№ files_lp_4_process_3_115184
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Supplementary experimental methods section of a biomedical research study detailing laboratory procedures for culturing glioblastoma-related cells, isolating blood exosomes, engineering Exos-Met/sicPLA2 complexes, and analyzing cellular uptake, membrane dynamics, and mitochondrial activity.
Document Type: Supplementary Methods
Scientific Field: Molecular Biology; Cancer Biology; Nanomedicine
Research Focus: Exosome-mediated delivery of metformin and siRNA in glioblastoma cells
Biological Materials: Primary GBM cells (N9, TBD0220); murine brain endothelial cells (bEnd.3); GBM cell lines (GL261, CT2A); neuronal cells (HT22); astrocyte cells (C8-DIA)
Key Molecules: PTRF protein; cPLA2; metformin; siRNA (siPTRF, chol-sicPLA2)
Experimental Techniques: Cell culture; lentiviral transfection; siRNA transfection; ultracentrifugation; western blot analysis; confocal laser scanning microscopy; flow cytometry; FRAP membrane fluidity assay; electroporation; nanoparticle tracking analysis; transmission electron microscopy; dynamic light scattering; high-performance liquid chromatography; Seahorse extracellular flux analysis
Biological Structures Studied: Blood-derived exosomes
Analytical Software: ImageJ
Laboratory Equipment: CLSM (FV1200 Olympus); NanoSight NS300; JEM-2100F TEM; Bio-Rad Gene Pulser Xcell Electroporation System; Seahorse XF24 Analyzer
Chemical Reagents: fetal bovine serum; metformin; DiI dye; Cy5 dye; oligomycin; FCCP; rotenone; antimycin A
Purpose of Experiments: Investigation of cellular uptake, membrane fluidity, mitochondrial function, and delivery efficiency of engineered exosomes in GBM cells
Geographical Reference of Reagents: Shanghai, China; Suzhou, China; Beijing, China; USA
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Urine exosome mRNA-based test for monitoring kidney allograft rejection: Effects of sample transportation and storage, and interference substances
Year: 2023
Region / city: West Hills, CA, United States
Topic: Kidney allograft rejection monitoring
Document type: Research article
Organization / institution: Thermo Fisher Scientific
Authors: McFaul M, Ventura C, Evans S, Dundar H, Rumpler MJ, McCloskey C, Lowe D, Vlassov AV
Target audience: Researchers, clinicians in organ transplant and diagnostics
Period of validity: 2023
Date of approval: October 23, 2023
Date of changes: None
Keywords: Kidney allograft, Post-transplant monitoring, Liquid biopsy, Exosome, mRNA, Quantitative polymerase chain reaction
Corresponding author: Alexandre V Vlassov, PhD
Summary: Research on the stability of mRNA from urine-derived exosomes for kidney allograft rejection detection, including effects of sample transportation, storage, and interference substances.
Glioblastoma Multiforme: References
Note: Year
Author: Willie Ching, PT, DPT, Board Certified Specialist in Neurologic Physical Therapy
Contextual Description: A fact sheet referencing various studies and research articles on Glioblastoma Multiforme, including treatment options and clinical trials.
