neuron is formed via direct conversion from a somatic cell without going through a pluripotent intermediate stage, allows for the possibility of generating patient-derived neurons. A unique feature of these so-called induced neurons (iNs) is the potential to maintain aging and epigenetic signatures of the donor, which is critical given that many diseases of the CNS are age related. Here, we review the published literature on the work that has been undertaken using iNs to model human brain disorders. Furthermore, as disease-modeling studies using this direct neuronal reprogramming approach are becoming more widely adopted, it is important to assess the criteria that are used to characterize the iNs, especially in relation to the extent to which they are mature adult neurons. In particular: i) what constitutes an iN cell, ii) which stages of conversion offer the earliest/optimal time to assess features that are specific to neurons and/or a disorder and iii) whether generating subtype-specific iNs is critical to the disease-related features that iNs express. Finally, we discuss the range of potential biomedical applications that can be explored using patient-specific models of neurological disorders with iNs, and the challenges that will need to be overcome in order to realize these applications.
|
|
| 22. |
|
|
| 23. |
|
|
| 24. |
- Merlevede, Adriaan, et al.
(author)
-
A quantitative model of cellular decision making in direct neuronal reprogramming
- 2021
-
In: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 11:1
-
Journal article (peer-reviewed)abstract
- The direct reprogramming of adult skin fibroblasts to neurons is thought to be controlled by a small set of interacting gene regulators. Here, we investigate how the interaction dynamics between these regulating factors coordinate cellular decision making in direct neuronal reprogramming. We put forward a quantitative model of the governing gene regulatory system, supported by measurements of mRNA expression. We found that nPTB needs to feed back into the direct neural conversion network most likely via PTB in order to accurately capture quantitative gene interaction dynamics and correctly predict the outcome of various overexpression and knockdown experiments. This was experimentally validated by nPTB knockdown leading to successful neural conversion. We also proposed a novel analytical technique to dissect system behaviour and reveal the influence of individual factors on resulting gene expression. Overall, we demonstrate that computational analysis is a powerful tool for understanding the mechanisms of direct (neuronal) reprogramming, paving the way for future models that can help improve cell conversion strategies.
|
|
| 25. |
- Shrigley, Shelby, et al.
(author)
-
Simple Generation of a High Yield Culture of Induced Neurons from Human Adult Skin Fibroblasts
- 2018
-
In: Journal of Visualized Experiments. - : MyJove Corporation. - 1940-087X. ; 132
-
Journal article (peer-reviewed)abstract
- Induced neurons (iNs), the product of somatic cells directly converted to neurons, are a way to obtain patient-derived neurons from tissue thatis easily accessible. Through this route, mature neurons can be obtained in a matter of a few weeks. Here, we describe a straightforward andrapid one-step protocol to obtain iNs from dermal fibroblasts obtained through biopsy samples from adult human donors. We explain each stepof the process, including the maintenance of the dermal fibroblasts, the freezing procedure to build a stock of the cell line, seeding of the cellsfor reprogramming, as well as the culture conditions during the conversion process. In addition, we describe the preparation of glass coverslipsfor electrophysiological recordings, long-term coating conditions, and fluorescence activated cell sorting (FACS). We also illustrate examplesof the results to be expected. The protocol described here is easy to perform and can be applied to human fibroblasts derived from human skinbiopsies from patients with various different diagnoses and ages. This protocol generates a sufficient amount of iNs which can be used for a widearray of biomedical applications, including disease modeling, drug screening, and target validation.
|
|
