Cell line construction and maintenance for Lyso-IP with or without genes linked with lysosomal storage disease
By onAssociated with the following preprint (published September 30th 2021): Progranulin deficiency results in reduced bis(monoacylglycero)phosphate (BMP) levels and gangliosidosis Sebastian Boland, Sharan Swarup, Yohannes A. Ambaw, Ruth C. Richards, Alexander W. Fischer, Shubham Singh, Geetika Aggarwal, Salvatore Spina, Alissa L. Nana, Lea T. Grinberg, William W. Seeley, Michal A. Surma, Christian Klose, Joao A. Paulo, Andrew D. Nguyen, J. Wade Harper, Tobias C. Walther, Robert V. Farese Jr. bioRxiv 2021.09.30.461806; doi: https://doi.org/10.1101/2021.09.30.461806
Freezing of hPSCs grown on MEFs
By onThis protocol describes the standard procedure of freezing human pluripotent stem cells (hPSCs), which were grown on inactivated mouse embryonic fibroblasts (MEFs).
Custom open-chamber microfluidic fabrication
By onThis protocol described the fabrication of two- and three-open chambered microfluidics suitable for cell culturing using commercially acquired master mould.
Cell-based Tau seeding assay
By onProtocol outlines Tau seeding techniques in cellular model for research purposes.
Human neuroblastoma cell line SH-SY5Y culturing
By onThis is the brief protocol for seeding and culturing human neuroblastoma SH-SY5Y cells.
Mitomycin C inactivation of mouse embryonic fibroblasts (MEFs) for hPSC cultures
By onThis protocol describes the process of using Mitomycin C to inactivate mouse embryonic fibroblasts (MEFs), which can then be used as feeder cells for human pluripotent stem cell (hPSC) culture.
Cell culture on EM grids and fluorescence microscopy imaging
By onThis protocol describes the general procedure of seeding mammalian cells on EM grids and confocal fluorescence microscopy imaging of grids for subsequent cryo-tomography experiments, performed.
Generation of stable cell lines via retroviral or lentiviral transduction
By onThis protocol details how to generate stable cell lines using a retrovirus system
Whole-cell patch-clamping of cultured human neurons
By onThis protocol describes procedures of the whole-cell patch clamping of human neurons derived from induced pluripotent stem cells and cultured in adherent monolayer.
Duplicating 96-well plate-cultured hPSCs clones
By onThis protocol describes a standard procedure for duplicating 96-well plate-cultured human pluripotent stem cells (hPSCs).
Primary neuronal cultures
By onThis protocol is associated with the following preprint, published on February 19th 2022: The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds Itika Saha, Patricia Yuste-Checa, Miguel Da Silva Padilha, Qiang Guo, Roman Körner, Hauke Holthusen, Victoria A. Trinkaus, Irina Dudanova, Rubén Fernández-Busnadiego, Wolfgang Baumeister, David W. Sanders, Saurabh Gautam, Marc I. Diamond, F. Ulrich Hartl, Mark S. Hipp bioRxiv 2022.02.18.481043; doi: https://doi.org/10.1101/2022.02.18.481043
Thawing of hPSCs grown on MEFs
By onThis protocol describes the standard procedure of thawing human pluripotent stem cells (hPSCs) on inactivated mouse embryonic fibroblasts (MEFs).
Transfection and protein over-expression
By onThis protocol provides the details for how to transfect and harvest 3X-FLAG-SHIP164Δ901-1099
TBK1 knockdown and rescue in Hela-M cells
By onThis protocol transiently depletes TBK1 from HeLa cells and re-introduces a tagged TBK1 along with other relevant components of mitophagy.
Protocol for hippocampal neuronal cultures
By onThis protocol details the procedue for preparation of neuronal culturs from mice hippocampi as it was performed in Guillén-Samander et al. 2021 article "VPS13D bridges the ER to mitochondria and peroxisomes via Miro" but can also be used to prepare culturs of cortical neurons.
Protocol Collection for “PARK15/FBXO7 is dispensable for PINK1/Parkin mitophagy in iNeurons and HeLa cell systems”
By onCollection of protocols associated with preprint (https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1.full) Protocols: Microscopy-based evaluation of Parkin translocation and mitophagy in FBXO7-/- cell lines Microscopy-based pUb-coverage measurements of mitochondria in iNeurons Microscopy-based measurements of p62 recruitment in HeLa Microscopy-based mtDNA turnover measurements in HeLa and iNeurons Microscopy-based mitochondrial morphology measurements in iNeurons
Preparing MEF-cultured hPSCs for nucleofection
By onThis protocol describes the procedure or preparing MEF-cultured human pluripotent stem cells (hPSCs) for the delivery of plasmids, mRNA, or ribonucleoprotein (RNP) using nucleofection.