Glioblastoma Multiforme: Background and Epidemiology/Statistics
Author: Willie Ching, PT, DPT, Board Certified Specialist in Neurologic Physical Therapy
Organization: American Physical Therapy Association (APTA) – Neurology Section
Document Type: Fact Sheet
Medical Condition: Glioblastoma Multiforme (GBM)
Target Audience: Healthcare professionals, patients, caregivers
Diagnostic Methods: MRI, biopsy
Treatment Methods: Surgery, radiation therapy, chemotherapy (Temozolomide), tumor-treating fields (TTFields)
Epidemiology: Incidence 2–3 per 100,000 adults per year, median age 64, higher prevalence in men and Caucasians
Survival Rates: 26–33% at 2 years, 4–5% at 5 years
Risk Factors: Ionizing radiation, certain chemical exposures, genetic syndromes (neurofibromatosis, tuberous sclerosis, Li-Fraumeni, retinoblastoma, Turcot syndrome)
Contact: ANPT, Phone 952.646.2038, [email protected]
Note: , www.neuropt.org
Glioblastoma Multiforme: Role of Physical Therapy and Research Updates
Year: 2023
Region / City: United States
Topic: Glioblastoma Multiforme, Physical Therapy, Research Updates
Document Type: Fact Sheet
Organization: ANPT
Author: Willie Ching, PT, DPT, Board Certified Specialist in Neurologic Physical Therapy
Target Audience: Healthcare professionals, patients with GBM, caregivers
Period of Action: Ongoing
Approval Date: Not specified
Date of Changes: Not specified
M2 Muscarinic Receptor Stimulation Induces Autophagy in Human Glioblastoma Cancer Stem Cells via mTOR Complex-1 Inhibition
Year: 2021
Region / City: Rome, Italy
Field: Cancer Research
Document Type: Article
Organization / Institution: Sapienza University of Rome, University of Milan, Consiglio Nazionale delle Ricerche (CNR), University of Trento, University of Ferrara
Authors: Guerriero, Claudia; Manfredelli, Marianna; Matera, Carlo; Iuzzolino, Angela; Conti, Luciano; Dallanoce, Clelia; De Amici, Marco; Trisciuoglio, Daniela; Tata, Ada Maria
Target Audience: Researchers in Cancer Biology
Period of Validity: N/A
Approval Date: N/A
Modification Date: N/A
Keywords: glioblastoma; cancer stem cells; M2 muscarinic receptor; orthosteric and dualsteric muscarinic agonism; autophagy; apoptosis; mTORC1
Funding: Ateneo Sapienza Funds
References: Aoki H, 2008, Autophagy, V4, P467, DOI 10.4161/auto.5668
Context: Scientific article exploring the role of M2 muscarinic receptor stimulation in inducing autophagy and apoptosis in glioblastoma cancer stem cells, and the differential effects of orthosteric and dualsteric agonists.
Supplementary Materials and Methods for ICOSLG-mediated IL-10–Producing Regulatory T Cell Expansion Promotes Progression of Glioblastoma Multiforme
Document Type: Supplementary research methods
Research Field: Tumor immunology
Disease Focus: Glioblastoma multiforme
Molecular Focus: ICOSLG-mediated signaling and IL-10–producing regulatory T cells
Experimental Models: Human glioma stem cells, murine RasR2 glioma-initiating cells, CD4+ naïve T cells
Biological Materials: Patient-derived neurospheres, peripheral blood mononuclear cells
Techniques: Flow cytometry, immunofluorescence staining, luciferase reporter assay, siRNA transfection, hypoxia induction, cytometry by time-of-flight (CyTOF)
Cell Culture Conditions: Defined DMEM/F12 medium with B27, heparin, EGF, and bFGF
Key Molecular Targets: ICOSLG, NF-κB, Foxp3, STAT5, PD-L1, HLA molecules
Reagents and Antibodies: Commercial reagents and antibodies from multiple biotechnology suppliers
Associated Experiments: Analysis of ICOSLG expression, NF-κB promoter activity, tumor-conditioned medium effects on T cells
Source Material: Surgical glioma specimens and established experimental cell lines
REST-DRD2 mechanism impacts glioblastoma stem cell-mediated tumorigenesis
Year: 2017
Institution: The University of Texas MD Anderson Cancer Center
Region / City: Houston, TX, USA
Document type: Supplementary materials
Subject: Glioblastoma stem cells, REST and DRD2 pathways
Methods: Cell culture, immunofluorescence, RNA preparation, qRT-PCR, chromatin immunoprecipitation, genome-wide expression analysis, Western blot, ChIP-Seq
Target audience: Researchers in molecular biology and oncology
Approval: Institutional review board approved
Experimental models: HR-GSC1, HR-GSC2, LR-GSC cell lines
Key reagents: Lentiviral vectors, antibodies, Trizol, microarrays
Analysis software: Agilent GeneSpring GX, Ingenuity Pathway Analysis
Supplementary content: Materials and methods, figure legends
Topics: Exos-MetsicPLA2 exosome Glioblastoma